Frozen section is a recognised technique to assist in the diagnosis of infection and there are standards for reporting. Our aim of this review was to assess the value of frozen section in the diagnosis of infection, as well as other variables. We performed a retrospective review of all frozen sections for suspected infection in 2016. Patient demographics, histological and microbiological investigations, laboratory and bedside tests were recorded and analysed using statistical software. 46 patients had 55 frozen sections; the majority were for lower limb arthroplasty. No sections were reported as polymorphonuclear neutrophils per high-power field. Three sections showed signs of infection and one without evidence had positive cultures. One uncertain section did not grow organisms. 10 patients had two-stage procedures, four of these were intended to be determined by frozen section but only two had evidence of infection on analysis. Evidence of infection on frozen section does correlate with microbiological growth but does not relate to intention to stage procedures in half of patients. The effect of new tests such as Synovasure is highlighted by this review. Frozen section analysis is reported subjectively but is a good predictor of infection. Clinical assessment is accurate in diagnosing infection. Histological, microbiological and additional investigations should be considered in relation to their cost-effectiveness

Aim. To assess the effectiveness of role of frozen section in revision arthroplasty. Method. 21 patients with infected hip arthroplasties were operated in the form of one or two-staged revision hip arthroplasties. A frozen section was obtained intra-operatively and >5 PMN's/ HPF was considered as a positive indicator of infection. Fig 1 llustrating frozen section image. If the frozen section was reported negative (≤5 PMN's/HPF), the revision prosthesis was implanted after a thorough debridement and a wash. If the frozen section was reported as positive, after the debridement a non-articulating antibiotic loaded cement spacer was implanted for 8 weeks, supplemented with 3 weeks of intravenous antibiotics and 3 weeks of oral antibiotics. This was followed by an antibiotic free interval of 2 weeks. The patient was taken up for a revision surgery once the frozen section study was negative (≤5 PMN's/HPF). The patients were followed up for minimum of 1 year to a maximum of 2 years after the revision for any evidence of infection (assessed clinically and serologically, radiologically). Results. 15 patients had a positive frozen section (>5PMN's/HPF) in the first stage and were treated with prosthesis removal and cement spacer insertion for 8 weeks. In the 2nd stage, out of 15 patients, 14 underwent revision arthroplasty, while 1 patient underwent reapplication of the cement spacer. As per the follow up of ESR & CRP values, clinically and radiologically no patients had any evidence of infection. The average follow up was 17.04 months (range 12–24 months). 1 patient had persistently raised ESR (34mm/hr) which may be attributable to other causes Frozen section analysis of PMN's per high power field had 100% specificity in our patients in detecting periprosthetic joint infection. Conclusions. Intraoperative frozen section study is a reliable indicator in predicting a diagnosis of PJI with good accuracy in ruling out this diagnosis. Frozen section study should thus be considered a relevant part of the challenging diagnostic work-up for patients undergoing revision hip arthroplasty

Injection before total knee arthroplasty(TKA) is the one of the postoprative risk factors after TKA and Infection after TKA can result in disastrous consequences. When the duration between injection and TKA is longer than 6 months, the risk is no longer elevated. Evaluation of synovial WBC number in frozen section slide is needed to check the presence of infection in revision total knee arthroplasty. Currently many patients have a history of multiple intraarticular injection before the primary TKA. Purpose of this study is to evaluate the synovial WBC findings in primary TKA and compare between injection group and no injection group. Materials and Methods. The synovial specimen(suprapatella pouch and posterior capsule) of 68 primary total knee arthroplasty were evaluated by the pathologist and reported the number of the WBC in frozen section /5 separate high power fields(HPF) (500x).. Injection group were 37 cases and non -injection group were 31 cases. Preoperative CRP and ESR were recorded and followe-up duration was more than 2 years. Joint fluid was sent to be cultured and analysed. Results. WBC count in frozen section shoed was average 4 WBCs/HPF (range < 0∼ 25) in both specimen and the suprapatella specimen was 3 WBCs/HPW (range 0∼25) and posterior capsule specimen was 1 WBCs/HPF(range 0∼14). The WBC count of injection group was 8 (range, 0∼25) and that of no injection group was 1.2 cells (range 0∼12) (p<0.05). The WBC counts in joint fluid was average 240 cells/ml (range. 1∼300) in non injection group and 643 cells/ml(range, 50∼1000) (p<0.05). The duration from the intraarticular injection to index surgery was 9 months(range, 6 weeks∼ 7 momths). The number of injection and duration bwtween injection and operationto has no significant correlation with the WBC counts. Eight percentage of specimen showed more than 10 WBCs in injection group and these patients have been not infected after more than 24 moths after TKA. Conclusion. The WBC count of the synovium in priamry TKA with injection history for degenerative osteoarthritis is variable and we could not recommend the routine frozen section analysis in primary TKA who have a history of intraarticualr injection

The purpose of this study was to evaluate the accuracy of the sonication fluid cultures (SFC) for the diagnosis of prosthetic joint infection and compare it with frozen section and periprosthetic tissue cultures. 108 patients underwent revision or explantation procedure for any reason. Frozen sections of intraoperative specimen were analized and multiple periprosthetic samples (at least 5) were collected and cultured. All explanted prosthesis components were subject to sonication and cultured. All cultures were incubated for 14 days. PJI was diagnosed in 52 patients (48%). Sonication achieved the highest sensivity with 95% and specificity of 98%. Frozen section showed low sensivity (44%) and specificity (80%) and periprosthetic tissue cultures showed sensivity of 75% and specificity of 98%. Sonication fluid culture is a cheap, easy, accurate and sensitive diagnostic method and helps to detect about 30% more PJI compared to frozen section and 16% more compared to periprosthetic tissue cultures. It also detect about 25% more pathogens than periprosthetic tissue cultures

Infection in arthroplasty surgery is a major complication leading long antibiotic courses and frequently requiring repeated operations to eradicate or suppress. Therefore in the situation of revision surgery on prosthesis that are possibly already infected a clear identification of possible infection is required. Previously frozen section samples have been used in Derriford Hospital in conjunction with clinical presentation and other investigations to aid in diagnosis and tailor management, however recent studies have suggested that this may not be as effective as previously thought. Kanner et al. (2008) suggested a sensitivity of 29% and positive predictive value of 40%. This retrospective audit reviewed the cases between March 2007 and May 2012, identifying 220 cases of revision surgery where infection was suspected and frozen sections analysis was performed. Results where then compared to paraffin and cultured samples if taken. A notes review was performed to demonstrate if the operative technique (single or two stage) was in line with local guidelines for the results of the frozen section. Long term survival (longest follow up of 7 years) was assessed by need for revision surgery

Aim. Diagnosing low-grade periprosthetic joint infections (PJI) can be very challenging due to low-virulent microorganisms capable of forming biofilm. Clinical signs can be subtle and may be similar to those of aseptic failure. To minimize morbidity and mortality and to preserve quality of life, accurate diagnosis is essential. The aim of this study was to assess the performance of various diagnostic tests in diagnosing low-grade PJI. Methods. Patients undergoing revision surgery after total hip and knee arthroplasty were included in this retrospective cohort study. A standardