Aims. Orthopaedic infection is a potentially serious complication of elective and emergency trauma and orthopaedic procedures, with a high associated burden of morbidity and cost. Optimization of vitamin D levels has been postulated to be beneficial in the prevention of orthopaedic infection. This study explores the role of vitamin D in orthopaedic infection through a systematic review of available evidence. Methods. A comprehensive search was conducted on databases including Medline and Embase, as well as grey literature such as Google Scholar and The World Health Organization Database. Pooled analysis with weighted means was undertaken. Results. Pooled analysis of four studies including 651 patients found the mean 25(OH)D level to be 50.7 nmol/l with a mean incidence of infection of 70%. There was a paucity of literature exploring prophylactic 25(OH)D supplementation on reducing orthopaedic infection, however, there was evidence of association between low 25(OH)D levels and increased incidence of orthopaedic infection. Conclusion. The results indicate a significant proportion of orthopaedic patients have low 25(OH]D levels, as well as an association between low 25(OH)D levels and orthopaedic infection, but more randomized controlled trials need to be conducted to establish the benefit of prophylactic supplementation and the optimum regimen by dose and time. Cite this article: Bone Jt Open 2021;2(9):721–727

Aim. This study aimed to evaluate the impact on length of hospital stay from dedicated infectious diseases input for orthopaedic infection patients compared to sporadic infection specialist input. Method. We conducted an observational cohort study of 157 adults with orthopaedic infections at a teaching hospital in the UK. The orthopaedic infections included were: osteomyelitis, septic arthritis, infected metalwork and prosthetic joint infections, and adults were aged 18 years or more. Prior to August 2016, advice on orthopaedic infection patients was adhoc with input principally from the on-call infectious diseases registrar and phone calls to microbiology whereas after August 2016 these patients received regular input from dedicated infectious diseases doctor(s). The dedicated input involved bedside reviews, medical management, correct antimicrobial prescribing, managing adverse drug reactions, increased use of outpatient parenteral antimicrobial therapy (OPAT) services especially self-administration of intravenous antibiotics and shared decision-making for treatment failure, whilst remaining under orthopaedic team care. Orthopaedic patients operated on for management of their infection between 29/8/16 and 15/3/17 were prospectively identified and orthopaedic operation records were used to retrospectively identified patients between 29/8/15 and 15/3/16. The length of stay was compared between the 2 groups. Results. There were 83 patients in the dedicated infectious diseases input group (dedicated group) and 74 patients in the sporadic infection specialist input group (sporadic group). The dedicated group were significantly younger: median 58 years versus 69years (p<0.001), and there was a trend to significant differences in the breakdown of diagnosis (p=0.06), but no significant sex difference. The median length of stay for the sporadic group was 20 days (interquartile range (IQR) 13–29 days) compared to 14 days (IQR 9–27 days) for the dedicated group, with a trend to significance (p=0.06) but no effect from age or diagnosis. Our hospital values one day in hospital at £864, therefore over the 6.5 months trial period of the dedicated infectious diseases input there was a cost saving of £430,272 (£864 × 6 days × 83 patients). Conclusions. Dedicated infectious diseases input would be expected to improve patient care but by additionally reducing median length of stay for orthopaedic infection patients, this encourages investment to achieve both. In this era of increased scrutiny of health budgets demonstrating value for money, not just improved quality of patient care, is essential

Aim. Diagnosing Orthopaedic infection is limited by the sensitivity of culture methods. Next generation sequencing (NGS) offers an alternative approach for detection of microorganisms from clinical specimens. However, the low ratio of pathogen DNA to human DNA often inhibits detection of microorganisms from specimens. Depletion of human DNA may enhance the detection of microbial DNA. 1. Our aim was to compare four DNA extraction methods for the recovery of microbial DNA from orthopaedic samples for NGS. Method. Simulated samples; pooled culture negative sample matrix was spiked with known concentrations of microorganisms, each panel consisting of 7 samples. Broth culture was performed on simulated samples for comparison with NGS. *. . DNA Extraction; total nucleic acid extraction was performed on an automated extraction platform. **. using the viral NA assay. Modifications included: (1) mechanical lysis (glass beads), (2) lysis of human cells (saponin 0.025%), turbo DNase treatment and (3) mechanical lysis and addition of MspJI enzyme post-extraction for methylated DNA digestion. Detection of human and microbial DNA; human endogenous (HE) gene rtPCR. ***. was utilised following manufacturer's recommendations. Microbial DNA was detected using SYBR green 16s ribosomal RNA rtPCR with high resolution melt-curve analysis. ****. . Results. Broth culture recovered 64% (9/14) of the microorganisms from simulated samples. A significant increase (p<0.01) in the cycle threshold (C. T. ) (median C. T. 25.9 IQR 25.5, 26.1) of the HE gene rtPCR was observed using extraction method b, indicating a significant reduction in human DNA. No significant change (p=0.38) in the C. T. of the HE gene rtPCR was observed between the baseline method (median C. T. 19.2 IQR 18.5, 19.7) and modifications a (median C. T. 18.4 IQR 18.2, 19.4) and c (median C. T. 19.3 IQR 18.6, 19.4). Detection of microbial DNA was successful using the base line extraction method and modification a. Microbial DNA was not detected using the 16s ribosomal RNA rtPCR for modifications b and c. Conclusions. This study has demonstrated that modification of DNA extraction methods using selective enzymatic digestion of human DNA negatively impacts on the recovery of microbial DNA from simulated specimens. Total DNA extraction allows the successful recovery of microbial DNA alongside a significant amount of human DNA. The effect of the presence of human DNA will be subsequently assessed through NGS CosmosID analysis to establish if NGS is more sensitive than broth based culture

Local antimicrobial therapy is an integral aspect of treating orthopaedic device related infection (ODRI), which is conventionally administered via polymethylmethacrylate (PMMA) bone cement. PMMA, however, is limited by a suboptimal antibiotic release profile and a lack of biodegradability.

In this study, we compare the efficacy of PMMA versus an antibioticloaded hydrogel in a single- stage revision for chronic methicillin-resistant Staphylococcus aureus (MRSA) ODRI in

sheep. Antibiofilm activity of the antibiotic combination (gentamicin and vancomycin) was determined in vitro. Swiss alpine sheep underwent a single-stage revision of a tibial intramedullary nail with MRSA infection. Local gentamicin and vancomycin therapy was delivered via hydrogel or PMMA (n = 5 per group), in conjunction with systemic antibiotic therapy. In vivo observations included: local antibiotic tissue concentration, renal and liver function tests, and quantitative microbiology on tissues and hardware post-mortem.

There was a nonsignificant reduction in biofilm with an increasing antibiotic concentration in vitro (p = 0.12), confirming the antibiotic tolerance of the MRSA biofilm. In the in vivo study, four out of five sheep from each treatment group were culture negative. Antibiotic delivery via hydrogel resulted in 10–100 times greater local concentrations for the first 2–3 days compared with PMMA and were comparable thereafter. Systemic concentrations of gentamicin were minimal or undetectable in both groups, while renal and liver function tests were within normal limits.

This study shows that a single-stage revision with hydrogel or PMMA is equally effective, although the hydrogel offers certain practical benefits over PMMA, which make it an attractive proposition for clinical use.

Introduction and Aim: In 1995, sterile maggots (blow fly larvae) became available commercially for the first time since the mid-1930s. We have used them in managing ‘problem wounds’ in an orthopaedic unit. We have re-assessed the value of maggot debridement therapy (MDT) in present-day orthopaedics.

Method and Results: To date 95 patients have been treated. (Average age 62; range 16–91). Eighty-five percent of cases involved the lower limb. The remainder were upper limb, apart from one spinal lesion and one sacral sore. Twenty percent of patients had diabetes; six amputation stumps were treated. In 60% of cases a single application was used, the larvae being left in-situ for three to five days. Some wounds required up to three applications. The dressing technique is easily learnt and is ideal for outpatient clinics. The most appropriate wounds are those with a wide opening, extensive slough, and natural drainage. The greatest benefit follows infection with gram-positive cocci, and anaerobes. In eight cases, MRSA infection was cured or controlled.

Larvae provide optimal wound healing conditions, by literally eating pus and bacteria, and also by stimulating granulation tissue to form. However, they cannot produce wound healing if a major sequestrum or implant is present. In general, patient acceptance was good, but five patients requested early removal of maggots. Since 2001, the maggots have been available in sachet form (the so-called ‘Bio-bag’) and this packaged application has made the treatment more readily acceptable, and easier.

Overall we judged that MDT had produced healing or improvement in 80% of infected wounds. Unusual wounds, such as animal bites, a sea -urchin lesion, and infected gout produced some of the most striking cures.

Conclusion: Maggot therapy uniquely minimises both the need for surgical debridement and antibiotics. We therefore recommend its continued use.

Antimicrobial resistance (AMR) is projected to result in 10 million deaths every year globally by 2050. Without urgent action, routine orthopaedic operations could become high risk and musculoskeletal infections incurable in a “post-antibiotic era.” However, current methods of studying AMR processes including bacterial biofilm formation are 2D in nature, and therefore unable to recapitulate the 3D processes within in vivo infection.

Within this study, 3D printing was applied for the first time alongside a custom-developed bioink to bioprint 3D bacterial biofilm constructs from clinically relevant species including Staphylococcus aureus (MSSA), Methicillin-resistant staphylococcus aureus (MRSA), Escherichia coli and Pseudomonas aeruginosa. Bacterial viability and biofilm formation in bioprinted constructs was excellent, with confocal laser scanning microscopy (CSLM) used to demonstrate biofilm production and maturation over 28 days. Bioprinted 3D MRSA and MSSA biofilm constructs had greater resistance to antimicrobials than corresponding two-dimensional (2D) cultures. Thicker 3D E.coli biofilms had greater resistance to tetracycline than thinner constructs over 7 days of treatment. Raman spectroscopy was also adapted in a novel approach to non-invasively diagnose 3D bioprinted biofilm constructs located within a joint replacement model.

In conclusion, mature bacterial biofilm constructs were reproducibly 3D bioprinted for the first time using clinically relevant bacteria. This methodology allows the study of antimicrobial biofilm penetration in 3D, and potentially aids future antimicrobial research, replicating joint infection more closely than current 2D culture models. Furthermore, by deploying Raman spectroscopy in a novel fashion, it was possible to diagnose 3D bioprinted biofilm infections within a joint replacement model.

Introduction and Aims: A phenotypic and proteomic approach has identified novel targets for the development of a DNA vaccine to prevent Staphylococcus aureus infection in orthopaedics. Approximately 1% of joint replacement operations are complicated by infection. Thirty percent of these infections are due to S.aureus, which is often difficult to treat because of antibiotic resistance. As treatment of these infections is challenging, prevention with a vaccine is a very attractive option. Method: To infect a joint replacement, bacteria must first adhere to its surface. This adherence is mediated by specific adhesion proteins; the expression of which is controlled by virulence regulator genes within the bacterial cell. A DNA vaccine is being developed which targets this regulatory apparatus, thus preventing bacterial adhesion, allowing the immune system to rapidly clear any potential S.aureus infection. Results: Mutations of the agr,sar and sae virulence regulator genes have been made. Their properties have been explored using a flow cell system, which uses a scanning confocal laser microscope and image analysis software to accurately provide quantitative data in real-time of biofilm formation. We have shown that the sae mutant does not form biofilm in the same was as wild-type S.aureus. We have also shown that it does not adhere to steel as well as its wild-type counterpart. Conclusion: For such a dramatic difference in biofilm forming properties to be evident, there must be a difference in the adhesion proteins produced by the wild-type and the mutant bacteria. Gel-electrophoresis has compared protein expression of sae mutant and wild-type bacteria and identified differences. Those proteins which are not expressed in the non-biofilm-forming mutant are sequenced and from the protein sequences, DNA sequences are identified that will form part of the candidate DNA vaccine

Background

Staphylococcus epidermidis is one of the main organisms associated with prosthetic joint infections. One of the major pathogenic attributes of this organism is the ability to form biofilms, making it extremely resistant to currently available antimicrobial therapies. There is, therefore, an urgent requirement for novel agents that are effective against this organism. Antimicrobial peptides represent a novel group of agents that show good activity towards biofilm-forming S. epidermidis. Antimicrobial peptides are particularly interesting due to their multiple modes of action which are thought to reduce the rate of resistance development to the agents.

Orthopedic device-related infection (ODRI) preclinical models are widely used in translational research. Most models require induction of general anesthesia, which frequently results in hypothermia in rodents. This study aimed to evaluate the impact of peri anesthetic hypothermia in rodents on outcomes in preclinical orthopedic device-related infection studies. A retrospective analysis of all rodents that underwent surgery under general anesthesia to induce an ODRI model with inoculation of Staphylococcus epidermidis between 2016 and 2020 was conducted. A one-way multivariate analysis of covariance was used to determine the fixed effect of peri anesthetic hypothermia (hypothermic defined as rectal temperature <35°C) on the combined harvested tissue and implant colonies forming unit counts, and having controlled for the study groups including treatments received duration of surgery and anesthesia and study period. All animal experiments were approved by relevant ethical committee. A total of 127 rodents (102 rats and 25 mice) were enrolled in an ODRI and met the inclusion criteria. The mean lowest peri-anesthetic temperature was 35.3 ± 1.5 °C. The overall incidence of peri-anesthetic hypothermia was 41% and was less frequently reported in rats (34% in rats versus 68% in mice). Statistical analysis showed a significant effect of peri anesthetic hypothermia on the post-mortem combined colonies forming unit counts from the harvested tissue and implant(s) (p=0.01) when comparing normo- versus hypothermic rodents. Using Wilks’ Λ as a criterion to determine the contribution of independent variables to the model, peri-anesthetic hypothermia was the most significant, though still a weak predictor, of increased harvested colonies forming unit counts. Altogether, the data corroborate the concept that bacterial colonization is affected by abnormal body temperature during general anesthesia at the time of bacterial inoculation in rodents, which needs to be taken into consideration to decrease infection data variability and improve experimental reproducibility

One of the most common bacteria in orthopaedic prosthetic infections is Staphylococcus Aureus. Infection causes implant failure due to biofilm production. Biofilms are produced by bacteria once they have adhered to a surface. Nanotopography has major effects on cell behaviour. Our research focuses on bacterial adhesion on nanofabricated materials. We hypothesise that surface nanotopography impacts the differential ability of staphylococci species to adhere via altered metabolomics and may reduce orthopaedic implant infection rate. Bacteria were grown and growth conditions optimised. Polystyrene and titanium (Ti) nanosurfaces were studied. The polystyrene surfaces had different nanopit arrays, while the Ti surfaces expressed different nanowire structures. Adhesion analysis was performed using fluorescence imaging, quantitative PCR and bacterial percentage coverage calculations. Further substitution with ‘heavy’ labelled glucose into growth medium allowed for bacterial metabolomic analysis and identification of any up-regulated metabolites and pathways. Our data demonstrates reduced bacterial adhesion on specific nanopit polystyrene arrays, while nanowired titanium showed increased bacterial adhesion following qPCR (P<0.05) and percentage coverage calculations (P<0.001). Further metabolomic analysis identified significantly increased intensity counts of specific metabolites (Pyruvate, Aspartate, Alanine and Carbamoyl aspartate). Our study shows that by altering nanotopography, bacterial adhesion and therefore biofilm formation can be affected. Specific nanopatterned surfaces may reduce implant infection associated morbidity and mortality. The identification of metabolic pathways involved in adhesion may allow for a targeted approach to biofilm eradication in S. aureus. This is of significant benefit to both the patient and the surgeon, and may well extend far beyond the realms of orthopaedics

The most common bacteria in orthopaedic prosthetic infections are Staphylococcus, namely Staphylococcus Epidermidis (SE) and Staphylococcus Aureus (SA). Infection causes implant failure due to biofilm production. Biofilms are produced by bacteria once they have adhered to a surface. Nanotopography has major effects on cell behaviour. Our research focuses on bacterial adhesion and biofilm formation on nanofabricated materials. Bacteria studied were clinically relevant from an orthopaedic perspective, SA and SE. We hypothesise that that nanosurfaces can modulate bacterial adherence and biofilm formation and may reduce orthopaedic implant infection rate. Isolated bacteria were grown and growth conditions optimised. Bacterial concentrations were calculated by using qPCR. Statistical analysis allowed identification of optimal biofilm growth conditions. These were refined on standard, non-nanopatterned surfaces, and then control and nanopatterned polystyrene (nanopits) and titanium plates (nanowires). Adhesion analysis was performed using fluorescence imaging and quantitative PCR. 4 bacterial strains were isolated and cultured. Growth kinetics based on 24hr cultures allowed isolation of optimal media for biofilm conditions (Dulbecco's Modified Eagle Medium with additional supplements). Highest bacterial concentrations were found following 2hrs incubation with Lysozyme during qPCR. Bacterial concentration significantly increased between 30, 60 and 90 minutes incubation. Differences in percentage coverage on different polysyrene nanosurfaces (nanopits) were noted varying. This was confirmed by qPCR extractions that showed different bacterial concentrations on different nanopatterns. Titanium nanowire surfaces significantly increased bacterial adhesion (P<0.05). Our study cultured and quantified bacterial biofilm and suggests that by altering nanotopography, bacterial adhesion and therefore biofilm formation can be affected. Specific nanopatterned surfaces may reduce implant infection associated morbidity and mortality. Clearly this is of significant benefit to the patient, the surgeon and the NHS, and may well extend far beyond the realms of orthopaedics

Aim. In this study we investigated the effects of non-steroidal anti-inflammatory drugs (NSAIDs) with different cyclooxygenase (COX) selectivity on orthopaedic device-related infections (ODRIs) in a rat model. Specifically, we aimed to measure the impact of NSAID therapy on bone changes, bacterial load, and cytokine levels after treatment with antibiotics. In addition, we compared the effects of long vs short-term celecoxib (a COX-2 inhibitor) treatment on the same outcomes. Method. Skeletally mature female Wistar rats were implanted with Staphylococcus epidermidis-contaminated polyetheretherketone (PEEK) screws (1.5 × 10. 6. CFU per screw) in the proximal right tibia and monitored for 7 days. All animals received subcutaneous antibiotics (rifampicin plus cefazolin) for two weeks from day 7 to 21. In phase I of the study, rats were randomly assigned to receive 28 days of oral treatment with acetylsalicylic acid, ibuprofen, celecoxib, or vehicle control. In phase II, an additional group received seven days of celecoxib treatment from day 0 to 7. After implantation, bone changes were monitored using in vivo micro-CT and histology. Quantitative bacteriology was performed at euthanasia. Plasma samples were collected to measure cytokine levels at four time points (day 0, 6, 20, and 28). Results. The combination of antibiotic therapy resulted in treatment success in 85.71% of cases, while the addition of long-term celecoxib treatment reduced it to 45.45%. Long-term celecoxib treatment significantly reduced bone loss (33.85% mean difference [95% CI 14.12–53.58], p=0.0004 on day 6 bone fraction) and periosteal reaction (0.2760 μm mean difference [95% CI 0.2073–0.3448], p<0.0001 on day 14 periosteal thickness) during the early post-infection period compared to the control group. Short-term celecoxib treatment showed similar radiological results, however, there was no significant reduction in treatment success in the celecoxib group (88.9%). No differences in the selected inflammatory markers were observed. Conclusion. Our findings highlight the potential benefits of short-term use of celecoxib in improving bone fraction during the early post-infection period without impairing the efficacy of antibiotic therapy. This study suggests that celecoxib may be a useful addition to the multimodal approach to pain management in orthopaedic device-related infections

Aim. Culture of multiple periprosthetic tissue samples is the current gold-standard for microbiological diagnosis of prosthetic joint infections (PJI). Additional diagnostic information may be obtained through sonication fluid culture of explants. These current techniques can have relatively low sensitivity, with prior antimicrobial therapy or infection by fastidious organisms particularly influencing culture results. Metagenomic sequencing has demonstrated potential as a tool for diagnosis of bacterial, viral and parasitic infections directly from clinical samples, without the need for an initial culture step. We assessed whether metagenomic sequencing of DNA extracts from sonication fluid can provide a sensitive tool for diagnosis of PJI compared to sonication fluid culture. Method. We compared metagenomic sequencing with standard aerobic and anaerobic culture in 97 sonication fluid samples from prosthetic joint and other orthopaedic device-related infections. Sonication fluids were filtered to remove whole human cells and tissue debris, then bacterial cells were mechanically lysed before DNA extraction. DNA was sequenced and sequencing reads were taxonomically classified using Kraken. Using 50 derivation samples, we determined optimal thresholds for the number and proportion of bacterial reads required to identify an infection and confirmed our findings in 47 independent validation samples. Results. A total of 131 sonication fluids were aerobically and anaerobically cultured and underwent metagenomic sequencing. From the first 72 sonication fluid samples sequenced 22 samples from six batches were excluded, as these samples and negative controls from the same batches showed similar contamination. The remaining 50 samples, the derivation set, were used to determine optimal sequence thresholds for identifying true infection. Of 59 subsequently sequenced validation samples, 12 from a single batch were excluded as the negative control was contaminated with Propionibacterium acnes, leaving 47 validation samples. Compared to sonication fluid culture, the species-level sensitivity of metagenomic sequencing was 61/69(88%,95%CI 77–94%)(derivation samples 35/38[92%,79–98%]; validation samples 26/31[84%,66–95%]), and genus-level sensitivity was 64/69(93%,84–98%). Species-level specificity, adjusting for plausible fastidious causes of infection, species found in concurrently obtained tissue samples, and prior antibiotics, was 85/97(88%,79–93%)(derivation 43/50[86%,73–94%], validation 42/47[89%,77–96%]). High levels of human DNA contamination were seen despite use of laboratory methods to remove it. Conclusions. We demonstrate as a proof of principle that metagenomic sequencing can provide accurate diagnostic information in PJI. Further depletion of human DNA will lead to improved genomic information on the cause of infection, strengthening the case for metagenomic sequencing as a diagnostic tool in PJI

Aim. Diagnostics of orthopedic implant infection remains challenging and often shows false negative or inadequate results. Several methods have been described to improve diagnostic methods but most of them are expensive (PCR) or not accessible for all hospitals (sonication). Aim of this study was to evaluate the results of incubation of orthopedic explants compared to biopsies and punction fluid using conventional microbiological methods. Method. In this prospective study, we included patients who received septic or aseptic orthopedic implant removal in a single University hospital between July and December 2018. A part of the explant as well as minimum 2 tissue biopsies or additional punction fluid were put in a bouillon and incubated for 11 days. Patient´s records with co-morbidities, use of antibiotics and demographic data were evaluated. The results were analyzed. The study was approved by the ethical committee. Results. 94 patients were included in this study (43 females, 51 males, mean age 54 years). We detected statistically significant more pathogens in the bouillon with explants compared to biopsies (p=0,0059). We found the same results with pedicle screws (n=11, p=0,039) and endoprosthesis (n=56, p=0,019). Patients after osteosynthesis (p=27) showed same results but statistically not significant (p=0,050). Use of antibiotics did not have influence on the diagnostic result as well as co-morbidities. In 38 patients (40,4%), additional bacteria could be detected in explant´s bouillon. Most common pathogens were Staph. aureus, E. faecalis, Staph. epidermidis and Micrococcus luteus, mixed infections could be found in 9%. Conclusions. In this study we could show that incubation of orthopedic implants has advantages in diagnostics of pathogens in infected endoprosthesis, osteosynthesis and spondylodesis. This method is simple compared to PCR or sonication and as cheap as incubation of tissue samples but in 40% of the cases, additional pathogens can be detected. We recommend to incubate removed screws, hip endoprosthetic heads or inlays in bouillon to optimize diagnostics and to detect all pathogens

Aim. Orthopedic implant related surgical site infection (SSI) is a severe complication which represents an important challenge concerning to its treatment. Therefore, gram-negative orthopedic infections have recently become a global concern. Method. Retrospective study through searching of the SCIH (infection control service) database, concerning to the year 2016 and 2017. Cases selected were those of implant placement clean surgeries (osteosynthesis or prosthetic placement) which evolved with SSI and Gram-negative bacterial growth in bone tissue or periprosthetic cultures. Results. During 2016 and 2017, 6150 clean surgeries with orthopedic implant placement were performed; 140 fulfilled SSI criteria (83 cases of open fracture reduction, 44 of hip arthroplasty, 13 of knee arthroplasty). Main agent of infections was Staphylococcus aureus (32,47%) mostly of them methicillin-sensitive (69,20%). However, Gram-negative bacteria were responsible for 64,95% of infections. (Klebsiella pneumoniae 12.8%; Acinetobacter baumannii and Enterobacter ssp 11.96%; Pseudomonas aeruginosa 9.40%) Among them, 100% Enterobacter ssp. were sensitive to carbapenems and 75% to ciprofloxacin. Klebsiella pneumoniae showed sensitivity to carbapenems in 85.7%, Pseudomonas aeruginosa showed sensitivity in 85.7% to carbapenems and 100% to ciprofloxacin. Acinetobacter baumannii showed the least favorable profile amongst Gram-negatives since only 12.5% of strains were sensitive to carbapenems, 28.6% to Ampicilin-sulbactam, 22.2% to ciprofloxacin, while showing 100% sensitivity to polymyxins. 14 patients in whom Acinetobacter baumannii was isolated were predominantly elderly (median 70 years), most of them have underlying/chronic diseases (71.42%) such as diabetes, arterial hypertension, alcoholism, smoking and heart failure. None presented sepsis related to this infection, but four of them died as result of hospitalization related complications (28,60% mortality rate). Among these deaths, 3 were related to total hip arthroplasty, and one to knee arthroplasty. One patient died as result of external causes. Among the survivors, five showed remission/cure. The follow up was lost in 4 patients. Conclusions. SSI caused by carbepenem-resistant Acinetobacter baumannii represents considerable impact on morbi-mortality in patients who undergo surgery with placement of orthopedic implants

We investigated the effects of non-steroidal anti-inflammatory drugs (NSAIDs) with different cyclooxygenase (COX) selectivity on orthopaedic device-related infections (ODRIs) in a rat model. We aimed to measure the impact of NSAID therapy on bone changes, bacterial load, and cytokine levels after treatment with antibiotics. We also compared the effects of long vs short-term celecoxib (a COX-2 inhibitor) treatment on the same outcomes. Skeletally mature female Wistar rats were implanted with Staphylococcus epidermidis- contaminated polyetheretherketone (PEEK) screws in the proximal right tibia and monitored for 7 days. All animals received subcutaneous antibiotics (rifampicin plus cefazolin) for two weeks from day 7 to 21. In phase I of the study, rats were randomly assigned to receive 28 days of oral treatment with acetylsalicylic acid, ibuprofen, celecoxib, or vehicle control. In phase II, an additional group received seven days of celecoxib treatment from day 0 to 7. Bone changes were monitored using in vivo micro-CT and histology. Quantitative bacteriology was performed at euthanasia. Plasma samples were collected to measure cytokine levels on days 0, 6, 20, and 28. Combination antibiotic therapy resulted in treatment success in 85.71% of cases, while the addition of long-term celecoxib treatment reduced it to 45.45%. Long-term celecoxib treatment significantly reduced bone loss (33.85% mean difference [95% CI 14.12–53.58], p=0.0004 on day 6 bone fraction) and periosteal reaction (0.2760 μm mean difference [95% CI 0.2073–0.3448], p<0.0001 on day 14 periosteal thickness) during early infection compared to the control group. Short- term celecoxib treatment showed similar radiological results without a reduction in treatment success (88.9%). No differences in the inflammatory markers were observed. Our findings highlight the potential benefits of short-term use of celecoxib in improving bone fraction during the early post-infection period without impairing the efficacy of antibiotic therapy

Aim. Non-steroidal anti-inflammatory drugs (NSAIDs) are a cornerstone of perioperative pain management in orthopedic trauma surgery, although concerns persist regarding the potential impact of these drugs on fracture healing. Furthermore, NSAIDs may also exert an influence on host immune defenses, which may also be important in the context of infection treatment. However, this has been very much under-investigated in the clinical and scientific literature. The aim of this study was to determine the impact of NSAIDs on the course of an orthopedic device-related infection (ODRI) and its response to antibiotic therapy in a rat model. Method. A polyetheretherketone (PEEK) screw was inserted in the proximal tibia of 48 skeletally mature female Wistar rats: 12 control animals received a sterile screw, of which 6 also received NSAID therapy (carprofen, 5 mg/kg s.c. once daily); 36 rats received a Staphylococcus epidermidis-inoculated screw, of which 18 received NSAID therapy. Antibiotic therapy was administered from day 7–21 in 9 animals from all groups receiving S. epidermidis-inoculated screws (cefazolin: 30 mg/kg; s.c., b.i.d. plus rifampin: 25 mg/kg; s.c., b.i.d.). Bone histomorphometric changes were monitored using longitudinal microCT scanning, performed postoperatively, and at 3, 6, 9, 14, 20 and 28 days (euthanasia). Quantitative bacteriology of the implant, bone and overlying soft tissue was performed to assess infection status of individual animals. Results. All animals receiving S. epidermidis-inoculated screws in the absence of antibiotic therapy were confirmed as infected at euthanasia. Quantitative microbiology showed no significant change in bacterial load in NSAID-treated animals versus control. However, NSAID administration dramatically impaired antibiotic efficacy, with 7/8 animals remaining infected when NSAIDs were co-administered, whilst only 2/9 of control animals were infected when NSAIDs were withheld. Pronounced osteolysis was observed by day 6–9 in control animals, with reparative processes (periosteal proliferation and mineralization) observed at day 14. NSAID treatment markedly prevented S. epidermidis-induced osteolysis, but also reparative processes. Antibiotic treatment did not affect the bone changes. Conclusions. NSAID administration dramatically affected the response of bone tissue to infection, reducing osteolysis but also impairing reparative processes. Crucially, NSAIDs dramatically reduced antibiotic efficacy. Given these pronounced negative effects, further investigations should be conducted to determine the underlying pathophysiological mechanism and better understand the consequences of the therapeutic use of NSAIDs in human patients with ODRI

Collection of 4–5 independent peri-prosthetic tissue samples is recommended for microbiological diagnosis of prosthetic joint infections. Sonication of explanted prostheses has also been shown to increase microbiological yield in some centres. We compared sonication with standard tissue sampling for diagnosis of prosthetic joint and other orthopaedic device related infections. We used standard protocols for sample collection, tissue culture and sonication. Positive tissue culture was defined as isolation of a phenotypically indistinguishable organism from ≥2 samples; and positive sonication culture as isolation of an organism at ≥50 cfu/ml. We compared the diagnostic performance of each method against an established clinical definition of infection (Trampuz 2011), and against a composite clinical and microbiological definition of infection based on international consensus (Gehrke & Parvizi 2013). 350 specimens were received for sonication, including joint prostheses (160), exchangeable components (76), other orthopaedic hardware and cement (104), and bone (10). A median of 5 peri-prosthetic tissue samples were received from each procedure (IQR 4–5). Tissue culture was more sensitive than sonication for diagnosis of prosthetic joint and orthopaedic device related infection using both the clinical definition (66% versus 57%, McNemar's Χ2 test p=0.016) and the composite definition of infection (87% vs 66%, p<0.001). The combination of tissue culture and sonication provided optimum sensitivity: 73% (95% confidence interval 65–79%) against the clinical definition and 92% (86–96%) against the composite definition. Results were similar when analysis was confined to joint prostheses and exchangeable components; other orthopaedic hardware; and patients who had received antibiotics within 14 days prior to surgery. Tissue sampling appears to have higher sensitivity than sonication for diagnosis of prosthetic joint and orthopaedic device infection at our centre. This may reflect rigorous collection of multiple peri-prosthetic tissue samples. A combination of methods may offer optimal sensitivity, reflecting the anatomical and biological spectrum of prosthetic joint and other device related infections

Aims

Periprosthetic joint infections (PJIs) and fracture-related infections (FRIs) are associated with a significant risk of adverse events. However, there is a paucity of data on cardiac complications following revision surgery for PJI and FRI and how they impact overall mortality. Therefore, this study aimed to investigate the risk of perioperative myocardial injury (PMI) and mortality in this patient cohort.

Orthopedic device-related bone infection is one of the most distressing complications of the surgical fixation of fractures. Despite best practice in medical and surgical interventions, the rate of infection remains stubbornly persistent, and current estimates indicate that treatment failure rates are also significant. As we approach the limit of the effectiveness of current anti-infective preventative and therapeutic strategies, novel approaches to infection management assume great importance. This presentation will describe our efforts to develop and test various hydrogels to serve as customized antibiotic delivery vehicles for infection prevention and treatment. Hydrogels offer solutions for many of the challenges faced by complex trauma wounds as they are not restricted spatially within a poorly defined surgical field, they often degrade rapidly with no compatibility issues, and releases 100% of the loaded antibiotic. The preliminary data set proving efficacy in preventing and treating infection in both rabbit and sheep studies will be described, including local antibiotic concentrations in the intramedullary canal over time, compared to that of the more conventional antibiotic loaded bone cement. These two technologies show potential for the prevention and treatment of infection in trauma patients, with a clear focus on optimized antibiotic delivery tailored for complex wounds.

Objectives. Infection of implants is a major problem in elective and trauma surgery. Heating is an effective way to reduce the bacterial load in food preparation, and studies on hyperthermia treatment for cancer have shown that it is possible to heat metal objects with pulsed electromagnetic fields selectively (PEMF), also known as induction heating. We therefore set out to answer the following research question: is non-contact induction heating of metallic implants effective in reducing bacterial load in vitro?. Methods. Titanium alloy cylinders (Ti6Al4V) were exposed to PEMF from an induction heater with maximum 2000 watts at 27 kHz after being contaminated with five different types of micro-organisms: Staphylococcus epidermidis; Staphylococcus aureus; Pseudomonas aeruginosa; spore-forming Bacillus cereus; and yeast Candida albicans. The cylinders were exposed to incremental target temperatures (35°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C) for up to 3.5 minutes. Results. There was an average linear heating rate of 0.39°C per second up to the target temperature, and thereafter the target temperature was maintained until the end of the experiment. At 60°C and higher (duration 3.5 minutes), there was a 6-log reduction or higher for every micro-organism tested. At 60°C, we found that the shortest duration of effective induction heating was 1.5 minutes. This resulted in a 5-log reduction or higher for every micro-organism tested. Conclusion. Non-contact induction heating of a titanium disk is effective in reducing bacterial load in vitro. These promising results can be further explored as a new treatment modality for infections of metal orthopaedic implants. Cite this article: B. G. Pijls, I. M. J. G. Sanders, E. J. Kuijper, R. G. H. H. Nelissen. Non-contact electromagnetic induction heating for eradicating bacteria and yeasts on biomaterials and possible relevance to orthopaedic implant infections: In vitro findings. Bone Joint Res 2017;6:323–330. DOI: 10.1302/2046-3758.65.BJR-2016-0308.R1

Aims. Biofilm infections are among the most challenging complications in orthopaedics, as bacteria within the biofilms are protected from the host immune system and many antibiotics. Halicin exhibits broad-spectrum activity against many planktonic bacteria, and previous studies have demonstrated that halicin is also effective against Staphylococcus aureus biofilms grown on polystyrene or polypropylene substrates. However, the effectiveness of many antibiotics can be substantially altered depending on which orthopaedically relevant substrates the biofilms grow. This study, therefore, evaluated the activity of halicin against less mature and more mature S. aureus biofilms grown on titanium alloy, cobalt-chrome, ultra-high molecular weight polyethylene (UHMWPE), devitalized muscle, or devitalized bone. Methods. S. aureus-Xen36 biofilms were grown on the various substrates for 24 hours or seven days. Biofilms were incubated with various concentrations of halicin or vancomycin and then allowed to recover without antibiotics. Minimal biofilm eradication concentrations (MBECs) were defined by CFU counting and resazurin reduction assays, and were compared with the planktonic minimal inhibitory concentrations (MICs). Results. Halicin continued to exert significantly (p < 0.01) more antibacterial activity against biofilms grown on all tested orthopaedically relevant substrates than vancomycin, an antibiotic known to be affected by biofilm maturity. For example, halicin MBECs against both less mature and more mature biofilms were ten-fold to 40-fold higher than its MIC. In contrast, vancomycin MBECs against the less mature biofilms were 50-fold to 200-fold higher than its MIC, and 100-fold to 400-fold higher against the more mature biofilms. Conclusion. Halicin is a promising antibiotic that should be tested in animal models of orthopaedic infection. Cite this article: Bone Joint Res 2024;13(3):101–109

Abstract. Objectives. The objective of this study is to investigate if genomic sequencing is a useful method to diagnose orthopaedic infections. Current methods used to identify the species of bacteria causing orthopaedic infections take considerable time and the results are frequently insufficient for guiding antibiotic treatment. The aim here is to investigate if genomic sequencing is a faster and more reliable method to identify the species of bacteria causing infections. Current methods include a combination of biochemical markers and microbiological cultures which frequently produce false positive results and false negative results. Methods. Samples of prosthetic fluid were obtained from surgical interventions to treat orthopaedic infections. DNA is extracted from these samples lab and nanopore genomic sequencing is performed. Initial investigations informed that a sequencing time of 15 minutes was sufficient. The resulting genomic sequence data was classified using Basic Local Alignment Tool (BLAST) against the NCBI bacterial database and filtered by only including reads with an identity score of 90 and E-value of 1e-50. An E-value of 1e-50 suggests a high-quality result and is commonly used when analysing genomic data. This data was then filtered in R Studio to identify if any species were associated with orthopaedic infections. The results from genomic sequencing were compared to microbiology results from the hospital to see if the same species had been identified. The whole process from DNA extraction to output took approximately 2 hours, which was faster than parallel microbiological cultures. Results. In these preliminary analyses, 15 samples have been collected from patients with confirmed/suspected orthopaedic infections. To date, 11 samples from confirmed infected patients have been sequenced and a summary of the findings are presented in the table attached. As well as finding bacteria species to match microbiological cultures, genomic sequencing has also identified bacteria when culture results have been negative, but the patient is known to have an infection due to clinical indication and previous culture results. This example suggests genomic sequencing may have higher sensitivity than microbiological cultures at detecting bacteria causing orthopaedic infections. Results in table indicate the identification of bacteria from genomic sequencing that match microbiological cultures are high quality. Conclusions. Preliminary data presented using genomic sequencing suggests that the technique may be useful to identify bacterial species causing orthopaedic infections and can do so in a shorter time frame than current microbial methods. The results from genomic sequencing all produced a number of false positive results which hopefully can be reduced by improving the bioinformatic techniques used and increasing the sample number to include individuals without an infection. Further analysis will also look at identifying antibiotic resistance genes in the sequencing data and seeing if this ca be used to predict which patients will and will not respond to antibiotic treatment. The aim at the end of this project is to demonstrate if genomic sequencing is a more sensitive method to identify bacteria causing orthopaedic infections that current methods and if it can be used to guide antibiotic treatment. Include limitations, next steps and bigger picture. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

Aims

Biofilm-related infection is a major complication that occurs in orthopaedic surgery. Various treatments are available but efficacy to eradicate infections varies significantly. A systematic review was performed to evaluate therapeutic interventions combating biofilm-related infections on in vivo animal models.

Aim. Smoking is known to impair wound healing and to increase the risk of peri-operative adverse events and is associated with orthopaedic infection and fracture non-union. Understanding the magnitude of the causal effect on orthopaedic infection recurrence may improve pre-operative patient counselling. Methods. Four prospectively-collected datasets including 1173 participants treated in European centres between 2003 and 2021, followed up to 12 months after surgery for clinically diagnosed orthopaedic infections, were included in logistic regression modelling with Inverse Probability of Treatment Weighting for current smoking status [1–3]. Host factors including age, gender and ASA score were included as potential confounding variables, interacting through surgical treatment as a collider variable in a pre-specified structural causal model informed by clinical experience. The definition of infection recurrence was identical and ascertained separately from baseline factors in three contributing cohorts. A subset of 669 participants with positive histology, microbiology or a sinus at the time of surgery, were analysed separately. Results. Participants were 64% male, with a median age of 60 years (range 18–95); 16% of participants experienced treatment failure by 12 months. 1171 of 1173 participants had current smoking status recorded. As expected for the European population, current smoking was less frequent in older participants (Table 1). There was no baseline association between Charlson score or ASA score and smoking status (p=0.9, p=1, Chi squared test). The estimated adjusted odds ratio for treatment failure at 12 months, resulting from current smoking at the time of surgery, was 1.37 for all participants (95% CI 0.75 to 2.50) and 1.53 for participants with recorded confirmatory criteria (95% CI 1.14 to 6.37). Conclusions. Smoking contributes to infection recurrence, particularly in people with unequivocal evidence of osteomyelitis or PJI. People awaiting surgery for orthopaedic infection should be supported to cease smoking, not only to reduce anaesthetic risk, but to improve treatment outcomes. Limitations of this study include unmeasured socioeconomic confounding and social desirability bias resulting in uncertainty in true smoking status, resulting in underestimated effect size

Aim. Recurrence of bone and joint infection, despite appropriate therapy, is well recognised and stimulates ongoing interest in identifying host factors that predict infection recurrence. Clinical prediction models exist for those treated with DAIR, but to date no models with a low risk of bias predict orthopaedic infection recurrence for people with surgically excised infection and removed metalwork. The aims of this study were to construct and internally validate a risk prediction model for infection recurrence at 12 months, and to identify factors that predict recurrence. Predictive factors must be easy to check in pre-operative assessment and relevant across patient groups. Methods. Four prospectively collected datasets including 1173 participants treated in European centres between 2003 and 2021, followed up to 12 months after surgery for orthopaedic infections, were included in logistic regression modelling [1–3]. The definition of infection recurrence was identical and ascertained separately from baseline factors in three contributing cohorts. Eight predictive factors were investigated following a priori sample size calculation: age, gender, BMI, ASA score, the number of prior operations, immunosuppressive medication, glycosylated haemoglobin (HbA1c), and smoking. Missing data, including systematically missing predictors, were imputed using Multiple Imputation by Chained Equations. Weekly alcohol intake was not included in modelling due to low inter-observer reliability (mean reported intake 12 units per week, 95% CI for mean inter-rater error −16.0 to +15.4 units per week). Results. Participants were 64% male, with a median age of 60 years (range 18–95). 86% of participants had lower limb orthopaedic infections. 732 participants were treated for osteomyelitis, including FRI, and 432 for PJI. 16% of participants experienced treatment failure by 12 months. The full prediction model had moderate apparent discrimination: AUROC (C statistic) 0.67, Brier score 0.13, and reasonable apparent calibration. Of the predictors of interest, associations with failure were seen with prior operations at the same anatomical site (odds ratio for failure 1.51 for each additional prior surgery; 95% CI 1.02 to 2.22, p=0.06), and the current use of immunosuppressive medications (odds ratio for failure 2.94; 95% CI 0.89 to 9.77, p=0.08). Conclusions. This association between number of prior surgeries and treatment failure supports the urgent need to streamline referral pathways for people with orthopaedic infection to specialist multidisciplinary units

Revision surgeries for orthopaedic infections are done in two stages – one surgery to implant an antibiotic spacer to clear the infection and another to install a permanent implant. A permanent porous implant, that can be loaded with antibiotics and allow for single-stage revision surgery, will benefit patients and save healthcare resources. Gyroid structures can be constructed with high porosity, without stress concentrations that can develop in other period porous structures [1] [2]. The purpose of this research is to compare the resulting bone and prosthesis stress distributions when porous versus solid stems are implanted into three proximal humeri with varying bone densities, using finite element models (FEM). Porous humeral stems were constructed in a gyroid structure at porosities of 60%, 70%, and 80% using computer-aided design (CAD) software. These CAD models were analyzed using FEM (Abaqus) to look at the stress distributions within the proximal humerus and the stem components with loads and boundary conditions representing the arm actively maintained at 120˚ of flexion. The stem was assumed to be made of titanium (Ti6Al4V). Three different bone densities were investigated, representing a healthy, an osteopenic, and an osteoporotic humerus, with an average bone shape created using a statistical shape and density model (SSDM) based on 75 cadaveric shoulders (57 males and 18 females, 73 12 years) [3]. The Young's moduli (E) of the cortical and trabecular bones were defined on an element-by-element basis, with a minimum allowable E of 15 MPa. The Von Mises stress distributions in the bone and the stems were compared between different stem scenarios for each bone density model. A preliminary analysis shows an increase in stress values at the proximal-lateral region of the humerus when using the porous stems compared to the solid stem, which becomes more prominent as bone density decreases. With the exception of a few mesh dependent singularities, all three porous stems show stress distributions below the fatigue strength of Ti-6Al-4V (410 MPa) for this loading scenario when employed in the osteopenic and osteoporotic humeri [4]. The 80% porosity stem had a single strut exceeding the fatigue strength when employed in the healthy bone. The results of this study indicate that the more compliant nature of the porous stem geometries may allow for better load transmission through the proximal humeral bone, better matching the stress distributions of the intact bone and possibly mitigating stress-shielding effects. Importantly, this study also indicates that these porous stems have adequate strength for long-term use, as none were predicted to have catastrophic failure under the physiologically-relevant loads. Although these results are limited to a single boney geometry, it is based on the average shape of 75 shoulders and different bone densities are considered. Future work could leverage the shape model for probabilistic models that could explore the effect of stem porosity across a broader population. The development of these models are instrumental in determining if these structures are a viable solution to combatting orthopaedic implant infections

Introduction and Aims: The delivery of local antibiotics from a biodegradable implant for orthopedic infections is an attractive alternative. The implant delivers high tissue levels of antibiotic, obliterates dead space, aides in bone repair and does not require removal. This is a review of our clinical experience with custom-made calcium sulfate antibiotic implants (Osteoset BVF®, Wright Medical Technology, Arlington, TN). Method: Between 12/1996 and 1/2003, 156 procedures using biodegradable beads for orthopaedic infections were performed. Two patients were lost to follow-up. One elderly patient died from post-operative respiratory failure. One patient was lost to follow-up. There were 154 procedures in the analysis group performed on 145 patients (55 female: 90 male). The average age of the population was 45 years (1–85 years). The inclusion in the study required that a patient have an orthopedic infection (osteomyelitis/septic arthritis), which was either biopsy- or culture-proven. The population included 36 patients with a failed arthroplasty, 41 infected nonunions, 50 patients with chronic osteomyelitis, and 27 patients with acute or post-operative bone infections. Patients were treated initially with surgical debridement and insertion of calcium sulfate beads with either Tobramycin and/or Vancomycin. All patients received oral antibiotics or intravenous antibiotics. Results: Length of follow-up averaged 17 months (six to 67 months). Of the 154 patients, 142 became free of infection (92%). Five patients required multiple surgical procedures involving beads and further debridement to obtain an infection-free status. Many of these patients had an initial attempt at retaining a metal arthroplasty in an initial septic situation. In those patients, infection control required removal of the foreign material. Complete implant degradation occurred in all cases. Five patients had an episode post-operatively of early wound drainage, which was treated with sterile dressing changes while on intravenous antibiotics. In all patients the drainage ceased and the patients did not require further surgery. Conclusion: Biodegradable calcium sulfate antibiotic implants, along with adjunctive antibiotics, controlled a variety of infections, obliterated dead space and aided in bone repair. Subsequent bone grafting was not required for intramedullary defects due to the osteoconductive properties of calcium sulfate. Biodegradable antibiotic beads appear effective in controlling localised orthopedic infections and do not require a second procedure for removal

Correct diagnosis of infection is crucial for an adequate treatment of orthopedic implant-related infections. In the orthopedic field, infections can be difficult to diagnose(1). As a consequence, patients may suffer from an undiagnosed and untreated implant-related infection. To solve this problem, we are searching for a diagnostic method to detect these so-called low-grade infections. The technique fluorescence in situ hybridization (FISH) can detect slow-growing and even dead bacteria. Further, as FISH results are available within an hour after tissue collection it is an ideal candidate for diagnostic purposes. AIM: to evaluate the FISH technique for its potential to detect and identify orthopedic infections. Sonication fluid (SF) was collected by sonicating retrieved implants(2) from 62 patients. All samples were subjected to bacterial culture for clinical diagnostics. In addition, a commercially available FISH kit (miacom diagnostics, Germany), specifically designed for blood analysis (hemoFISH Masterpanel), was used. The kit contained 16S rRNA probes (positive control), non-sense probes (negative control), probes for Staphylococcus spp., Staphylococcus aureus, Streptococcus spp., Streptococcus pneumoniae, Streptococcus agalactiae, Enterococcus faecium, Enterococcus faecalis, Enterobacteriaceae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Acetinobacter spp., and Stenotrophomonas maltophilia. All FISH analyses were performed according to the protocol provided with the kit. Culture and FISH results were compared, considering culture as the gold standard. Culture resulted in 27 positive and 35 negative samples. Comparing FISH (16S rRNA probe) with culture, 24 samples tested true-positive and 32 samples true-negative. Furthermore, 3 samples tested false-negative and 3 samples false-positive. The species cultured with the highest incidence were Propionibacterium acnes and Staphylococcus epidermidis, both from 8 SF samples. As the kit did not contain a probe for Propionibacterium acnes, these strains were only detected by the 16S rRNA probe. In addition, the latter samples tested positive with the Staphylococcus spp. probe. Interestingly, 3 samples tested positive with FISH that were culture negative. This result could indicate a higher sensitivity for detection of bacteria with FISH than with culture. Before FISH can be used for diagnostic purposes, the technique needs to be optimized to prevent false-negative results, for use on other patient materials and for detection of bacterial strains relevant for the orthopedic field like Propionibacterium acnes. In conclusion, FISH holds promise to be used as a diagnostic tool for identifying orthopedic infections

Aims and Objectives. To assess the efficacy of linezolid in the treatment of orthopaedic related infection and the instance of adverse reactions. Methods. The management of 22 patients treated with oral linezolid for orthopaedic related infections were reviewed. Patients were selected from the hospital database using clinical coding related to orthopaedic infections and all patients were managed within a single tertiary referral centre. These included infected joint arthroplasty (10 patients), infection following fracture fixation (8 patients), septic arthritis and soft tissue infection (non trauma 4 patients). All patients were treated with oral linezolid therapy, and in each case treatment was initiated with the involvement of a microbiologist, as per trust anti-microbial policy. A diagnosis of infection was confirmed on basis of both subjective and objective markers. Results. 77% of patients resolution of infection at follow up (mean length of follow up 28 months) with mean C reactive protein showing a significant (p<0.05) reduction from 123 mg/l to 13.2 mg/l. This included 87.5% resolution in the infection following fracture fixation, 80% in the infected arthroplasty and 75% in the non trauma related infection groups. Adverse reactions were observed in 3 patients. Conclusion. This study demonstrates linezolid offers an excellent alternative treatment option, along with appropriate surgical management, for the management of orthopaedic related infections, with few adverse side effects. Traditionally, this cohort of patients has required lengthy in hospital admission for intra venous antibiotics. Our study demonstrates that use of linezolid may facilitate early hospital discharge in patients with orthopaedic related infections, since it is 100% bioavailable with oral administration, albeit with regular monitoring of blood parameters

Infection is one of the most serious complications of orthopedic surgery, particularly in implant-related procedures. Minimum inhibitory concentration (MIC) for identified bacteria is an important factor for successful antibiotic treatment. We investigated the MIC of antibiotics in Staphylococcus species from orthopedic infections, comparing with isolates from respiratory medicine. Staphylococcus species isolated in our laboratory from January 2013 to July 2016 were retrospectively reviewed. The MIC of vancomycin (VCM), arbekacin (ABK), teicoplanin (TEIC), linezolid (LZD), and rifampicin (RFP) was reviewed. Differences in the MIC of each antibiotic in orthopedic and respiratory samples were determined. A total of 259 isolates were evaluated (89 orthopedic, 170 respiratory). Staphylococcus aureus was the most commonly identified species (58%). In comparison with orthopedic samples, the number of isolates with a VCM MIC <0.5 μg/ml in methicillin sensitive staphylococcus aureus (MSSA) was significantly higher in respiratory isolates, while a MIC of 2 μg/ml was significantly lower (P = 0.0078). The proportion of isolates with a VCM MIC of 2 μg/ml in methicillin-resistant coagulase-negative staphylococci (MRCNS) was significantly higher in orthopedic isolates than that seen in respiratory isolates of methicillin-resistant staphylococcus aureus (MRSA; P < 0.001). When comparing MRCNS and other orthopedic Staphylococci, the rate of RFP MIC >2 μg/ml in MRCNS isolates was significantly higher (P = 0.0058). The MIC of VCM in Staphylococcus species from orthopedic infection was higher than that of respiratory samples, particularly in MRCNS from implant-related samples. MRCNS showed a significantly higher rate of resistance for RFP versus other orthopedic isolates

Antibiotic resistance represents a threat to human health. It has been suggested that by 2050, antibiotic-resistant infections could cause ten million deaths each year. In orthopaedics, many patients undergoing surgery suffer from complications resulting from implant-associated infection. In these circumstances secondary surgery is usually required and chronic and/or relapsing disease may ensue. The development of effective treatments for antibiotic-resistant infections is needed. Recent evidence shows that bacteriophage (phages; viruses that infect bacteria) therapy may represent a viable and successful solution. In this review, a brief description of bone and joint infection and the nature of bacteriophages is presented, as well as a summary of our current knowledge on the use of bacteriophages in the treatment of bacterial infections. We present contemporary published in vitro and in vivo data as well as data from clinical trials, as they relate to bone and joint infections. We discuss the potential use of bacteriophage therapy in orthopaedic infections. This area of research is beginning to reveal successful results, but mostly in nonorthopaedic fields. We believe that bacteriophage therapy has potential therapeutic value for implant-associated infections in orthopaedics. Cite this article: Bone Joint J 2021;103-B(2):234–244

The incidence of PJI in knee replacements is 2.8% and slightly lower with hip replacement surgery. PJI make up 15% (or even more) of knee revisions. To combat PJI, antibiotic laden bone cement has been used for many decades, but antibiotic stewardship dictates more prudent management of antimicrobials. Projected increase in infection rate, due to increased surgery and latent infection to be almost 5-fold up to 2035. Biofilm is a complex structure of bacteria and polysaccharide matrix and, is recognised as a major component in PJI and other orthopaedic infections. Biofilm is responsible for high incidence of resistance to antimicrobials and ineffective host immune response. Method. Stabilized hypochlorous acid has been reported to have a rapid kill rate on all pathogens, including MDR pathogens associated with chronic and acute wound infections. It destroys biofilm on contact, is not cytotoxic, reduces inflammation and stimulates wound healing. 0,038% of Hypochlorous acid was used as prophylaxis against infection and to treat PJI. We report on our experience with hypochlorous acid as a wound irrigation as prophylaxis against infection (more than 600 cases) and for PJI. We also report on a University study where a head to head analysis was done on the anti-biofilm efficacy between hypochlorous acid 0,038% (Trifectiv Surgical Wound Irrigation) and Product X (an industry-standard product for the prevention and treatment of biofilm infection. Hypochlorous acid offers a valuable addition to the armamentarium of wound antiseptics, with added anti-inflammatory value. An in vitro study demonstrated superior efficacy against biofilm when compared to Product X

Aims. Continuous local antibiotic perfusion (CLAP) has recently attracted attention as a new drug delivery system for orthopaedic infections. CLAP is a direct continuous infusion of high-concentration gentamicin (1,200 μg/ml) into the bone marrow. As it is a new system, its influence on the bone marrow is unknown. This study aimed to examine the effects of high-concentration antibiotics on human bone tissue-derived cells. Methods. Cells were isolated from the bone tissue grafts collected from six patients using the Reamer-Irrigator-Aspirator system, and exposed to different gentamicin concentrations. Live cells rate, apoptosis rate, alkaline phosphatase (ALP) activity, expression of osteoblast-related genes, mineralization potential, and restoration of cell viability and ALP activity were examined by in vitro studies. Results. The live cells rate (the ratio of total number of cells in the well plate to the absorbance-measured number of live cells) was significantly decreased at ≥ 500 μg/ml of gentamicin on day 14; apoptosis rate was significantly increased at ≥ 750 μg/ml, and ALP activity was significantly decreased at ≥ 750 μg/ml. Real-time reverse transcription-polymerase chain reaction results showed no significant decrease in the ALP and activating transcription factor 4 transcript levels at ≥ 1,000 μg/ml on day 7. Mineralization potential was significantly decreased at all concentrations. Restoration of cell viability was significantly decreased at 750 and 1,000 μg/ml on day 21 and at 500 μg/ml on day 28, and ALP activity was significantly decreased at 500 μg/ml on day 28. Conclusion. Our findings suggest that the exposure concentration and duration of antibiotic administration during CLAP could affect cell functions. However, further in vivo studies are needed to determine the optimal dose in a clinical setting. Cite this article: Bone Joint Res 2024;13(3):91–100

Aim. We reviewed a cohort of individuals with recurrent orthopaedic infection to describe the relative rates of microbial persistence vs re-infection at recurrence surgery. Method. A cohort of 125 individuals with recurrent infection (prosthetic joint infection, fracture-related infection and osteomyelitis) from two centres in the UK between 2007 and 2021. Electronic patient records were reviewed to identify culture results from surgical samples at index surgery and the next operation for recurrent infection. Antibiotic sensitivity results were recorded as sensitive, intermediate or resistant according to contemporary sensitivity testing guidelines. Results. Among patients with recurrent infection, 78/125 (62.4%) were male, with a median age 64 years (IQR 51–73y). 76 had prosthetic joint infection, and 49 had fracture related infection or osteomyelitis. Culture results at index procedure showed the most frequently isolated species were Staphylococci (Table 1). A single species was isolated in 75/125 (60%) and mixed species in 36/125 (28.8%). No organisms were cultured in 14/125 (11.2%). At re-operation 48/125 (38.4%) individuals had an organism from the same species or group as at the index operation. In 49/125 (39.2%), none of the organisms isolated at re-operation were grown at first operation. In 28/125 (22.4%), re-operative cultures yielded no growth. For each species isolated at the index procedure, the proportion with the same, different or no organisms isolated at the next procedure were reviewed (Table 1). Staphylococci (including S. aureus and coagulase-negative staphylococci) and Pseudomonas species showed the highest rate of persistence at the species level. Among coagulase-negative staphylococci, changes in antimicrobial sensitivity that make it unclear if these infections were truly persistent, or represented re-infection. Conclusions. Infection with different organisms was seen at similar rates (39.2% vs 38.4%) to persistent infection with the same species in this cohort. Staphylococcus aureus is the organism most likely to be persistently identified in recurrent infections

Aims. The aim of this study was to determine the diagnostic accuracy of α defensin (AD) lateral flow assay (LFA) and enzyme-linked immunosorbent assay (ELISA) tests for periprosthetic joint infection (PJI) in comparison to conventional synovial white blood cell (WBC) count and polymorphonuclear neutrophil percentage (PMN%) analysis. Methods. Patients undergoing joint aspiration for evaluation of pain after total knee arthroplasty (TKA) or total hip arthroplasty (THA) were considered for inclusion. Synovial fluids from 99 patients (25 THA and 74 TKA) were analyzed by WBC count and PMN% analysis, AD LFA, and AD ELISA. WBC and PMN% cutoffs of ≥ 1,700 cells/mm. 3. and ≥ 65% for TKA and ≥ 3,000 cells/mm. 3. and ≥ 80% for THA were used, respectively. A panel of three physicians, all with expertise in orthopaedic infections and who were blinded to the results of AD tests, independently reviewed patient data to diagnose subjects as with or without PJI. Consensus PJI classification was used as the reference standard to evaluate test performances. Results were compared using McNemar’s test and area under the receiver operating characteristic curve (AUC) analysis. Results. Expert consensus classified 18 arthroplasies as having failed due to PJI and 81 due to aseptic failure. Using these classifications, the calculated sensitivity and specificity of AD LFA was 83.3% (95% confidence interval (CI) 58.6 to 96.4) and 93.8% (95% CI 86.2 to 98.0), respectively. Sensitivity and specificity of AD ELISA was 83.3% (95% CI 58.6 to 96.4) and 96.3% (95% CI 89.6 to 99.2), respectively. There was no statistically significant difference between sensitivity (p = 1.000) or specificity (p = 0.157) of the two AD assays. AUC for AD LFA was 0.891. In comparison, AUC for synovial WBC count, PMN%, and the combination of the two values was 0.821 (sensitivity p = 1.000, specificity p < 0.001), 0.886 (sensitivity p = 0.317, specificity p = 0.011), and 0.926 (sensitivity p = 0.317, specificity p = 0.317), respectively. Conclusion. The diagnostic accuracy of synovial AD for PJI diagnosis is comparable and not statistically superior to that of synovial WBC count plus PMN% combined. Cite this article: Bone Joint J 2021;103-B(6):1119–1126

Aim. There is a lack of data supporting the use of doxycycline as a single agent after removing infected orthopaedic metalwork. We evaluated the efficacy and safety of doxycycline compared with other single antibiotic regimens used at our specialist orthopaedic hospital. Methods. A retrospective observational study including all adult patients diagnosed with an orthopaedic metalwork infection due to staphylococci. All patients were managed with the removal of metalwork, and multiple intraoperative samples were sent for culture, followed by the administration of at least four weeks of oral antibiotics. Antibiotic selection was on the recommendation of an infection consultant. Infection outcome was assessed as the proportion of patients meeting the OVIVA Trial definition of definite failure at follow-up. The probability of definite failure for doxycycline and the alternatives group was estimated using the Kaplan-Meier survival method. All adverse drug reactions (ADR) during treatment were analysed. Results. Seventy-nine orthopaedic metalwork infections were identified between July 2017 and July 2021. Forty-four were prosthetic joints, and 35 were fracture-related metalwork. In 54 cases, the infecting organism was Staphylococcus aureus, and 25 were due to coagulase-negative staphylococci. Forty-four were treated with doxycycline 100mg 12 hourly, and 35 were treated with alternatives (flucloxacillin 1g 6-hourly n=21 and clindamycin 450mg 6-hourly n=14). Overall, 70 patients (88.6%) were infection-free after a median follow-up of 23 months (IQR, 19 – 44). 38 (82.3%) were infection-free in the doxycycline group compared with 32 (91.4%) patients treated with alternatives. Of the failures in the alternatives group, all 3 received flucloxacillin. Survival analysis showed no significant difference in time to treatment failure between doxycycline and alternative antibiotics. Eighteen patients experienced an ADR: 2 nausea, one rash and one vaginal candidiasis due to doxycycline. Four diarrhoea, one reflux, two rashes and one headache due to clindamycin; 1 nausea and five diarrhoea due to flucloxacillin. Four patients required discontinuation therapy, two due to clindamycin and two due to flucloxacillin. Conclusions. In our cohort of patients, doxycycline monotherapy was an effective and well-tolerated oral option for treating staphylococcal infection following debridement and removal of orthopaedic metalwork

Aim. Bone and joint infection requires antimicrobial treatment for 6 to 12 weeks. When patients are well prepared and instructed regarding their therapy, they are more likely to have less side effects and improved compliance. Although side effects are common, this coaching is often not routinely performed when oral treatment is given. We developed a monitoring and guidance program for our outpatients who are on long term antimicrobial therapy, in which we can early signal side effects and treatment failure and coach the patients in their journey of infection treatment. Method. In our tertiary referral centre for orthopaedic infections, we started the outpatient monitoring of antimicrobial treatment (OMAT)- team for patients who will receive antimicrobial therapy for >2 weeks. Before discharge, our trained nurse gives instruction to the patient. Within 3 days after hospital discharge the patient is contacted by phone to, if necessary, clarify ambiguities in monitoring set up. During this contact, the nurse checks for side effects, addresses logistic problems regarding laboratory monitoring or future appointments and coaches patients for other questions. The patient is instructed how to recognize and who to contact in case of red flags and problems possibly related to the treatment. This is repeated after every laboratory check-up. Supervision is performed by an infectious disease specialist in close collaboration with the patient's surgeon. Results. The OMAT-team started in October 2020 and consists of 3 trained nurses and 3 ID specialist. In one year, 453 patients were proactively monitored for a mean of 11 weeks. Routinely, laboratory measurements were performed 1 week after the start of therapy and every 3–4 weeks thereafter, which resulted in 2711 contacts per year. In total, 64% of the patients reported side effects and 13% needed one or more extra laboratory measurement. This led to 40 additional outpatient consultations by the ID specialist because of complications of treatment and a switch of the antimicrobial agent in 31% of the patients. Conclusions. OMAT seems to improve the early signalling of complications regarding treatment, which is likely to improve compliance. The OMAT-team serves as a easy to access team to discuss any problem regarding antimicrobial therapy. Being proactive, the OMAT-team intervenes in an early stage of problems regarding side effects, logistics of the treatment and possible treatment failure. Future analysis of our data will show to what extend this will lead to prevention of re-hospitalization and improvement of success rate

Aim. Despite the expanding research focusing on bacterial biofilm formation, specific histochemical biofilm stains have not been developed for light microscopy. Therefore, pathologists are often not aware of the presence of biofilm formation when examining slides for diagnosing bacterial infections, including orthopaedic infections. The aim of the present study was to develop a combined histochemical and immunohistochemical biofilm stain for simultaneous visualization of Staphylococcus aureus bacteria and extracellular matrix in different colours using light microscopy. Methods. Infected bone tissue was collected from two different porcine models of osteomyelitis inoculated with the biofilm forming S. aureus strain S54F9. The infection time was 5 and 15 days, respectively. First, 25 common histochemical protocols were used in order to find stains that could identify extracellular biofilm matrix. Hereafter, the histochemical protocols for Alcian Blue pH3, Luna and Methyl-pyronin green were combined with an immunohistochemical protocol based on a specific antibody against S. aureus. Finally, the three new combined protocols were applied to infected bone tissue from a child suffering from chronic staphylococcal osteomyelitis for more than a year. For all combined protocols applied on all types of tissue (porcine and human) the number of double stained bacterial aggregates were counted. On the same sections the percentage of extracellular matrix of representative bacterial aggregates was calculated by image analysis. Results. Simultaneous visualization of bacterial cells and extracellular matrix in different colours was detected in both porcine and human tissue sections with all three combined protocols. The bacterial cells were red to light brown and the extracellular matrix either light blue, blue or orange depending on the histochemical stain i.e. if it was Alcian blue pH3 (colouring polysaccharides), Luna or Methyl green-pyronin (both colouring extracellular DNA), respectively. In the porcine models, 10 percent of the bacterial aggregates in a 10× magnification field revealed both the extracellular matrix and bacteria simultaneously in two different colours. For the human case, this was seen in 90 percent of the bacterial aggregates. The percentage of extracellular matrix of representative bacterial aggregates was 60 and 20 percent in the human and porcine tissues, respectively. Conclusions. The amount of S. aureus biofilm extracellular matrix increased with infection time. A combination of histochemical and immunohistochemical staining is a practical method for identification and evaluation of S. aureus biofilm in orthopaedic infections

Aim. The β-lactam penicillin is often used in the treatment of soft tissue infections and osteomyelitis caused by penicillin susceptible Staphylococcus aureus. Oral antibiotic treatment has been shown to be non-inferior to intravenous (IV) therapy when used during the first 6 weeks in complex orthopedic infections (OVIVA trial). However, the use of oral β-lactams in osteomyelitis treatment remains a topic of debate due to low and variable bioavailability. The aim was to assess the time for which the unbound penicillin concentration exceeded targeted minimum inhibitory concentrations (fT>MIC) in cancellous bone and subcutaneous tissue after IV (penicillin G) and oral (penicillin V) treatment in a porcine microdialysis model. Method. 12 female pigs (75kg) were assigned to standard clinical regimens of either three doses of IV penicillin G (1.2g) or oral penicillin V (0.8g) every 6h over 18h. Microdialysis catheters were placed for sampling in tibial cancellous bone and adjacent subcutaneous tissue. Data was collected in the first dosing interval (0–6h; prophylactic situation) and the third dosing interval (12–18h; assumed steady state). Plasma samples were collected for reference. MIC targets of 0.125μg/mL (Staph. aureus breakpoint), 0.25μg/mL (Strep. Group A, B, C and G breakpoint) and 0.5μg/mL (4xMIC) were applied. Results. For all investigated MIC targets, IV penicillin G resulted in a longer mean fT>MIC in cancellous bone during the first dosing interval, and in both cancellous bone and subcutaneous tissue during the third dosing interval compared to oral penicillin V. Across compartments, mean fT>MIC for IV penicillin G (MIC: 0.125, 0.25 and 0.5μg/mL) were ≥97%, ≥84% and ≥75% during the first dosing interval, and 100%, ≥95% and ≥88%, during the third dosing interval. The mean fT>MIC for oral penicillin V were ≥40%, ≥24% and ≥7% during the first dosing interval, and ≥42%, ≥36% and ≥18% during the third dosing interval. Conclusions. The findings suggest that standard clinical dosing of IV penicillin G provides superior fT>MIC in cancellous bone and subcutaneous tissue compared to oral penicillin V, particularly in the third dosing interval. This emphasizes the importance of appropriate route of administration when applying penicillin treatment. Acknowledgements. Funding was received from The Kirsten and Freddy Johansen Foundation, The Novo Nordisk Foundation, The Beckett Foundation, The Hede Nielsen Family Foundation, King Christian the 10. th. Foundation, The A.P. Møller Foundation, The Dagmar Marshalls Foundation, and The Carl and Ellen Hertz Foundation

Aim. Local antibiotic treatment for bone and joint infections offers direct delivery of high concentrations of antibiotics with reduced systemic exposure and favourable safety profile. However, the possibility of prolonged release of antibiotics at sub-therapeutic levels creates concern about the possible development of antimicrobial resistance. We investigated patients with recurrent bone and joint infection for evidence of antimicrobial resistance emerging from the use of local antibiotics. Method. 125 patients with recurrent infection (prosthetic joint infection, fracture related infection and osteomyelitis) in the UK between 2007 and 2021 were identified. Electronic patient records (including operative notes, pathology results and prescriptions) were reviewed to extract site of infection, date of surgery, the use of local antibiotics, culture results, empiric and definitive antibiotic therapy. All antibiotic sensitivity results were recorded as sensitive, intermediate or resistant according to contemporary guidelines (BSAC and EUCAST). Results. Local antibiotics were used in 74/125 (59.2%) of patients. Agents used were Gentamicin 53/125 (42.4%), Tobramycin 18/125 (14.4%), and vancomycin in 19/125 (15.2%). Combined gentamicin and vancomycin usage was seen in 16/125 patients (12.8%). Gentamicin non-sensitivity was common in this cohort with frequent aminoglycoside use. At index procedure, a Gentamicin non-sensitive organism was cultured in 51/125 patients (40.8%). At re-operation this proportion was lower: 40/125 (32%). There was no statistically significant difference in the rate of Gentamicin resistance at reoperation comparing patients who previously received local aminoglycosides with those who had not (21/71, 29.8% vs 19/54, 35.2% p=0.6, chi-squared test). In 48/125 (38.4%) of patients, the same species was isolated during the index and recurrence surgery. We identified 7 cases with new aminoglycoside resistance arising at the second procedure. In 2/7 – S. aureus and E. faecalis - aminoglycoside resistance was the only change in antimicrobial sensitivity. In 5/7, there were at least 2 additional changes in observed antimicrobial sensitivity. 3/74 (4%) of cases who initially received local aminoglycoside cultured organisms with aminoglycoside resistance at recurrence. 4/51 (7.8%) of those who did not receive local or systemic aminoglycoside at index surgery cultured resistant organisms (chi square 0.82; p=0.365). Conclusions. As a group, patients whose treatment for orthopaedic infection included local antibiotics did not exhibit higher rates of specific antimicrobial resistance compared with those not treated with local antibiotics. However we did identify cases where Gram positive bacteria developed aminoglycoside resistance regardless of their initial antimicrobial therapy. This should be considered in antimicrobial choice during surgery for recurrence

A growing number of recent investigations on the human genome, gut microbiome, and proteomics suggests that the loss of mucosal barrier function, particularly in the gastrointestinal tract, may substantially affect antigen trafficking, ultimately influencing the close bidirectional interaction between the gut microbiome and the immune system. This cross-talk is highly influential in shaping the host immune system and ultimately clinical infections. The hypothesis of the current study was that a change in microbiome and/or breach in GI epithelial barrier could be partially responsible for development of periprosthetic joint infections (PJI). Multiple biomarkers of gut barrier disruption were tested in parallel in plasma samples collected as part of a prospective cohort study of patients undergoing revision arthroplasty for aseptic failures or PJI (As defined by the 2018 ICM criteria). All blood samples were collected before any antibiotic was administered. Samples were tested for Zonulin, soluble CD14 (sCD14), and lipopolysaccharide (LPS) using commercially available enzyme-linked immunosorbent assays. Statistical analysis consisted of descriptive statistics, Mann-Whitney t-test, and Kruskal-Wallis test. A total of 134 patients were consented and included in the study. 44 were classified as PJI (30 chronic and 14 acute), and 90 as aseptic failures (26 primaries and 64 aseptic revisions). Both Zonulin and sCD14, but not LPS, were found to be significantly increased in the PJI group compared to non-infected cases (p<0.001; p=0.003). Higher levels of Zonulin were found in acute infections compared to chronic PJI (p=0.005. This prospective ongoing study reveals a possible link between gut permeability and the ‘gut-immune-joint axis’ in PJI. If this association continues to be born out with larger cohort recruitment and more in-depth analysis, it would have an immense implication in managing patients with PJI. In addition to administering antimicrobials, patients with PJI and other orthopedic infections may require gastrointestinal modulators such as pro and prebiotics

Aim. A growing number of recent investigations on the human genome, gut microbiome, and proteomics suggests that the loss of mucosal barrier function, particularly in the gastrointestinal tract, may substantially affect antigen trafficking, ultimately influencing the close bidirectional interaction between the gut microbiome and the immune system. This cross-talk is highly influential in shaping the host immune system function and ultimately shifting genetic predisposition to clinical outcome. Therefore, we hypothesized that a similar interaction could affect the occurrence of acute and chronic periprosthetic joint infections (PJI). Method. Multiple biomarkers of gut barrier disruption were tested in parallel in plasma samples collected as part of a prospective cohort study of patients undergoing revision arthroplasty for aseptic or PJI (As defined by the 2018 ICM criteria). All blood samples were collected before any antibiotic was administered. Samples were tested for Zonulin, soluble CD14 (sCD14), and lipopolysaccharide (LPS) using commercially available enzyme-linked immunosorbent assays. Statistical analysis consisted of descriptive statistics and ANOVA. Results. A total of 96 patients were consented and included in the study. 32 were classified as PJI (23 chronic and 9 acute), and 64 as aseptic. Both Zonulin and LPS were found to be increased in the acute PJI group 8.448 ± 7.726 ng/mL and 4.106 ± 4.260 u/mL, compared to chronic PJI (p<0.001) and aseptic revisions (p=0.025). sCD14 was found to be increased in both chronic (0.463 ± 0.168 ug/mL) and acute PJI (0.463 ± 0.389 ug/mL) compared to aseptic revisions (p<0.001). Conclusions. This prospective ongoing study reveals a possible link between gut permeability and the ‘gut-immune-joint axis’ in PJI. If this association continues to be born out with larger cohort recruitment, it would have a massive implication in managing patients with PJI. In addition to the administration of antimicrobials, patients with PJI and other orthopedic infections may require gastrointestinal modulators such as pro and prebiotics

Aim. Fungal orthopaedic infections most commonly affect people with complex surgical histories and existing comorbidities. Recurrence and re-infection rates are high, even with optimal surgical and systemic antifungal treatment. AmBisome liposomal amphotericin B has been suggested for local antifungal therapy, as an adjunctive treatment for fungal osteoarticular infections. Few case series have examined its clinical use when combined with polymethylmethacrylate cement PMMA), or with absorbable local antibiotic carriers. We aimed to evaluate the clinical use of local antifungal therapy with AmBisome liposomal amphotericin B (ABlaB), including tolerated doses, serious adverse events, and treatment outcomes. Method. A retrospective cohort of all patients treated with local antifungal therapy with ABlaB between January 2016 and January 2021 in a specialist orthopaedic hospital was identified using pharmacy records. Renal function, serious adverse events during treatment, surgical outcomes including spacer fracture and infection recurrence, were identified from electronic clinical records. The project was approved by the Institutional Review Board (clinical audit 6871). Results. 13 operations involving local antifungal therapy with ABlaB, in 12 patients, were identified. Eleven were infected with Candida species and one with Aspergillus. Mean follow-up was 22 months (range 4–46). Ten first stage arthroplasty revisions, 2 second stage arthroplasty revisions, and one debridement and removal of metalwork for fracture-related infection were performed. Locally implanted doses of ABlaB ranged from 100mg to 3600mg (50–400mg per 40g mix of PMMA). Six patients received ABlaB in absorbable antibiotic carriers containing calcium sulphate. This was noted to delay carrier setting. Patients were also given systemic antifungal therapy. No patients experienced serious adverse events related to toxicity from local antifungal therapy with ABlaB. There were no spacer fractures. Overall treatment success was 54% at final follow-up, although there were no recurrent fungal infections identified in patients experiencing treatment failure. Conclusions. Local antifungal therapy with liposomal amphotericin B, when combined with surgery and systemic therapy, appears to be a safe and well tolerated intervention in the management of complex fungal osteoarticular infections

Aims. There is a considerable challenge in treating bone infections and orthopaedic device-associated infection (ODAI), partly due to impaired penetration of systemically administrated antibiotics at the site of infection. This may be circumvented by local drug administration. Knowledge of the release kinetics from any carrier material is essential for proper application. Ceftriaxone shows a particular constant release from calcium sulphate (CaSO. 4. ) in vitro, and is particularly effective against streptococci and a large portion of Gram-negative bacteria. We present the clinical release kinetics of ceftriaxone-loaded CaSO. 4. applied locally to treat ODAI. Methods. A total of 30 operations with ceftriaxone-loaded CaSO. 4. had been performed in 28 patients. Ceftriaxone was applied as a single local antibiotic in 21 operations and combined with vancomycin in eight operations, and in an additional operation with vancomycin and amphotericin B. Sampling of wound fluid was performed from drains or aspirations. Ceftriaxone concentrations were measured by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Results. A total of 37 wound fluid concentrations from 16 operations performed in 14 patients were collected. The ceftriaxone concentrations remained approximately within a range of 100 to 200 mg/l up to three weeks. The median concentration was 108.9 mg/l (interquartile range 98.8 to 142.5) within the first ten days. No systemic adverse reactions were observed. Conclusion. Our study highlights new clinical data of locally administered ceftriaxone with CaSO. 4. as carrier material. The near-constant release of ceftriaxone from CaSO. 4. observed in vitro could be confirmed in vivo. The concentrations remained below known local toxicity thresholds. Cite this article: Bone Joint Res 2022;11(11):835–842

INTRODUCTION. Surgical site infections (SSI) in orthopaedics are a major source of postoperative morbidity. Although perioperative antibiotic prophylaxis is a common practice, orthopaedic infections are still high in numbers, due to the increasing use of osteosynthesis material and implants. Implants are avascular and can be easily colonized with biofilm-producing germs. For both, effective prophylaxis and treatment of orthopaedic infections, the right choice of the antibiotics used, the mode of application (only systemic or systemic & local), the timing, dosage and the duration of antibiotics are of extremely high importance. Their inappropriate use does not only lead to failures in prevention or treatment of infections, but may also promote microbial resistance development and may cause serious side effects for the patients. SELECTION & USE OF ANTIBIOTICS. Prophylaxis. Broad-spectrum prophylactic antibiotics should help to eliminate the germs before they start to colonize the implant. For prophylactic purposes the recently published AAOS guidelines [1] recommend the use of cephalosporins, such as cefazolin or cefuroxim, administered within one hour prior to surgery. In cases of suspected beta-lactam allergy, clindamycin or vancomycin can be used. The latter one is also recommended in cases of MRSA colonisation. Due to extended infusion times, vancomycin should be started within two hours prior to incision. In cases of blood loss or long op duration, antibiotic administration must be repeated (e.g. cefazolin, every 2–5 hrs; vancomycin, every 6–12 hrs). There is no evidence of a benefit of continued antibiotic administration past 24 hrs of end of surgery [2]. Treatment. In cases of established infections, use of antibiotics is only considered as an adjuvant to surgical debridement. Typically, the choice of the appropriate antibiotic depends on the bacteria, its antibiotic sensitivity profile and the health state of the patient. A combination of rifampicin & a quinolone (or rifampicin & vancomycin in cases of MRSA) for at least 2 wks up to several months has shown good results [3]. In chronic infections with biofilm involvement, all foreign material must be removed and locally delivered antibiotics via e.g. PMMA as carrier (spacers, PMMA-chains) are of additional clinical benefit. ROLE OF LOCAL ANTIBIOTICS. There is general consensus that PMMA chains or PMMA spacers loaded with specific antibiotics support the eradication of bone and joint infections, because of the high local concentrations achieved. The exact treatment time is, however, variable, ranging from few weeks up to several months. Only small amounts of these local antibiotics are systemically detectable and do not represent a major risk for side effects. Still a matter of debate is the benefit of antibiotic impregnated PMMA for infection prophylaxis. Although common practice in Europe, its routine use in e.g. primary arthroplasty is still discussed in other world regions. Meanwhile, evidence accumulates that joint infection rates are, indeed, lower, if antibiotic loaded bone cement with high initial release rates is routinely used in arthroplasty. 4.

Aim. To retrospectively investigate the clinical outcome after surgical, single-stage treatment of orthopaedic infections using antibiotics delivered locally by a calcium sulphate/hydroxyapatite biocomposite. Method. In order to identify the patients, we retrospectively searched several patient associated hospital-based databases using free text search with the term “Cerament” between November 2015 and November 2018. 58 cases with confirmed osteomyelitis and in which the bone substitute loaded with Gentamicin and/or Vancomycin had been used were identified and further evaluated. Results. Mean age was 58.9 years (range: 25–89). 46 (79.3 %) patients had at least 12 months follow up. The remaining 12 patients had a mean follow up time of 10.0 months (range 7–11). Infection was eradicated in 54 patients (93.1 %). In one the patients with recurrent infection repeated surgery with addition of bone substitute loaded with fosfomycin eventually eradicated the infection. One patient died of causes not related to the infection nor the treatment. Five patients presented transient white wound drainage but no signs of infection. No other side effects were identified. Conclusions. Local administration of antibiotics and dead space management using a calcium sulphate/hydroxyapatite biocomposite. 1. in combination with single-stage surgical debridement, stabilisation and postoperative culture-specific systemic antibiotics resulted in a high amount of eradicated infections and in line with other authors

Introduction The aim of this study was firstly to investigate the prevalence of icaABCD-operon which codes the production of the polysaccharide intracellular adhesin(PIA), responsible for biofilm production, in a collection of clinically significant staphylococci isolated from orthopaedic infections and secondly to assess the relationship between biofilm production and the presence or not of ica-operon. First Step – Material & Methods Between 1/2003 and 12/2005 200 CoNS were isolated from orthopaedic patients associated with soft tissue and bone infections(group I) and 200 CoNS from blood cultures of hospitalized patients from different wards of the same Hospital(group II). Identification was carried out by Gram-stain, catalase and coagulase tests and the API Staph System. Detection of icaADBC genes was performed by PCR. Production of biofilm was tested by the method of Christensen. Results In group I, 62(31.37%) carried the entire ica-operon; from these isolates biofilm formation was detected in 35(17.5%). 5 isolates, despite biofilm production, did not carry any gene of ica-operon. In group II, 70(35.5%) carried entire the ica-operon; biofilm formation was detected in 37(18.5%) of these isolates. 3 S. capitis, 1 S. epidermidis and 1 S. hominis carried only the icaADB, icaA and icaB genes respectively. Second Step – Material & Methods Based on the observation of PIA-production only in (50%) of ica(+) CoNS, 20 S. epidermidis isolates recovered from clinical specimens (pus) of orthopaedic patients and belonging to distinct PFGE clones, were selected on the basis of the presence of the entire ica operon. Nevertheless, only 10 of them produced biofilm. Nucleotide sequence analysis of ica-operon was carried out in all isolates; expression of icaADBC genes was also tested by RT-PCR. Results Sequencing analysis revealed that all isolates carried an intact ica-operon, without point mutations. Concerning icaADBC mRNA production, all genes of ica-operon were expressed in biofilm-producing isolates, whereas in the no-biofilm producing strains the icaA and icaC genes were not expressed, while a faint expression was observed for the icaB and icaD genes. Discussion Biofilm-forming capacities of CoNS from orthopaedic infections was not significantly greater than those from other infections (p> 0,05). The capacity of ica-operon(+) staphylococcal isolates to form biofilm seems to be dependent on the expression of ica-genes, specifically of icaA and icaC. The inability of ica(+) isolates to produce biofilm emphasizes that some unknown mechanisms influence icaADBC expression. Finally, the recognition of biofilm-producing CoNS without carrying any gene of ica operon underlined the existence of unidentified also mechanisms controlling biofilm production, apart from icaADBC expression

Methicillin-resistant Staphylococcus aureus (MRSA) has become a ubiquitous bacterium in both the hospital and community setting. There are two major subclassifications of MRSA, community-acquired and healthcare-acquired, each with differing pathogenicity and management. MRSA is increasingly responsible for infections in otherwise healthy, active adults. Local outbreaks affect both professional and amateur athletes and there is increasing public awareness of the issue. Health-acquired MRSA has major cost and outcome implications for patients and hospitals. The increasing prevalence and severity of MRSA means that the orthopaedic community should have a basic knowledge of the bacterium, its presentation and options for treatment. This paper examines the evolution of MRSA, analyses the spectrum of diseases produced by this bacterium and presents current prevention and treatment strategies for orthopaedic infections from MRSA

Aim. Skeletal tuberculosis (TB) accounts for up to one third of cases of extra-pulmonary TB but comprises a minority of osteoarticular infection in areas with low TB incidence. Consequently, unexpected cases may receive surgical management targeted at non-tuberculous orthopaedic infections. This study reviewed treatment and outcomes of non-spinal osteoarticular TB to assess outcomes from modern surgical techniques. Method. All patients with a diagnosis of non-spinal osteoarticular TB between 2009–2017 from one tertiary referral centre were included. Retrospective review of surgical intervention, antibiotic treatment and outcome was conducted. Results. Fourteen patients with an average age of 48 (range 20–77) were identified; all were HIV-negative. Articular infections affected 7 patients, including one prosthetic joint infection. Osteomyelitis affecting the carpus, femur, tibia, olecranon and metatarsals was diagnosed in the remaining patients. Only 4 patients had radiological findings consistent with prior pulmonary TB, and only 3 had a history of prior TB or TB exposure. In 2 cases, symptom exacerbation was associated with local steroid injection. Diagnostic biopsy was employed in 5 cases, of whom 4 proceeded to medical management. Diagnosis was made following positive culture in 86% of cases; all TB isolates were fully sensitive. 71% of cases underwent surgical treatment according to best practice for biofilm-forming infection, including excision of osteomyelitis with local antibiotic therapy for three patients, and first-stage excision with spacer implantation for four patients. Quadruple therapy for an average of 8.5 months, range 6–12 months, was administered. Patients were followed up for a mean of 15.2 months. Half of the patients treated with surgery reported ongoing pain at 3 months and 4 patients underwent further surgery for persistent signs of infection (2 for probable persistent TB, 2 for bacterial super-infection). Conclusions. The role of surgical debridement in management of osteoarticular TB is unclear. In patients with a previous history of TB exposure a pre-operative diagnosis of TB could prevent unnecessary surgery and therefore prevent associated post-operative complications including bacterial super-infection and pain. Pre-op biopsy should therefore be considered in all patients with a history of TB exposure

Thermostability is a key property in determining the suitability of local delivery of antibiotics in the treatment of orthopaedic infections. Herein, we aimed to assess the thermal stability and antibacterial activity of ciprofloxacin, ceftriaxone, gentamycine and vancomycine in high temperature conditions. Using a standardized E-test method, minimally inhibited concentration of each antibiotic substance against Staphylococcus aureus cultures were determined. The solutions of antimicrobial drugs ciprofloxacin 2 mg/ml, ceftriaxone 200 mg/ml, gentamycine 40 mg/ml and vancomycine 200 mg/ml were diluted twofold in deionised water. Acquired solutions were divided into three aliquots. The first aliquot was held at 40°C for 30 min in a waterbath, the second and the third aliquots were exposed to 80 and 100°C for 30 min in hot-air sterilizer, respectively. The treated solutions were tested for residual activity against S. aureus using a standardized disk diffusion method. Mediums with untreated antibiotic solutions and S. aureus were used as control. Plates were incubated at 37°C, at which time zones of inhibition (ZoI) were measured to the nearest whole millimeter for 14 days. The investigation indicated that the temperature elevation impacted considerably on antimicrobial activity and antibiotic stability overall. The in vitro temperature-response curves showed that ZoI diameter decreases logarithmically with elevated temperatures. Gentamicin was the only drug that was found to be affected to some extent. Results from the study provides a valuable dataset for orthopaedic surgeons considering local application of antibiotics and methods of antibiotic impregnation

Aim. Most orthopedic infections are due to the microbial colonization of abiotic surfaces, which evolves into biofilm formation. Within biofilms, persisters constitute a microbial subpopulation of cells characterized by a lower metabolic-activity, being phenotipically tolerant to high concentrations of antibiotics. Due to their extreme tolerance, persisters may cause relapses upon treatment discontinuation, leading to infection recalcitrance hindering the bony tissue regeneration. Using isothermal microcalorimetry (IMC), we aimed to evaluate in vitro the presence of persisters in a methicillin-resistant Staphylococcus aureus (MRSA) biofilm after treatment with high concentrations of vancomycin (VAN) and their ability to revert to a normal-growing phenotype during incubation in fresh medium without antibiotic. Moreover, the ability of daptomycin to eradicate the infection by killing persisters was also investigated. Method. A 24h-old MRSA ATCC 43300 biofilm was exposed to 1024 µg/ml VAN for 24h. Metabolism-related heat of biofilm-embedded cells, either during or after VAN-treatment, was monitored in real-time by IMC for 24 or 48h, respectively. To evaluate the presence of VAN-derived “persisters” after antibiotic treatment, beads were sonicated and detached free-floating bacteria were further challenged with 100xMIC VAN (100 µg/ml) in PBS+1% Cation Adjusted Mueller Hinton Broth (CAMHB).. Suspensions were plated for colony counting. The resumption of persister cells' normal growth was analysed by IMC on dislodged trated cells for 15h in CAMHB. Activity of 16 µg/ml daptomycin was assessed against persister cells by colony counting. Results. When incubated with 1024 µg/ml VAN, MRSA biofilm produced undetectable heat, suggesting a strong reduction of cell viability and/or cellular metabolism. However, the same samples re-inoculated in fresh medium produced a detectable and delayed metabolism-related heat signal, similarly to that generated by persister cells. The following exposure to 100xMIC VAN resulted in neither complete killing nor bacterial growth, strongly supporting the hypothesis of a persistent phenotype. IMC analysis indicated that VAN-treated biofilm cells resumed normal growth with a ∼3h-delay, as compared to the untreated growth control. Daptomycin treatment yielded a complete eradication of persister cells selected after VAN treatment. Conclusions. Hostile environmental conditions (e.g. high antibiotic bactericidal concentrations) select for persister cells in MRSA biofilm after 24h-treatment in vitro. A staggered treatment vancomycin/daptomycin allows complete biofilm eradication. These results support the use in clinical practice of a therapeutic regimen based on the combined use of antibiotics to kill persisters and eradicate MRSA biofilms. IMC represents a suitable technique to detect persisters and characterize in real-time their reversion to a metabolically-active phenotype

Orthopaedic infection with bacteria leads to high societal cost and is detrimental to the life quality. Particularly, deep bone infection leading to osteomyelitis results in an inflammatory response whereby localized bone destruction occurs. Current treatments like antibiotic-containing polymethymethacrylate (PMMA) still has the high risk of bacterial resistance. Taking advantages of silver which has antibacterial and anti-inflammatory effect and bioactive collagen, we fabricated a silver nanoparticle (AgNP)-coated collagen membrane by sonication and sputtering. SEM showed good deposition of AgNPs on collagen membrane by both coating methods. The optimal coating concentration was finalized by assessing optimal antibacterial effect against cytotoxicity and finally collagen membrane coated with 1mg/mL AgNPs solution was selected. We also found that the coated collagen membrane demonstrating short-term cytotoxicity within 24 hours with damage to the cell membrane, which was evidenced by MTS and LDH release test, but had no significant influence (p > 0.05) thereafter. The amount of released AgNPs from coated collagen membrane had negligible cytotoxicity (p > 0.05). Confocal laser scanning microscope displayed similar cell morphology in both coated and uncoated collagen membrane. ELISA and qPCR presented the decreased secretion and expression (p < 0.001) of IL-6 and TNF-alpha. Upregulated expression (p < 0.001) of osteogenesis markers (RUNX2, ALP and OPN) could be found and this might be attributed to the modified collagen fibre surface coated by AgNPs. Collectively, the osteogenesis induced by AgNPs demonstrates a promising application in orthopaedic surgery for its use both as an antimicrobial agent, and to enhance bone regeneration

Aim. Here we describe a cohort study to determine the performance of a commercially available Fluorescence In Situ Hybridization (FISH)-kit on samples of 65 consecutive patients suspected of orthopedic implant associated infections (IAI). Culture is routinely used and has a high specificity and sensitivity but requires days to more than a week for slow growing bacteria. FISH results are available within 45–60 minutes and thus specific treatment can start immediately. In addition, previous antibiotic therapy may hinder culture while bacteria may still be detected by FISH. Method. The hemoFISH-kit from Miacom diagnostics (Dusseldorf, Germany) was used on a total of 82 joint aspirates, sonication fluids and tissue samples of 65 consecutive patients to detect and identify possible microorganisms. This FISH-kit contains a universal 16S rRNA probe and species-specific probes for bacteria commonly encountered in blood infections. FISH and culture were compared to the clinical definition of IAI. These definitions were based on the criteria described by Pro-Implant Foundation criteria for IAI after fracture fixation or prosthetic joint infection. If no criteria were described in the literature for a specific IAI then MSIS criteria were used. Results. FISH and culture was done in 33 plain tissue samples, 43 sonication fluid samples and 6 joint aspirates of 65 patients. Results are shown in table 1. In clinical infections FISH provided earlier results in 7 and 2 extra for culture-negative. In 5 IAI-negative cases FISH was false-positive. Conclusions. Faster diagnosis by FISH is appealing, however with a PPV of 64% the hemoFISH-kit is not accurate enough for clinical use. Also, blood and orthopedic infections have different common pathogens, therefor FISH could not identify all of the bacterial strains due to a lack of specific probes. An orthopedic FISH-kit could solve this problem

Aim. Long term use of antibiotics following surgical debridement are the cornerstone of PJI treatment. Due to increasing resistance of bacteria for many first line antibiotics new options are needed. One such option is linezolid known for its low percentage of resistance against many Gram positive bacteria causing PJI. Success rates up to 86% have been reported. At the same time many adverse events (AE) have been described including anemia, thrombocytopenia, gastrointestinal effects and sometimes neuropathy, e.g. irreversible vision loss [1, 2]. Therefore, linezolid use is advised to be limited to a maximum of 28 days. Literature about the effects of prolonged use is currently lacking and therefore this study will aim to determine the safety of long-term (>28 days) linezolid use in patients with orthopedic infections. Methods. We performed a retrospective descriptive study on patient records of orthopedic patients who were treated with linezolid between January 2014 and January 2019 for >28 days. Data were collected from medical charts including co-morbidities, pre-existing liver/kidney dysfunctions, diagnosis, treatment, type of prosthesis, pathogens, adverse events associated with linezolid use and follow up laboratory data. Results. 91 patients treated with linezolid were identified. 46 patients (25 male), mean age 64 (SD 11.49) received long-term linezolid with an average treatment duration of 45.3 (range 29 – 91) days. AE were observed in 32 with gastrointestinal AE's (16 patients) being the most frequent following anemia (7 patients) thrombocytopenia (6 patients), leucopenia (2 patients). One patient reported optic neuritis but no association with linezolid could be confirmed. Linezolid treatment was ended in 7 patients (15.2%) due to AE (predominantly anemia) compared to 14 patients (31.1%) who received short-term treatment (predominantly gastrointestinal AE). Decreased post-surgery hemoglobin levels tended to increase during the first two weeks of linezolid use after which hemoglobin levels showed an average decrease of 0.39 mmol/L between week 2 and week 7 of treatment. Leucocyte and thrombocyte levels showed an average decrease between baseline measures and week 7 of treatment of 2.21∗10. 9. /L and 89.0∗10. 9. /L respectively. AE were resolved after a mean 12 days (range 2–30 days). Conclusions. Long-term linezolid use was not associated with an increase in serious irreversible AE and can therefore be considered generally safe, provided that patients are well monitored considering high drop out in the first weeks mainly due to gastrointestinal AE and anemia during prolonged treatment. These observations help to fill the gap in knowledge about prolonged linezolid use

Aims: To determine the rate of orthopaedic wound infection using ASEPSIS and compare this to the rate of infection as defined by the US Centres for Disease Control (CDC) and the UK Surgical Site Infection Surveillance Service (SSISS). Background: It is a common misconception that reported rates of orthopaedic wound infection are accurate, reliable and reproducible. Most definitions of infection, including CDC and SSISS, are subjective and depend on the interpretation of the surgeon. ASEPSIS. 1. is a method of wound scoring which grades wounds as uninfected, disturbed healing, minor infection, moderate infection and severe infection. ASEPSIS scoring has been proven to be both objective and repeatable. 2. . Method: Over 4 years, 1113 orthopaedic wounds were prospectively evaluated using the CDC definition for surgical site infections, the SSISS definition and the ASEPSIS scoring method. Patients were seen pre-operatively and at 3 and 5 days post-operatively. They also completed a wound surveillance questionnaire at 2 months post-discharge. Results: The overall infection rates were 8% as defined by CDC, 4% as defined by SSISS and 3% as defined by ASEPSIS. Further classification of the wounds as defined by ASEPSIS revealed that 91% of wounds showed no evidence of infection (score < 10), 6.6% showed a disturbance of healing (score 11–20), 2.3% had a minor infection (score 21–30), 0.4% had a moderate infection (score 31–40) and 0.3% had severe infection (score > 40). Conclusion: This study illustrates that accurate wound surveillance is not simple. Different wound infection definitions give very different rates of infection and make comparisons between surgeons and hospitals impossible. We propose that ASEPSIS provides the most accurate and reproducible results and also provides more information with the grading of wound infection. The overall rate of orthopaedic wound infection using the ASEPSIS method is 3%. If all hospitals used this scoring method, more accurate comparisons of infection rates could be made

Aim. Virulent bacteriophages are known to be an effective therapy against various human bacterial infections. The aims of the study are to evaluate i) the killing activity of an antistaphylococcal phage lysate (ASPL), available in the Czech Republic for topical application, against Staphylococcal aureus (Sa) strains isolated in orthopedic infections; ii) the antimicrobial activity of ASPL against biofilm-embedded cells of a methicillin-resistant Sa (MRSA) standard strain. Method. The susceptibility of 25 MRSA and 18 methicillin-sensitive Sa (MSSA) strains to the ASPL was evaluated by spot assay. In addition, susceptibility of four laboratory MRSA strains, including ATCC 43300, ATCC 33591, Mu3 (MRSA/hetero vancomycin intermediate resistant Sa) and Mu50 (MRSA/vancomycin-resistant Sa) was also tested. The activity of ASPL against planktonic and biofilm-embedded MRSA ATCC 43300 was evaluated in real-time by isothermal microcalorimetry. The minimum heat inhibitory concentrations (MHIC) was defined as the lowest antimicrobial concentration leading to the lack of heat flow production after 24h for both planktonic and biofilm-embedded cells. The viability of bacterial cells was assessed by plating and colony counting. The minimum bactericidal concentration (MBC) was defined as the lowest antimicrobial concentration leading the reduction of 3 log CFU compared to the untreated control. Results. Around 34 out of 43 (79%) Sa strains were susceptible to the ASPL, including 17 MRSA and 17 MSSA strains. Both Mu3 and Mu50 (vancomycin intermediate and resistant MRSA, respectively) strains were also susceptible. Microcalorimetric evaluation of the activity of ASPL against planktonic cells of MRSA ATCC 43300 revealed the MHIC and the MBC were 10. 4. PFU/ml and 10. 5. PFU/ml, respectively. ASPL tested at 10. 5. PFU/ml was able to suppress the heat produced by biofilm bacterial cells, although this titer was not able to completely eradicate MRSA biofilm. Conclusions. ASPL showed a broad host spectrum among MRSA and MSSA strains associated with infections on implants, including strains that are resistant to vancomycin as well. ASPL exhibits a lytic activity against planktonic and biofilm MRSA and a titer of phage higher than 10. 5. PFU/ml is needed in order to achieve a complete eradication of MRSA biofilm. In conclusion, the antistaphylococcal phage lysate shows an excellent potential treatment of implant-related infections caused by Sa strains

Aim. S. aureus and S. epidermidis remain the leading biofilm-forming agents causing orthopedic implant-associated infections (OIAI), but other coagulase-negative Staphylococcus (CoNS) with clinical importance is emerging. Besides, few studies have assessed specific genomic traits associated with patient outcome. This is a preliminary descriptive study of phenotypic and genomic features identified in clinical isolates of S. aureus and CoNS isolates recovered from OIAIs patients that progressed to treatment failure. Methods. Ten isolates were identified by matrix-time-of-flight laser-assisted desorption mass spectrometry (MALDI-TOF-MS) and tested for antibiotic susceptibility and biofilm formation. Genotypic characteristics, including, MLST (Multi Locus Sequence Typing), SCCmec typing, virulence and resistance genes were assessed by whole-genome sequencing (WGS) that was performed on an Illumina HiSeq 2500 platform. Bioinformatics analyzes were performed using CGE, PATRIC, VFDB, CARD RGI, SnapGene, BLAST, and PubMLST. S. aureus (215, 260 and 371) isolates belonged to CC5 (ST5 and ST105, spa type t002) and carried SCCmec type I (1B), II (2A) and V(5C2), respectively. Results. They carried multiple resistance genes, with all resistant to methicillin (MRSA), and harboured mecA, blaZ. S. aureus 215 and 371 carried ermA gene and multiple genes for aminoglycosides resistance including aph(3’)-III, ant(9)-Ia, and ant(4)-Ib, and for quinolones. S. aureus 260 also carried resistance genes for tetracycline, quinolones and trimethoprim (dfrC). All MRSA were strong biofilm producers harboring the complete icaADBC and icaR operon, and also carried multiple adhesion and toxin-related virulence genes. Seven CoNS isolates comprising five species (S. epidermidis, S. haemolyticus, S. sciuri, S. capitis and S. lugdunensis) were analyzed, with mecA gene detection in five isolates. S. haemolitycus (95) and S. lugdunensis were unable to form biofilm and did not harbor the complete icaADBCR operon. S. epidermidis (216, 403) and S. haemolyticus (53,95) isolates belonged to the ST2/CC2, ST183, ST9 and ST3, respectively. High variability of adhesion genes was detected, with atl, ebp, icaADBC operon and IS256 being the most common. Conclusions. In conclusion, this study provides insights into the phenotypic and genomic analysis of Staphylococci allowing elucidation of MRSA and CoNS specific features that are associated with treatment failure in OIAIs, including genes associated with biofilm production, and resistance to β-lactam and aminoglycosides

Aim. Fracture related infection (FRI) remains a challenging diagnosis in orthopedic and trauma surgery. In addition to clinical signs and imaging, serum inflammatory markers are often used to estimate the probability of FRI. To what extent serum inflammatory markers can be used to rule out and diagnose FRI remains unclear. The aim of this systematic review was to assess the diagnostic value of the serum inflammatory markers C-reactive protein (CRP), leukocyte count (LC) and erythrocyte sedimentation rate (ESR) in suspected fracture related infection. Method. PubMed, Embase and Cochrane databases were searched for all articles focusing on the diagnostic value of CRP, LC and ESR in FRI. Studies on other inflammatory markers or other types of orthopedic infection, such as periprosthetic and diabetic foot infections, were excluded. For each serum inflammatory marker, all reported sensitivity and specificity combinations were extracted and graphically visualized. Average estimates were obtained using bivariate mixed effects models. This study utilized the QUADAS-2 criteria and was reported following the PRISMA statement. Results. The search resulted in 8280 articles, of which seven were eligible for inclusion. One study was excluded after quality assessment. CRP was reported in all included studies, with sensitivity ranging from 60.0 to 100.0% and specificity from 34.3 to 85.7%. Five of these studies were pooled. The average pooled sensitivity and specificity of CRP were, respectively, 77.0% (95% CI 66.5–85.0%) and 67.9% (95% CI 38.7–87.6%). LC was reported in five studies. Sensitivity ranged from 22.9 to 72.6% and specificity from 73.5 to 85.7%. The results of four of these studies were pooled, resulting in a 51.7% (95% CI 27.2–75.5%) sensitivity and 67.1% (95% CI 19.3–50.2%) specificity. ESR was reported in five studies. Sensitivity and specificity ranged from 37.1 to 100.0% and 59.0 to 85.0% respectively. Three of these studies were pooled, showing a 45.1% (95% CI 37.8–52.6%) sensitivity and 79.3% (95% CI 71.7–85.2%) specificity of ESR. Four studies analyzed the combined value of inflammatory markers, reporting an increased diagnostic accuracy. These results could not be pooled due to heterogeneity. Conclusions. The serum inflammatory markers CRP, LC and ESR are insufficiently accurate to diagnose FRI. These markers cannot rule out the presence of FRI, but they may be used as a suggestive sign in the diagnosis of FRI

Aim. The incidence of orthopaedic methicillin-resistant staphylococcus aureus infections is increasing. Vancomycin may therefore play an increasingly important role in orthopaedic perioperative antimicrobial prophylaxis. Adequate antimicrobial concentrations at target site is essential for prevention of orthopaedic infections. Current studies investigating perioperative bone and soft tissue concentrations of vancomycin are sparse and challenged by a lack of appropriate methods. The aim of this study was therefore to assess the concentration of vancomycin in plasma, subcutaneous tissue and bone after single dose administration using microdialysis (MD) in patients undergoing total knee replacement. Method. 1,000 mg of vancomycin was postoperatively administered intravenously over 100 minutes to 10 male patients undergoing primary total knee replacement. Vancomycin concentrations in plasma, subcutaneous tissue (SCT), cancellous and cortical bone were measured the following 8 hours. MD was applied for sampling in solid tissues. The vancomycin concentration in MD-samples was determined using ultra-high performance liquid chromatography, whilst the free plasma concentration was determined using a chemistry analyzer*. Results. For all extravascular tissue, an impaired penetration was demonstrated, with lower area under the concentration-time curve (AUC) compared to free plasma. The lowest AUC was found in cortical bone. For all tissues, tissue penetration expressed as the ratio of the area under the concentration–time curve from 0 to the last measured value (AUC0-last tissue/AUC0-last plasma) were below 0.5. The time to a mean clinically relevant minimal inhibitory concentration (MIC) of 2 mg/L were 3, 36, 27 and 110 min for plasma, SCT, cancellous and cortical bone, respectively. As opposed to the other compartments, a mean MIC of 4 mg/L was not reached in cortical bone. The AUC0-last and peak drug concentrations (Cmax) for SCT, cancellous and cortical bone were lower than those of free plasma. The time to Cmax was higher for all tissues compared with free plasma. Conclusions. Penetration of vancomycin to bone and SCT was found to be impaired and delayed in male patients undergoing total knee replacement surgery. Adequate perioperative vancomycin concentrations may not be reached at target site using standard prophylactic dosage. *Cobas c501

Objectives. Thermal stability is a key property in determining the suitability of an antibiotic agent for local application in the treatment of orthopaedic infections. Despite the fact that long-term therapy is a stated goal of novel local delivery carriers, data describing thermal stability over a long period are scarce, and studies that avoid interference from specific carrier materials are absent from the orthopaedic literature. Methods. In this study, a total of 38 frequently used antibiotic agents were maintained at 37°C in saline solution, and degradation and antibacterial activity assessed over six weeks. The impact of an initial supplementary heat exposure mimicking exothermically curing bone cement was also tested as this material is commonly used as a local delivery vehicle. Antibiotic degradation was assessed by liquid chromatography coupled to mass spectrometry, or by immunoassays, as appropriate. Antibacterial activity over time was determined by the Kirby-Bauer disk diffusion assay. Results. The heat exposure mimicking curing bone cement had minimal effect on stability for most antibiotics, except for gentamicin which experienced approximately 25% degradation as measured by immunoassay. Beta-lactam antibiotics were found to degrade quite rapidly at 37°C regardless of whether there was an initial heat exposure. Excellent long-term stability was observed for aminoglycosides, glycopeptides, tetracyclines and quinolones under both conditions. Conclusions. This study provides a valuable dataset for orthopaedic surgeons considering local application of antibiotics, and for material scientists looking to develop next-generation controlled or extended-release antibiotic carriers. Cite this article: E. Samara, T. F. Moriarty, L. A. Decosterd, R. G. Richards, E. Gautier, P. Wahl. Antibiotic stability over six weeks in aqueous solution at body temperature with and without heat treatment that mimics the curing of bone cement. Bone Joint J 2017;6:296–306. DOI: 10.1302/2046-3758.65.BJR-2017-0276.R1

The aim of our project is to develop resorbable nanostructured composite layer with controlled elution of antibiotics for implants survival rate enhancement. The nanostructured layers are expected to be used especially in the case of known systemic or local (joint) inflammation. This layer can provide a bone tissue/implant (titanium alloy) bioactive interface improving the physiological healing process and eliminating the risk of bacterial orthopedic infections. The main aim of this study was to verify whether the local concentration of released vancomycin exceeded the minimum inhibitory concentration (MIC) for vancomycin-resistant Staphylococcus aureus (VRSA, >16 mg/l). The layer is composed of collagen (type I, isolated form calf skin), hydroxyapatite nanoparticles and vancomycin hydrochloride (10 wt%). The stability of collagen was enhanced by EDC/NHS cross-linking. The in vitro release of vancomycin and crystalline degradation products from optimally cross-linked layers was investigated. An elution method and a high performance liquid chromatographic assay were employed to characterize the in vitro release rates of the vancomycin and its crystalline degradation antibacterial inactive products over a 21-day period. During the whole experimental period, the level of released vancomycin was high above the MIC for VRSA. The maximum average concentration was obtained between day 4 and day 8 and it reached 265 mg/l. At the end of the experiment (day 21), an average concentration of 104 mg/l was detected. Our study confirmed the prophylactic effects of studied vancomycin-loaded nanostructured layers

Aim. Photodynamic therapy (PDT) requires a photosensitiser, a light source of an appropriate wavelength, and the presence of molecular oxygen. Once stimulated to its excited phase by the light, the photosensitiser reacts with oxygen to form free radicals of ‘singlet oxygen’ which is cytotoxic to microorganisms. We aim to demonstrate the effectiveness of PDT as an in-vitro antimicrobial technique against Staphylococcus aureus, Methicillin resistant Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Acinetobacter bauminii. This will form the scientific basis for further animal and human studies assessing PDT for treatment of periprosthetic infections, septic arthritis, and open fractures. Method. A PDT treatment protocol was devised using lawns of bacteria on agar plates. PDT was targeted towards the bacteria and the remaining microorganisms were quantified using a serial dilution technique. In order to assess the ability of photodynamic therapy to target biofilms on metallic implants, biofilms were cultured on polished titanium and hydroxyapatite-coated titanium discs and subjected to PDT. Results. Reductions in bacterial colony forming units of up to 7 log were achieved using PDT. The figure is a box plot representing a comparison of the amount of biofilm Pseudomonas aeruginosa (cfu/ml) remaining on the polished titanium disc and hydroxyapatite-coated titanium disc following treatment with photodynamic therapy. (MB+/-: photosensitizer present/absent; L+/-: laser present/absent). Conclusions. PDT has long been used in dermatology and dentistry as an antimicrobial technique. Its potential for treating orthopaedic infections has not yet been investigated. This study demonstrates potential for PDT as an antimicrobial technique in the treatment of bacteria commonly found in periprosthetic infections, septic arthritis, and open fractures. This in-vitro work lays the foundations for future animal and clinical studies. We envision PDT being used as an adjunct to antibiotics in treatment of these conditions, helping prevent ongoing infection, and the development of resistance

Of the 6075 patients enrolled in EU-CORE registry, 206 patients had orthopaedic device-related infections. Significant underlying diseases were reported in 71% patients, most frequently cardiovascular disease (38%). The common sites of infection were knee (40%) and hip (33%). Among the 170 patients with available culture results, 135 (79%) were positive. Coagulase-negative staphylococci (CoNS, 44%) and Staphylococcus aureus (43%, of those 47% were methicillin resistant) were the most commonly isolated pathogens. Daptomycin was used empirically in 48% patients and as second-line therapy in 67% patients. During daptomycin therapy, 67% patients had undergone surgery (debridement, 61%; removal of foreign device, 39%; incision and drainage, 9%). Over half of the inpatients (54%) received concomitant antibiotics. Daptomycin was most frequently prescribed at a dose of 6 mg/kg/day (48%), with a median duration of therapy of 16 (range, 1–176) days. The overall clinical success rate was 85%, and was similar whether daptomycin was administered as first- or second-line therapy. The success rates achieved for infections caused by S. aureus and CoNS were 86% and 83%, respectively. Among the 79 patients who entered the long-term follow-up, 85% had a sustained response. Adverse events (AEs) and serious AEs possibly related to daptomycin were reported in 4.4% and 1.9% patients, respectively. Results from this real-world clinical experience showed that daptomycin is an effective and well-tolerated treatment option for orthopaedic device-related infections with a high success rate up to 2 years of follow-up

Bone and joint infections are not only common but their management can be technically complex. They carry significant healthcare costs and are a daunting experience for patients [1]. Frequently, multiple operations are required in order to treat the infection. Each surgical intervention usually results in greater bone loss, worsening skin and soft tissue scarring and increasingly diverse and resistant micro- organisms [2]. Specialist bone infection units involving highly integrated orthopaedic and plastic surgery, as well as infection physicians, may improve patient outcomes [3–4]. However, it is difficult to determine the hierarchy of factors contributing to outcome of treatment. This problem is confounded by a lack of structured, prospective data collection in many units around the world. In 2014, we designed a modular database which allows collection of patients’ details, components of the disease, the treatment, microbiology, histology, clinical outcome and patient-reported outcome measures (PROMS). The registry was implemented in November 2014 and has already demonstrated its function as a Hospital-wide service evaluation tool. Over 200 patients have been referred to the unit and their baseline demographic information registered. Their progress through the bone infection unit patient pathway is prospectively monitored with use of the registry and data collection ongoing. We aim to present the preliminary clinical outcomes of these 200 patients including surgical procedures performed, key microbiology results, antibiotic treatment regimens and patient reported outcomes. Our goal is to demonstrate that a bone infection registry is an integral part of infection management clinical practice. It can be used for designing service provision, assist in allocating healthcare resources and expand the evidence base for specialist bone infection units in managing complex orthopaedic infections

Introduction: In recent years the implementation of sonication in the diagnosis of orthopaedic implant infections has improved the detection of subclinical infections. With the use of sonication of removed orthopaedic material we can detect the presence of biofilm. The method has already shown encouraging results, especially in cases of preoperative antibiotic therapy. Aim: The aim of the study was to detect infections of orthopaedic material using both sonication and standard diagnostic methods, and to compare the obtained results of both methods. For the purpose of the study we sonicated all explanted material at revision surgery and cultured the obtained samples. During revision surgery soft tissue biopsies were collected and analyzed using standard microbiologic methods. The results were compared, analyzed and additional therapy was applied, if an infection of the material was proven. During the period from September 2009 to the end of March 2014 we studied 249 cases (198 patients) of revision surgery (166 cases of revision hip arthroplasty, 53 cases of revision knee arthroplasty, 13 cases of revision foot surgery, 17 cases of revision spine surgery). Of studied cases infection was proven in 20 (8,0%) cases by soft tissue biopsies only, 90 cases (36,1%) were diagnosed both by soft tissue biopsies and sonication, 45 cases (18,1%) were diagnosed only by sonication of explanted prosthetic material and in 94 cases (37,8%) all results were negative. The statistical analysis has shown statistically significant (p<0,05) improvement of infection detection using sonication. According to our experience the implementation of sonication has shown an improvement in the diagnosis of orthopaedic implant infections. Despite certain limitations, sonication should be considered in doubtful cases of revision surgery. The use of sonication should be emphasized in cases of preoperative antibiotic treatment

Melioidosis is an uncommon infection caused by a Gram-negative bacillus, Pseudomonas pseudomallei. Only a few case reports of orthopaedic infection have been published in English, and most were of isolated septic arthritis or secondary to melioidosis of another organ. We have reviewed ten patients with localised melioidotic osteomyelitis; six had underlying conditions. We discuss the importance of obtaining a bacteriological diagnosis, and of surgical debridement as well as appropriate antibiotic therapy

Direct observations have shown that the bacteria and fungi that cause device-related and other chronic infections grow in well developed biofilms on the surfaces of biomaterials and of compromised tissues. This mode-of-growth confers on the microorganisms an inherent resistance to host defenses, and antibiotic therapy, and makes these infections very difficult to detect because biofilm bacteria do not produce colonies when plated on the agar media used in routine cultures. We have initiated two comprehensive studies of total joint prostheses, and of non-unions secondary to trauma, in which we use DNA-based (Ibis and 454) methods for the detection of bacteria and an iterative process in which we locate and visualize biofilms using FISH probes and confocal microscopy. The DNA-based detection system confirms culture results, but detects more organisms and determines their sensitivity to antibiotics, and appears to be useful in the management of both types of infection. The use of confocal microscopy and FISH probes to visualize and map biofilms, in relation to orthopedic hardware and affected tissues, confirms the Ibis data and provides useful insights into the etiology of orthopedic infections

The role of biofilm in pathogenesis of several chronic human infections is widely accepted, as this structure leads pathogens to persist among the human body, being protected from the action of antibacterial molecules and drugs (1). It has been estimated that up to 65% of bacterial infections are caused by microorganisms growing in biofilms (2). Moreover, biofilm is involved in device-related orthopaedic bacterial infections, which are unaffected by vaccines and antibiotic therapies, constituting a serious problem for the human health care. The aim of the present work was to evaluate the anti-biofilm action of a selected and patented lactobacillus strain (MD1) supernatant, both on the in-formation- biofilm and on mature biofilm produced by pathogenic bacteria. MD1 was grown in BHI for 48 h at 37°C. After incubation, the sample was centrifuged for 5’ for 14,000 × g and the supernatant previously filtered and treated in order to obtain the anti-biofilm compounds (Special Supernatant – SS) was collected. Staphylococcus aureus and Pseudomonas aeruginosa strains were grown in BHI for 24h at 37°C. The anti-biofilm ability of the tested SS – lactobacillus strain was evaluated by a spectrophotometric method according to Christensen at al., following the incubation of pathogens and the “mature biofilm” with the lactobacillus supernatant. Confocal Laser Scanning Microscopy was used to confirm the data obtained from Crystal Violet Assay. After the incubation of the SS with pathogens and mature biofilm, the formation of biofilm was inhibited and a significant disruption of the mature biofilm was observed. Interestingly, the same properties were observed also when the SS pH was neutralized to pH 6.5. In particular, the reduction of biofilm production and the disruption of mature biofilm was about 50–70% for all microorganisms. The SS lactobacillus strain MD1 exhibited a relevant antibiofilm action against mature and in-formation-biofilm produced by S. aureus and P. aeruginosa strains tested in the study. Moreover, the antibiofilm action has been observed to be pH-independent, as when the supernatant was neutralized to pH 6.5, the reduction of pathogenic biofilm has been still observed. These promising results highlighted the possibility to use this SS-lactobacillus anti-biofilm property to develop a cost-effective and safety treatment able to reduce the impact of pathogenic biofilm on device-related orthopaedic bacterial infections

We used 99 strains of organisms representative of orthopaedic infections to examine the effectiveness of a bone cement containing tobramycin, employing a modified in vitro Kirby-Bauer susceptibility model. The spectrum was broad, including Gram-positive and Gram-negative aerobic organisms, anaerobes and mycobacteria. Simplex P with added tobramycin was effective against most of the strains, including those which are resistant to typical systemic levels of tobramycin. Although direct correlation between in vitro and in vivo results is difficult, the study showed that tobramycin is stable to the exothermic polymerisation of the cement, and that it is released from the surface of the cement at concentrations high enough to inhibit the growth of most organisms which may be encountered after joint arthroplasty

Corynebacterium Jeikeium is a pathogen rarely involved in orthopaedic infections. Till date only 14 cases of osteomyelitis are described in the literature, envolving the tibia, foot and prosthethic (hip and joint) infection. To our knowledge, Corynebacterium Jeikeium as not been reported as an infectious agent of the spine. Our goal is to describe a case of scoliosis surgical site infection by a Corynebacterium Jeikeium specimen. This is a retrospective and descriptive case report based on data from clinical records, patient observation and analysis of complementary exams. We present a 24 year old female with a history of premature birth, West syndrome, spastic cerebral palsy and spina bifida. She was sent to our consult for evaluation of dorsolombar scoliosis. In October of 2014, she was submitted to surgery – posterior spine arthrodesis and instrumentation (D10 to L5) with bilateral pedicle screws and two chromium-cobalt bars. The early post-operative period was without complications. She was discharge at the seventh day of internment and was seen, fifteen days postoperative, at the consultation office, where the dressings were changed, with no signs of surgical site infection. One month post-operative, she recurs to the office because of an apparent seroma at the surgical site wound. There was no reference to fever or other signs of local/systemic infection. A swabbing of the wound was done and the patient was medicated with Ciprofloxacine, 500mg 12/12 hours – the culture came back negative. Seven days later she was seen again, maintaining the seroma with purge of a serous-aspect fluid. Antibiotic therapy was maintained and another swabbing was collected – culture came back negative. Because of suspected surgical site infection, she was re-operated at December of 2014. Surgical wound debridement was performed; three tissue samples and one exudate were sent to the microbiological department. In all samples but one was identified a Corynebacterium Jeikeium. No sensitivity test was performed. Intravenous Vancomicine, 1 gram 12/12 hours was started and maintained during 8 days. Eleven days post-operative she was discharged with oral Vibramicin, 100 milligrams 12/12 hours for two weeks. She is currently being followed at the doctor's office, with no sign of reinfection of the surgical site. This is the first reported case describing an infection of the spine by a Corynebactereium Jeikeium. Isolation of the causative agent of infection and literature-based directed antibiotherapy are important for a successful outcome

In many countries Haemophilus influenzae type b (Hib) is the second most common cause of septic arthritis in children. In Finland large-scale immunisation against Hib using conjugate vaccines began in 1986, four years after a multicentre prospective study of orthopaedic infections in children had started. Since 1982, including six years before and ten after starting routine Hib vaccination, there has been a major change in the pattern of septic arthritis. From 1982 to 1988, 32 of 61 cases (53%) were caused by staphylococci, 22 (36%) by Hib and 7 (11%) by other bacteria. Since 1988, Hib infection has disappeared, and one-third of cases of childhood septic arthritis has been eliminated. This change has allowed us to reduce initial antimicrobial therapy for such children to cover only Gram-positive cocci. The more limited treatment is safer, reduces cost, and simplifies treatment

Introduction. The role of diathermy in orthopaedic surgical practice has increased since its introduction. It is widely used for underlying tissue dissection, cutting, and haemostasis. Previous studies have compared electrosurgical and scalpel incisions in terms of wound infection, wound-related pain, and blood loss. There are well documented hazards associated with diathermy use including burns injury, electrocution, hypoxic stress, inhalation of diathermy plume, and gene mutation. No single study to date has focused on the potential for diathermy tips to cause wound contamination and infection. We sought to identify whether diathermy tips could be possible sources of infection in orthopaedic procedures. Objectives. To determine the prevalence of bacterial contamination of diathermy tips during orthopaedic surgery and to assess any correlation with surgical site infections. Methods. From July 2013 to September 2013, the diathermy tips from 86 consecutive orthopaedic procedures using diathermy were cultured using direct and enriched media. None of the diathermy tips were used for the skin incision. All patients underwent an orthopaedic procedure for a non-infected condition. For each procedure an unused control diathermy tip was placed on the instrument table at the beginning of the procedure and processed similarly. All patients were followed for any postoperative complications. Results. 108 diathermy tips from 86 orthopaedic procedures were cultured. None of the tips cultured directly on blood agar demonstrated bacterial growth. Following enrichment culture, 6 (5.6%) of the procedure diathermy tips and 1 (0.92%) of the control tips demonstrated bacterial growth. Coagulase-negative staphylococci (83.3%) and proprionibacterium (16.7%) were cultured from the tips. 1 of the patients who had bacterial growth from the diathermy tip developed a superficial surgical site infection. Conclusions. Surgical site infections contribute substantially to orthopaedic surgical morbidity and mortality each year. The prevention of these infections encompasses careful operative technique, preoperative antibiotics, and a number of important measures to minimize the risk of bacterial contamination posed by operative staff, the operating theatre environment, and the patient's endogenous skin flora. Identifying potential bacterial sources is an important component of surgery. The two bacteria cultured in our study (coagulase-negative staphylococci and proprionibacterium) are both well known major culprits in orthopaedic infections, responsible for up to 70% of early and late peri-prosthetic infections. Our study suggests diathermy tips and the tissue coagulated by its use may not be as sterile as previously thought. There may be benefit in changing the diathermy tips during orthopaedic procedures as they may represent a possible source of bacterial contamination

The role of diathermy in orthopaedic surgical practice has increased since its introduction. It is widely used for underlying tissue dissection, cutting, and haemostasis. Previous studies have compared electrosurgical and scalpel incisions in terms of wound infection, wound-related pain, and blood loss. There are well documented hazards associated with diathermy use including burns injury, electrocution, hypoxic stress, inhalation of diathermy plume, and gene mutation. No single study to date has focused on the potential for diathermy tips to cause wound contamination and infection. We sought to identify whether diathermy tips could be possible sources of infection in orthopaedic procedures. To determine the prevalence of bacterial contamination of diathermy tips during orthopaedic surgery and to assess any correlation with surgical site infections. From July 2013 to September 2013, the diathermy tips from 86 consecutive orthopaedic procedures using diathermy were cultured using direct and enriched media. None of the diathermy tips were used for the skin incision. All patients underwent an orthopaedic procedure for a non-infected condition. For each procedure an unused control diathermy tip was placed on the instrument table at the beginning of the procedure and processed similarly. All patients were followed for any postoperative complications. 108 diathermy tips from 86 orthopaedic procedures were cultured. None of the tips cultured directly on blood agar demonstrated bacterial growth. Following enrichment culture, 6 (5.6%) of the procedure diathermy tips and 1 (0.92%) of the control tips demonstrated bacterial growth. Coagulase-negative staphylococci (83.3%) and proprionibacterium (16.7%) were cultured from the tips. 1 of the patients who had bacterial growth from the diathermy tip developed a superficial surgical site infection. Surgical site infections contribute substantially to orthopaedic surgical morbidity and mortality each year. The prevention of these infections encompasses careful operative technique, preoperative antibiotics, and a number of important measures to minimize the risk of bacterial contamination posed by operative staff, the operating theatre environment, and the patient's endogenous skin flora. Identifying potential bacterial sources is an important component of surgery. The two bacteria cultured in our study (coagulase-negative staphylococci and proprionibacterium) are both well known major culprits in orthopaedic infections, responsible for up to 70% of early and late peri-prosthetic infections. Our study suggests diathermy tips and the tissue coagulated by its use may not be as sterile as previously thought. There may be benefit in changing the diathermy tips during orthopaedic procedures as they may represent a possible source of bacterial contamination

Abstract. OBJECTIVES. Staphylococcus aureus is one of the most common pathogens in orthopaedic biomaterial-associated infections. The transition of planktonic S. aureus to its biofilm phenotype is critical in the pathogenesis of biomaterial-associated infections and the development of antimicrobial tolerance, which leads to ineffective eradication in clinical practice. This study sought to elucidate the effect of non-lethal dispersion on antimicrobial tolerance in S. aureus biofilms. METHODS. Using a methicillin-sensitive S. aureus reference strain, the effect of non-lethal dispersion on gentamicin tolerance, cellular activity, and the intracellular metabolome of biofilm-associated bacteria were examined. Gentamicin tolerance was estimated using the dissolvable bead biofilm assay. Cellular activity was estimated using the triphenyltetrazolium chloride assay. Metabolome analysis was performed using tandem high-performance liquid chromatography and mass spectrometry. RESULTS. Non-lethal dispersion of biofilm-associated S. aureus was associated with a four-fold reduction in gentamicin tolerance and a 25% increase in cellular respiration of both dispersed and adherent cells. Metabolome analysis found non-lethal dispersion reduced intracellular levels of L-ornithine and L-proline, with increased levels of cyclic nucleotides (p<0.05) in both liberated cells and the remaining biofilm-associated bacteria. These metabolomic changes have previously been shown to be associated with inactivation of the carbon catabolite repression mechanism, which is a key regulatory gatekeeper in the cellular resuscitation of dormant S. aureus cells. CONCLUSION. The metabolomic pipeline described in this study presents a valuable tool in the elucidation of molecular mechanistic pathways in biofilm pathogenesis. Kreb's cycle reactivation, through the carbon catabolite repression regulatory mechanism, has been shown to be associated with the reversal of biofilm-associated gentamicin tolerance. Understanding of the biosynthetic changes associated with the biofilm state will assist in the discovery of novel therapeutic targets in the management of biomaterial-related infections. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

First generation molecular diagnostics based on PCR suggested that the routine culture of bacteria was inadequate for the detection of many pathogens, particularly after antibiotic treatment or when associated with chronic infection and biofilm growth. These techniques, however, suffered from their own problems. False negative results were caused by inhibitors of the PCR process and by the overly specific nature of most simplex assays which require an a priori assumption on the part of the investigator as to which species to test for. False positives resulted from contamination, or carryover, of amplified DNA. Recently several new technologies have been developed and have resulted in “next generation” tests that overcome the problems associated with the earlier methods. We will provide an overview of two of these technologies and present our experience in their application to the diagnosis of orthopedic infections associated with arthroplasties and external fixations. 454-based deep 16S rDNA sequencing provides for a comprehensive and quantitative analysis of all bacterial species present in clinical specimens regardless of whether the species present have been previously identified. The results of this test can be used to improve the specificity of other tests such as the Ibis Universal Biosensor. The Ibis Universal Biosensor T-5000 system uses a highly multiplex PCR front end which is coupled to a highly sensitive electron spray ionization (ESI) time-of-flight (TOF) mass spectrometer (MS) which provides for the exact base composition of the amplified DNA permitting species and even strain-specific identification of bacterial and fungal pathogens through an interface with a massive DNA sequence database. This system therefore provides both great breadth of coverage, with exquisite specificity. Moreover, this system can identify multiple species within a specimen providing a rapid analysis of polymicrobial infections

Aim. To assess the role of Tc-99m labelled anti granulocyte monoclonal antibody Fab' fragment (Sulesomab) in the diagnosis of bone and joint infections. Methods. We analysed the results of 95 patients referred with a clinical suspicion of bone and joint infections. There were 47 male and 48 female patients with a mean age of 60 years (range=16 to 89). Referrals were made for suspected infection of prosthetic total joint replacements (38), long bones (32), primary joints (12) and feet (13). Sulesomab imaging was done with 650 MBq of 99mTcSulesomab. The final diagnosis was determined by conclusive microbiology, culture and/or histology, intra-operative findings, aspiration, complementary investigations like CT/MRI and long term clinical follow-up. The findings of 99mTcSulesomab images were compared with the clinical outcome to arrive at the decision of True Positive/ False positive/ True negative/ False negative results. Using the above definitions sensitivity, specificity and diagnostic accuracy of 99mTcSulesomab for suspected bone and joint infection were calculated. Results. 58/95 patients had normal or equivocal blood results. Plain radiographs revealed no abnormality or were inconclusive of infection in more than half of the patients. Outcome classification revealed 29 true positives, 56 true negatives, 9 false positives and one false negative. The overall sensitivity was 96.66% and specificity 86.15% with a negative predictive value of 98.24%. The individual sensitivity and specificity of each category was compared. Diagnostic accuracy for long bone infections (96.87%) was the highest than any other group. Conclusion. 99mTc Sulesomab imaging has a high sensitivity and specificity in a heterogeneous group of orthopaedic bone and joint infections. With a high negative predictive value, this test seems to be useful in excluding orthopaedic infection rather than confirming it

Introduction: Postoperative infection rates following endoprosthetic surgery are currently around 11%. In comparison with routine orthopaedic infection rates (0.5–2.0%) this is high and warrants investigation. Antibiotic prophylaxis at the time of surgery has been proven to reduce postoperative infection rates. We are looking to see if this is being administered correctly. Methods: We sought the records of 183 patients undergoing primary EPRs at the ROH within an 18 month period. We were unable to locate 57 (69%) leaving 126 to analyse. We recorded details of the surgeon conducting the operation, the perioperative antibiotics administered, and whether the patient developed a SSI postoperatively. We grouped the patients into three groups, those who received the correct regime as defined by the department (1.5g Cefuroxime at induction followed by 3 doses of 750mg Cefuroxime at 8, 16 and 24 hours post-op), those who received cefuroxime but in an incorrect regime and those not receiving cefuroxime. We further looked at the frequency individual surgeons were administering the correct antibiotics. Finally we applied a Chi-squared anaylsis to see if there was a significant difference in infection rates between those receiving the correct antibiotics and those not. Results: 52 patients (41%) received cefuroxime in the correct regime, 27 (22%) received cefuroxime in an incorrect regime and 47 patients (37%) did not receive cefuroxime. The different surgeons at the hospital administered the correct regime in varying frequencies (0%–65%). Giving the correct antibiotic dose is associated with a reduction in the incidence of postoperative SSI (0.1> p> 0.05). Discussion: A large proportion of EPRs are carried out without the recommended prophylactic antibiotic cover. The rate of correct antibiotic administration varies widely between surgeons from the same department. This lack of appropriate prescribing could be contributing to a high rate of postoperative infection and therefore needs addressing

Background:. Early diagnosis of septic arthritis and osteomyelitis in children is essential to prevent long term sequelae. The diagnosis for these orthopaedic emergencies can be difficult and challenging especially in infants. Standard blood tests used for diagnosis have a low specificity. Procalcitonin (PCT) is significantly elevated in bacterial infections and remains low in viral infections and inflammatory conditions. Good positive predictive values for PCT have been obtained in various studies used in paediatric infections, but limited studies have examined the role in orthopaedic infections. Aims:. To introduce PCT testing in the work up of Septic Arthritis (SA) and Acute Osteomyelitis (OM) and to see if the test is useful in the diagnosis. Also to determine whether 0.2 ng/ml is a suitable cutoff level as indicated by previous studies. Method:. All children under 14 years presenting with signs and symptoms of SA/OM from 1 June 2009 to 31 June 2010 were subjected to standard blood tests with addition of PCT and compared to a control group. The definitive diagnosis was made by microbiologic examination obtained in theatre. A PCT cut-off level of 0.2 ng/mL was used. Results:. A total number of 33 patients were included in the study. Eight patients were diagnosed as OM, 4 as SA and 21 had another diagnosis. Staphylococcus aureus was the most common organism isolated in this series with no resistant organisms seen. In the SA/OM Group 11 of the 12 patients had an increased PCT level and 4 in the other diagnosis group had raised PCT. The calculated sensitivity of PCT was 92% with a confidence interval of 62–100% and the specificity was 81% with a confidence interval of 58–95%. In this study the sensitivity of CRP was 100% while the specificity 26%. The positive predictive value for PCT in this study was 73% and the negative predictive value was 94%. The accuracy for PCT in Septic Arthritis and Osteomyelitis in this study was 85%. Conclusions:. The calculated sensitivity and specificity in this study has shown that PCT testing can aid in the diagnosis of SA/OM in children using 0.2 ng/ml as cut-off level. PCT was more specific for bacterial infections in this study than CRP. Further research is needed with larger numbers to conclusively prove that this specific cut-off for PCT is significant

Introduction: Standard therapy for orthopedic device infections includes a two-stage exchange and prolonged antimicrobial therapy. In a subgroup of patients, retention of the device seems to be an effective alternative. Methods: In a prospective study we evaluated treatment efþcacy of orthopedic device infections with implant retention. Inclusion criteria were: early manifestation, stable implant, known pathogen, susceptibility of staphylococci to quinolones and rifampin, good condition of soft tissue. Initially, intravenous antimicrobial therapy was given for 2 weeks, followed by oral treatment for 10 weeks (knee prostheses for 6 months). Results: From January 1999 through June 2002, 19 patients were included: hip prosthesis (9), knee prostheses (6) and internal þxation devices (4). Isolated pathogens were: staphylococci (14), streptococci (4), enterococci (1), and Propionibacterium acnes (1). Open debridement with device retention was performed in 13 patients; the remaining 6 patients were treated with antibiotics only. After initial 2-week intravenous therapy, staphylococcal infections were treated with oral ciproßoxacin 750 mg bid + rifampin 450 mg bid, streptococcal and enterococcal infections with oral amoxicillin 750 mg tid and the P. acnes-infection with oral clindamycin 600 mg tid. 12 of 16 patients were followed for at least 24 months. 10 (83%) had no symptoms or signs of infection at follow-up, 2 (17%) had a relapse Conclusion: In carefully selected patients, device retention with antimicrobial treatment for 3–6 months may be an effective approach

Introduction: Staphylococci are a well recognized cause of orthopaedic implant infections, due to their ability to produce biofilms, that limit antibiotic and host defence capabilities. To detect serum IgM levels against staphylococcal slime polysaccharide antigens (SSPA), an original immunoenzymatic assay has been previously developed and tested in patients affected by staphylococcal vascular graft infections [The Lancet • 359:2166–2168, 2002]. This is the first report on the efficacy of this serum ELISA testing of anti-SSPA IgM in Staphylococcal Orthopaedic Implant-Related Infections (SOIRI). Methods: SSPA is extracted and purified using a suitable original patented bacterial strain and method. The immunoenzymatic assay to detect anti-SSPA IgM was performed on sera collected from 53 patients with joint prosthesis (24 hips, 17 knees, 2 shoulders). Each serum sample was tested in triplicate in two different assays and values are expressed as means. Main exclusion criteria were: time from implant < 6 months, rheumatoid arthritis, concurrent known infections. The study was approved by the local ethical committee and all patients gave their informed consent. 28 sera were collected from controls (patients with uninfected joint prosthesis, as judged from clinical, laboratory and radiological data) and 25 sera were obtained from patients with known SOIRI, sustained by S. aureus or coagulase-negative Staphilococci (CNS) (positive joint aspiration and/or intra-operative cultural examination). Results: 25 patients were classified as true negative, 21 true positive, 4 false negative, 3 false positive. Test efficacy calculation provided the following Results: sensitivity: 84 %, specificity: 89.3 %, positive predictive value: 87.5 %, negative predictive value: 86.2 %. Discussion and Conclusion: Anti-SSPA IgM immunoenzymatic assay showed interesting sensitivity and specificity in this preliminary prospective study. The test appear as an innovative, user-friendly, cheap and non-invasive diagnostic tool for orthopaedic implant-related infections. Further studies should be devoted to better define the cut-off value, testing reproducibility and standardization for large scale application

Introduction and Objective. Found in bone-associated prosthesis, Cutibacterium acnes (C. acnes) is isolated in more than 50% of osteoarticular prosthesis infections, particularly those involving shoulder prostheses. Ongoing controversies exist concerning the origin of C. acnes infection. Few reports construct a reasonable hypothesis about probable contaminant displaced from the superficial skin into the surgical wound. Indeed, despite strict aseptic procedures, transecting the sebaceous glands after incision might result in C. acnes leakage into the surgical wound. More recently, the presence of commensal C. acnes in deep intra-articular tissues was reported. C. acnes was thus detected in the intracellular compartment of macrophages and stromal cells in 62.5% of the tested patients who did not undergo skin penetration. Among bone stromal cells, mesenchymal stem cells (MSCs) are predominantly found in bone marrow and periosteum. MSCs are the source of osteogenic lines of cells capable of forming bone matter. In this study, the pathogenicity of C. acnes in bone repair context was investigated. Materials and Methods. Human bone marrow derived MSCs were challenged with C. acnes clinical strains harvested from non-infected bone site (Cb). The behaviour of Cb strain was compared to C. acnes took from orthopaedic implant-associated infection (Ci). The infective capabilities of both strains was determined following gentamicin-based antibiotic protection assay. The morphology and ultrastructural analysis of infected MSCs was performed respectively through CLSM pictures of Phalloidin. ®. stained MSCs cytoskeleton and DAPI labelled Cb, and transmission and scanning electron microscopies. The virulence of intracellular Ci and Cb (Ci-MSCs and Cb-MSCs) was investigated by biofilm formation on non-living bone materials; and the immunomodulatory response of infected MSCs was investigated (PGE-2 and IDO secretion detected by ELISA). Bone cells (osteoblasts and PMA differentiated macrophages) were then challenged with Cb-MSCs and Ci-MSCs. Intracellular accumulation of ROS within infected macrophages was assessed by flow cytometry after 2 h of infection and the catalase production by Cb-MSC and Ci-MSC was evaluated. Statistical analyses were performed using Mann & Whitney test. Results. Following MSCs infection by C. acnes, the rate of viable bacteria inside MSCs was about 4% and 6% for Cb and Ci, respectively. Cb showed however a lower invasiveness in comparison to Ci (0.6-fold, p=0.01), confirming the higher pathogenicity of Ci. The ultrastructural and morphology analysis of infected MSCs confirmed the presence of bacteria free in MSCs cytoplasm, localized between F-actin fibers of MSCs, which preserved their elongated morphology. Considering the high level of secreted immunomodulatory mediators (PGE-2 and IDO), our results suggest that Cb-infected MSCs could promote a transition of macrophages from a primarily pro-inflammatory M1 to a more anti-inflammatory M2 phenotype. In comparison with Cb, Cb-MSCs increased significantly the formation of biofilm on TA6V and PEEK but reduced the biofilm formation on 316L SS. Ci-MSCs showed a significant increase in biofilm formation on PEEK vs Ci, while no difference in biofilm formation was noticed on TA6V and 316L SS. Regarding the ability of MSCs bacteria to infect osteoblasts, our results showed a higher infective capabilities of Cb-MSCs versus Cb (>2-fold, p=0.02), while no difference was noticed between Ci and Ci-MSCs. Along with an increase in catalase production by Cb-MSCs, we noticed its higher persistence to macrophage degradation. Conclusions. Taken together, our results demonstrate a shift in commensal Cb to pathogenic following infection. Indeed, Cb- MSCs acquires features that (i) increase biofilm formation on orthopedic based materials, (ii) increase the osteoblast infection and (iii) develop resistance to the macrophage degradation, through the increase of catalase production. Overall, these results showed a direct impact of C. acnes on bone marrow derived MSCs, providing new insights into the development of C. acnes during implant-associated infections

Background: In recent years there has been an increase in the insertion of prosthetic devices in orthopaedics. In spite of improvements in surgical techniques and antibiotic prophylaxis, the absolute number of infectious complications is high. Infections have a negative impact in patient’s quality of life and have high costs of management. Patients and Methods: Retrospective analysis of diagnosis, aetiology, and therapy of prosthetic devices infections observed from 1985 to 1999 in the operative unit for diagnosis and treatment of Infections in orthopaedics of Ospedale S. Corona- Pietra Ligure (SV). Results: During the study period, 251 patients with infected hip prosthesis and 133 with infected knee prosthesis had been treated. Diagnosis of infection was made by means of clinical features supported by x-ray, MRI, CT scan, ultrasonography and radio-nuclide scan. Aetiology was established by microbiological culture and histology. The majority of cases were single agent infections due to Gram-positives, especially S. aureus and S epidermidis, isolated in 41% of hip and 53% of knee prosthesis infection, while P. aeruginosa represented the most frequently isolated Gram-negative (3% in hip prosthesis and 10.6 % in kne prosthesis). Polymicrobial infections (with constant presence of S. aureus and/or S. epidermidis) accounted for 8% of hip and 7% of knee prosthesis infections. Treatment was represented by prolonged antibiotic administration (at least 8 weeks) associated with surgical debridment inacute infections, and two-stage exchange in chronic infections. In 23 hip infections in patients in poor clinical conditions or in suspected persistence of latent infection a new prosthesis was not replaced and Girdlestone’s hip arthroplasty was performed. Conclusions: Gram-positives are the main cause of orthopaedic infections but Gram-negatives, especially P aeruginosa, are often isolated. The treatment must necessarily be combined: antibiotic therapy and surgical treatment. Only in presence of optimal conditions a new prosthesis can be replaced

Purpose: With the growing risk of nosocomial infections, one might expect to see a reinforcement of septic isolation wards in orthopaedics and traumatology units. The question is however being revisited because of several factors. 1st: General Orthopaedics Units are practically the only hospital units caring for a minority of septic patients with often resistant germs and a majority of non-septic patients in the same setting. 2nd: The growing number of single-patient rooms procures confidence (whether justified or not). 3rd: Hygiene specialists are particularly wary of occult carriers of resistant bacteria and apply a single set of protective measures for all patients. 4th: Economic performance is given priority. Material and methods: We studied 1) the current situation in Orthopaedic units in University Hospitals in France and 2) the statistics from the Besançon University Hospital Hygiene Unit and from data in the literature. Results: 1) Interrogation of the 71 University Orthopaedics Units in France revealed that: 11 units have strict isolation wards; 40 have incomplete isolation wards; 20 make no distinction between septic and non-septic patients. 2) According to the Hygiene Unit statistics, the epidemiological load of S. aureus meti-R (SAMR), strains often implicated in orthopaedic infection, is much higher in the University Hospital polyvalent wards than in the Orthopaedic septic ward. Contamination between septic patients is low. Furthermore, hand-borne and airborne contamination are not controlled in wards other than septic wards. Data in the literature are not in agreement concerning this new trend in prevention by isolation. Discussion: a) One argument retained by all is that septic wards have an advantage in terms of efficacy and concentration of preventive measures. b) The growing workload in mixed units hinders strict application of preventive measures. c) A large number of temporary personnel (trainees, temporary employees, personnel untrained in sepsis prevention) are present in polyvalent units. d) Standardisation of preventive measures leads to an average level of prevention which lengthens the duration of care for non-septic patients and simplifies care for septic patients. e) The financial argument is impertinent compared with the consequences of contamination. Furthermore, a departmental structure would allow common use of the septic ward. Conclusion : Septic isolation wards (or a septic department) should be preserved. The orthopaedic surgeon, as a responsible actor in the fight against nosocomial infections, should in concert with the consulting hygienist, oppose purely administrative decisions

Aim. Current guidelines for the diagnosis of prosthetic joint infection (PJI) recommend collecting 4–5 independent tissue specimens, with isolation of indistinguishable organisms from two or more specimens. The same principle has been applied to other orthopaedic device-related infections (DRI) including fracture-related infections. However there are few published data validating this approach in DRI other than PJI. We evaluated the performance of different diagnostic cutoffs and varying numbers of tissue specimens for microbiological sampling in fracture-related infections. Method. We used standard protocols for tissue sample collection and laboratory processing, and a standard clinical definition of fracture-related infection. We explored how tissue culture sensitivity and specificity varied with the number of tissue specimens obtained; and with the number of specimens from which an identical isolate was required (diagnostic cutoff). To model the effect of the number of specimens taken we randomly sampled n specimens from those obtained at each procedure, excluding procedures from which less than n specimens were collected, and calculated sensitivity and specificity based on this sample. For each value of n we repeated this process 100 times to estimate the mean sensitivity and specificity for n specimens. Results. We analysed data for 246 cases of suspected fracture-related infection. 77 (31%) met the clinical definition of infection. A median of 4 independent tissue samples were obtained from each procedure (IQR 4–5). Culture sensitivity was highest and specificity lowest using a diagnostic cutoff of 1 specimen for isolation of an organism; specificity increased at the expense of sensitivity with diagnostic cutoffs of 2 or 3 specimens. Culture sensitivity increased as the number of tissue specimens obtained increased from 1 to 4. Although there was a corresponding decline in specificity with increasing numbers of tissue specimens obtained, this was negligible when a diagnostic cutoff of 2 or 3 specimens with identical organisms was used. Using a cutoff of 2 specimens with identical organisms, obtaining 4 specimens gave a sensitivity of 68% (55–78%) and a specificity of 95% (86–99%). Small numbers prevented meaningful analysis of the diagnostic performance of five or more specimens. Conclusions. These data are analogous to findings in prosthetic joint infections, and suggest similar principles may be applied to tissue sampling and culture interpretation in other orthopaedic DRI including fracture-related infection. A larger study is underway to evaluate the performance of greater numbers of tissue specimens

Aim. Cutibacterium acnes, a skin commensal, is responsible for 5–10% of prosthetic joint infections (PJI). All current microbiological definitions of PJI require two or more identical commensal isolates to be recovered from the same procedure to diagnose PJI and rule out contamination. Unlike coagulase negative staphylococci, C.acnes shows a highly stereotypical susceptibility profile making impossible to phenotypically assess the clonal relationship of isolates. In order to determine the clonal relationship of multiple C.acnes isolates recovered from arthroplasty revisions, we analyzed by multi-locus sequence typing (MLST) C.acnes isolates grown from orthopedic device-related infections (ODRI) in a reference center for bone and joint infection. Methods. Laboratory records from January 2009 to January 2014 were searched for monomicrobial C.acnes ODRI with growth of C. acnes in at least 2 intraoperative and/or preoperative samples. Clinical, biological and demographic information was collected from hospital charts. All corresponding isolates biobanked in cryovials (−80°C) were subcultured on anaerobic blood agar, and identification confirmed by MALDI-TOF-MS. C.acnes isolates were typed using the MLST scheme described by Lomholt et al. Plasmatic pre-operative C-reactive protein (CRP) levels were determined using DimensionEXL (Siemens). A threshold of 10 mg/L was used to determine serologically positive ODRIs from negatives. Results. Over a 5-year period, 37 cases of monomicrobial C.acnes ODRI were diagnosed in our center. Among these 37 cases, 113/153 C.acnes isolates were cryopreserved. 110/113, corresponding to 36/37 cases, were typed by MLST: 14/36 (39%) ODRI cases were found to feature isolates belonging to two or more different STs and were qualified to be heteroclonal whereas 22/36 (61%) of ODRI cases were found to feature isolates belonging to the same ST and were qualified to be homoclonal. Homoclonal infections were significantly more likely to have elevated CRP levels compared to heteroclonal cases (p=0.0011, Fisher test). Patients with only two positive intraoperative samples had significantly lower CRP values than patients with three or more positive intraoperative samples (12,7mg/L vs 67mg/L; p=0,01, homoscedastic two-tailed Student's t test). Conclusions. This study suggests that what is classified microbiologically as C.acnes ODRIs comprises: i) true homoclonal infections eliciting an inflammatory response, ii) heteroclonal infections lacking inflammatory response where C.acnes could be an innocent bystander and iii) false positives where no strain achieves true microbiological significance. Our study shows that a stricter threshold of 3 intraoperative positive samples could be more adequate than 2. These results reinforces the need for a more specific definition of C.acnes ODRI

Aim. To assess the clinical characteristics, diagnostic tests and treatment strategies in orthopedic implant-associated infections (OIAI) caused by Cutibacterium spp. Method. We retrospectively included consecutive patients with OIAI caused by Cutibacterium spp. treated at our institution from January 2012 to January 2017. OIAI was diagnosed when: (i) macroscopic purulence, sinus tract or exposed implant was present; (ii) acute inflammation in peri-implant-tissue was documented; (iii) Cutibacterium spp. grew in joint aspirate, ≥2 intraoperative peri-implant tissue samples or in sonication fluid of the removed implant (>50 CFU/ml). Results. Of 67 patients with Cutibacterium OIAI, 42 (63%) had an infected joint prosthesis (21 hip, 12 shoulder, 9 knee) and 25 (37%) an infected fixation device (10 spinal hardware, 11 osteosynthesis, 2 anchorages after rotator cuff reparation, 2 cruciate ligament grafts). 53 (84%) presented with a delayed (3–24 months) or late (>24months) infection. 62 infections were caused by C. acnes and 5 by C. avidum, all being susceptible to levofloxacin and rifampin. Among non-culture-based diagnostic tests, tissue histology had the highest sensitivity (68%), followed by increased synovial fluid leukocyte count/differential (59%). Of culture-based tests, sonication fluid culture showed the highest sensitivity (83%), followed by tissue culture (71%) and synovial fluid culture (61%). Culture positivity rates of synovial fluid, peri-implant tissue and sonication fluid were 20%, 41% and 40%, respectively, after 7 days of incubation and 58%, 74% and 78%, respectively, after 14 days. Most patients were treated with one-stage (24%) or two-stage (55%) implant exchange of the implant. The majority of patients received oral levofloxacin and rifampin for 6–12 weeks. Conclusions. Cutibacterium spp. affected various types of orthopaedic implants in different anatomic locations (lower and upper limbs and spine). Conventional diagnostic tests showed limited sensitivity of Cutibacterium OIAI and can be easily missed when cultures are incubated less than 10–14 days. All Cutibacterium isolates were susceptible to levofloxacin and rifampin

Aim. Staphylococcus epidermidis has emerged as an important opportunistic pathogen causing orthopedic device-related infections (ODRIs). In this prospective clinical and laboratory study, we have investigated the association of genome variation and phenotypic features of the infecting S. epidermidis isolate with the clinical outcome of the infected patient. Method. One hundred and four invasive S. epidermidis isolates were prospectively collected from patients with ODRI. Upon patient entry into the study, surgical parameters such as type of implant; open or closed fracture were documented. Personal characteristics were also documented and included: gender; age; body mass index (BMI); smoker/non-smoker; overall medical condition (Charlson comorbidity index); and chronic immunosuppressive conditions. Any revision surgeries involving the site of interest and all isolated pathogens were recorded throughout the course of treatment and follow-up. The clinical outcome after treatment was measured with a mean follow-up period (FUP) of 26 months, and each patient was then considered to have been “cured” or “not cured”. The isolates were tested for their antibiotic susceptibility and ability to form biofilm. Whole genome sequencing was performed on all isolates and genomic variation was related to features associated with “cured” and “not cured”. Results. Strong biofilm formation and resistance to aminoglycoside antibiotics were associated with a “not cured” treatment outcome (p = 0.031 and p < 0.001, respectively). Isolates within the “not cured” group were more prevalent in the phylogenetic clade B compared to the predominant clade A (p = 0.08). Based on a gene-by-gene analysis, some accessory genes were more prevalent in isolates from patients that were “not cured”. These included: the biofilm-associated bhp gene; the antiseptic resistance qacA gene, the cassette chromosome recombinase encoding genes ccrA and ccrB and IS256-like transposase. Conclusions. The novelty of this study was that all S. epidermidis isolates were whole genome sequenced in order to screen for features associated with poor clinical outcome. Multiple factors were found to have an influence on poorer patient clinical outcome, such as multiple revisions surgeries, biofilm formation and antibiotic resistance to aminoglycosides emphasizing the multifactorial pathogenesis of ODRI. Furthermore, poor treatment outcome was associated with a small number of mobile elements that could potentially be used as prognostic markers for ODRI treatment outcome

Aim. Current standard of care in the management of bone and joint infection commonly includes a 4–6 week course of intravenous (IV) antibiotics but there is little evidence to suggest that oral antibiotic therapy results in worse outcomes. The primary objective was to determine whether oral antibiotics are non-inferior to IV antibiotics in this setting. Method. This was a parallel group, randomised (1:1), open label, non-inferiority trial across twenty-six NHS hospitals in the United Kingdom. Eligible patients were adults with a clinical diagnosis of bone, joint or orthopaedic metalware-associated infection who would ordinarily receive at least six weeks of antibiotics and who had received ≤7 days of IV therapy from the date of definitive surgery (or the start of planned curative treatment in patients managed non-operatively). Participants were randomised to receive either oral or IV antibiotics for the first 6 weeks of therapy. Follow-on oral therapy was permitted in either arm. The primary outcome was the proportion of participants experiencing definitive treatment failure within one year of randomisation. The non-inferiority margin was set at 7.5%. Results. Of 1054 participants randomised (527 to each arm) endpoint data were available for 1015 (96.30%). Definitive treatment failure was identified in 141/1015 (13.89%) participants, 74/506 (14.62%) of those randomised to IV therapy and 67/509 (13.16%) of those randomised to oral therapy. In the intention to treat analysis, the imputed risk difference (PO-IV) for definitive treatment failure was −1.38% (90% CI: −4.94, 2.19), thus meeting the non-inferiority criterion (i.e. the upper limit of 95%CI being <7.5%). A complete cases analysis, a per-protocol analysis and sensitivity analyses for missing data confirmed this result. With the exception of intravenous catheter complications, there was no significant difference between the two arms in the incidence of serious adverse events (SAEs). Health economic analysis suggests that the non-surgical treatment costs over one year for patients randomised to oral therapy were approximately £2,700 less than those of IV therapy. Conclusions. Oral antibiotic therapy is non-inferior to IV therapy when used during the first six weeks in the treatment for bone and joint infection, as assessed by definitive treatment failure within one year of randomisation. These findings challenge the current standard of care and provide an opportunity to realise significant benefits for patients, antimicrobial stewardship and the health economy. Funding. The OVIVA study was funded by the National Institute for Health Research Health Technology Assessment programme (Project number 11/36/29)

Aim. The treatment of chronic orthopedic device-related infection (ODRI) often requires multiple surgeries and prolonged antibiotic therapy. In a two-stage exchange procedure, the treatment protocol includes device removal and placement of an antibiotic-loaded bone cement spacer to achieve high local antibiotic concentrations. At the second stage, further surgery is required to remove the spacer and replace it with the definitive device. We have recently developed a thermo-responsive hyaluronan hydrogel (THH) that may be loaded with antibiotics and used as delivery system. Since the material is bio-resorbable, it does not require surgical removal and may therefore be suitable for use as treatment strategy in a single-stage exchange. This aim of this study was to evaluate gentamicin sulphate (Genta)-loaded THH (THH-Genta) for treating a chronic Staphylococcus aureus ODRI in sheep using a single-stage procedure. Methods. Twelve Swiss-alpine sheep received an IM tibia nail and an inoculation of a gentamicin-sensitive clinical strain of Staphylococcus aureus. After letting a chronic infection develop for 8 weeks, a revision procedure was performed: the implant was removed, the IM canal debrided and biopsies were taken for culture. The IM canal was then filled with 25ml THH-Genta (1% Genta) or left empty (control group) prior to the implantation of a sterile nail. An ultrafiltration probe was placed within the IM cavity to collect extracellular fluid and determine local antibiotic levels for 10 days. Both groups received systemic amoxicillin and clavulanic acid for 2 weeks, followed by 2 weeks without treatment for antibiotic washout. At euthanasia, IM nail, bone marrow, bone and tissue samples were harvested for quantitative bacteriology. Results. All sheep were infected at revision surgery as confirmed by cultures of biopsies and sonication of the IM nail. Local Genta concentrations ranged on average from 830µg/ml postoperatively to below 5µg/ml after 8 days. At euthanasia, S. aureus was detected in 5/5 IM nails, 5/5 bone marrow samples, and 8/25 superficial soft tissue samples in the control group (one control sheep was excluded for having a superinfection). In the THH-Genta group, S. aureus was cultured from 0/6 IM nails, 1/6 bone marrow samples, and 1/30 superficial soft tissue samples. Conclusions. The THH showed a Genta release pattern that started with high local concentrations and decreased to low concentrations within 10 days. Local Genta delivery by THH combined with systemic antibiotics significantly reduced infection rates whereas systemic therapy alone was unable to eradicate infection in any animal

Aim. Propionibacterium acnes is an emerging pathogen especially in orthopedic implant infection. Interestingly, we previously reported a difference in the distribution of the clades involved in spine versus hip or knee prosthetic infection. To date, no study has previously explored the direct impact and close relationship of P. acnes on bone cells according to their own genetic background. The aim of this study was to investigate this interaction of P. acnes clinical strains involved in spine material infections, arthroplasty infections and acne lesions with bone cells. Method. From a large collection of 88 P. acnes clinical isolates collected between January 2003 and December 2014, a subset of 11 isolates was studied. Four isolates were recovered from spine infections, two from prosthetic infections (knee and hip), three from acne lesions and two reference strains (ATCC11827 and ATCC6919). Implant-associated infections were confirmed according to Infectious Diseases Society of America guidelines for bone and joint infections. Multi-Locus Sequence Typing (MLST) was carried out on all isolates as described by Lomholt et al. PLoS ONE 2010. Bacterial internalization experiments with MG63 osteosarcoma cells were adapted from Crémet et al. Pathog Dis 2015. Results. Among the nine clinical isolates, three isolates belonged to clonal complexes (CCs) 18; three to CC28 and three to CC36. ATCC isolates belonged to CC18. Bacterial internalization experiments revealed that CC36 P. acnes strains were less invasive than CC18 and CC28 P. acnes strains towards osteoblasts (mean percentage of internalized bacteria (< 0.01% for the CC36 P. acnes strains versus more than 1% for the CC18 and CC28 P. acnes strains). Surprisingly, the ATCC11827 CC18 P. acnes strain exhibited invasiveness similar to CC36 isolates. Conclusions. Evasion mechanism observed for CC36 P. acnes isolates could allow this clade to leave the site of infection, disseminate into deeper tissue layers and beget arthroplasty infection. Inside the deeper tissue, close to the material, the local immune defect fosters the low-grade infections observed with P. acnes clinical strains. On the another hand, for CC18 et CC28 clades, mostly involved in spine infection, the internalization process observed could allow these clades to escape from the numerous immune cells located under the skin and generate an infection locally, favored by the spine instrumentation close to the skin, especially during long spine surgeries

Propionibacterium acnes is an emerging pathogen especially in orthopedic implant infection. Aim of this study was to investigate P. acnes phylogeny and to screen for virulence factors among a large collection of clinical isolates involved in spine material infections, arthroplasty infections and acne lesions. 88 P. acnes clinical isolates were collected between January 2003 and December 2014 at Nantes University Hospital (France). Fifty-eight isolates came from spine infections, 14 from prosthetic infections (knee, hip or shoulder), 14 from acne lesions and two reference strains (ATCC11827 and ATCC6919). Implant associated infections were confirmed using Infectious Diseases Society of America criteria for bone and joint infections. Phylotypes and Multi-Locus Sequence Typing (MLST) was carried out on all isolates as described by Lomholt et al. All isolates were tested by established PCR-based assays for 21 putative virulence factor genes characteristic of P. acnes. MLST analysis revealed an association between clonal complexes (CCs) and origin of P. acnes isolates (p = 0,027). Regarding CCs distribution between different origins, CC36 and phylotype II P. acnes isolates are more frequently observed in prosthetic joint infections. On the other hand, CC18 (IA) and CC28 (IB) P. acnes isolates are more frequently involved in spine infections and acne lesions. Among all virulence factors screened, hyaluronate lyase gene was only present in CC36 and phylotype II P acnes isolates. Other virulence factors were present in all isolates, whatever their origin or CC. Regarding molecular typing results, P. acnes involved in spine infections seem to have a skin origin (same CC as isolates from acne lesion). Interestingly, the origin of prosthetic joint infection isolates seems different and they all carry one more virulence factor. Hyaluronate lyase (Hyl) is a major surface protein of P. acnes with potential antigenetically variable properties that might be essential for P. acnes virulence. Increased tissue permeability caused by the action of hyaluronidase on the extracellular matrix appears to play a role in wound infections, pneumonia, and other sepsis such as bacteremia and meningitis. It could be also take a prominent part in P. acnes prosthetic joint infection pathogenesis

The treatment of orthopedic implant infections is often difficult and complex, although the chances of successful treatment with a properly selected diagnostic, surgical and antibiotic treatment protocol have recently increased significantly. Surgical treatment is a key factor in the treatment of infections of orthopedic implants, and any errors in this respect often lead to worse clinical outcomes. Surgical errors. The most important and frequent surgical errors include:. - conservative treatment of periprosthetic infections with antibiotics alone: successful treatment requires adequate surgical procedure combined with long-term antimicrobial Th that is active against biofilm microorganism. Without adequate surgical procedure just the suppression of symptoms is usually achieved, rather than eradication of the infection. - delayed surgical revision: in acute infections, early surgical intervention plays a critical role, especially by patients where retention of the prosthesis is expected. Early evacuation of postop haemathoma after primary or revision surgery is important in order to prevent the possibility of infection. It is important to take into consideration, that a postop apparently superficial surgical site infection may be indicative of deeper infection involoving the implant. - insufficient debridement during surgical revision: thorough and extensive debridement is the most critical predictor of success (removal of the haemathoma, abscess formations, fibrous membranes, sinus tracts, devitalized bone and soft tissue, removal of all cement, cement restrictors, foreign and prosthetic material; eventual exchange of modular components and liners). Finally meticulous irrigation of the op region is obligatory. - inadequate intraoperative sampling for bacteriological and histological analysis: tissue samples from the areas with the most florid inflammatory changes have to be taken and sent for bacteriological and histological examination (3–6 samples). Removed implants or parts of them have to be sent to sonication. Swab cultures have low sensitivity and should be avoided. - the importance of selecting the appropriate surgical strategy for the individual patient cannot be overemphasized: not having, following and treating patients with PJI accordingly to an algorithm that is proven and successful one usually leads to unsuccessful clinical results. We present illustrative cases with each common surcical error combined with proper solution. Treatment of PJI is a demanding procedure, the goal is a long-term pain-free functional joint, that can be achieved by eradication of the infection. For a successful clinical outcome an appropriate diagnostic, surgical and antimicrobial procedure for the individual patient has to be selected

Introduction. In 2011 the Scottish Government published national MRSA screening requirements. A comparison of Orthopaedic and ENT elective surgery intended to juxtapose a specialty known to take MRSA screening seriously with one that has little clinical concern with regards MRSA infection. ENT surgery parallels Orthopaedics in using implants and there potentially being MRSA colonisation at or close to the site of surgery. In Orthopaedics MRSA infection is infrequent, but implant infection with antibiotic resistant bacteria has a particularly poor prognosis. In ENT MRSA infection is rare and colonisation does not influence patient care. Aims. An evaluation of MRSA screening practice for elective Orthopaedics and ENT surgery at Gartnavel General Hospital with regards strategy and implementation. Method. Review of 342 consecutive elective ENT patients and 325 Orthopaedic patients attending for inpatient or day case surgery. The reference standards were the regional and national guidelines on MRSA screening. Results. Overall screening rates were 145 (42%) of 342 ENT patients and 270 (83%) of 326 Orthopaedic patients. 100% of Orthopaedic patients admitted (154) were screened, in compliance with both regional and national policy. 91 (70%) of 130 ENT patients admitted were screened for MRSA, and no risk assessment was carried out, which was not in compliance with either regional or national policy. Discussion. Orthopaedic surgery has an established and reliable practice of screening elective inpatient cases, and when identified MRSA colonisation results in a change in patient management. ENT surgery should have established a similar practice according to existing local guidelines. The Government consider ENT a lower risk speciality for MRSA, but still require as a minimum a documented MRSA risk assessment process

The February 2024 Research Roundup³⁶⁰ looks at: If you use a surgical helmet, you should seal your gown-glove interface; The use of iodophor-impregnated drapes in patients with iodine-related allergies: a case series and review of the literature; Location of the ovaries in children and efficacy of gonadal shielding in hip and pelvis radiography; Prehospital tranexamic acid administration does not improve outcomes in severe trauma patients; Silver-coated distal femur megaprosthesis in chronic infections with severe bone loss: a multicentre case series.

Intramedullary nailing (IMN) has been frequently indicated to treat long bone open and closed fractures, but infection following internal fixation may have devastating consequences, with higher costs. Treatment of intramedullary nail-associated infections (IMNI) is challenging and based upon surgery and adequate antibiotic administration, which requires the correct identification of causative microorganisms. However, there have been difficulties for the microbial diagnosis of IMNI, as the peri-prosthetic tissue cultures may show no microbial growth, particularly in patients with previous use of antibiotics. Sonication have shown higher sensitivity and specificity for microbial identification on a variety of orthopedic implant-associated infections. Aim: To compare clinical and microbiological results and sensitivity for the pathogen identification obtained by conventional peri-implant tissue culture samples with culture of samples obtained by sonication of explanted IMN implants, among patients presenting IMNI of long bones. Methods: Longitudinal prospective cohort study performed at a tertiary public hospital, ongoing since August 2011. We analyzed all patients with indication for IMN implant removal, and orthopedic-implant associated infections was defined according to previous publications addressing osteosynthesis-associated infections (Yano 2014). Minimal of 2 samples from the peri-implant tissue were taken and sent under sterile conditions to the laboratory for culture. Statistical analysis was performed McNemar's test for related proportions. Results: We included 26 patients presenting clinical signs of IMNI, of which tissue and sonication cultures were performed for 26 (100%) and 20 (77%) patients, respectively. Among them, 88% were male, with mean age was 35.9 years (range, 19–59 yo). Causes of trauma were mainly motorcycle crashes accounting 54% of accidents; tibia and fibula were affected in 65% and 27%, respectively. Gustilo open fracture classification was grade II (35%) and IIIA (35%). First stage management with external fixation for fracture stabilization was performed in 75% of trauma patients. Sensitivity of peri-prosthetic tissue culture and sonication was 80.7% (21/26), and 95% (19/20) (p< 0.05), respectively. Only one infected patient presented negative tissue and fluid cultures. Gram-positive cocci were isolated in 75% and 79% in tissue and sonication fluid cultures, respectively. Staphylococcus aureus, coagulase-negative staphylococci, Enterococcus sp., were isolated from tissue and sonication culture in 43.5% and 36.3%, 8.7% and 22.7%, 13% and 13.7%, respectively. Polymicrobial infection was diagnosed in 3.8% (1/26) and 15.8% (3/19), patients by tissue and sonication fluid cultures (p< 0,01), respectively. Conclusion: Sonication of retrieved infected intramedullary nails has the potential for improving the microbiological diagnosis of IMNI

Aims

The duration of systemic antibiotic treatment following first-stage revision surgery for periprosthetic joint infection (PJI) after total hip arthroplasty (THA) is contentious. Our philosophy is to perform an aggressive debridement, and to use a high local concentration of targeted antibiotics in cement beads and systemic prophylactic antibiotics alone. The aim of this study was to assess the success of this philosophy in the management of PJI of the hip using our two-stage protocol.

Aims

This study aimed to explore the role of small colony variants (SCVs) of Staphylococcus aureus in intraosseous invasion and colonization in patients with periprosthetic joint infection (PJI).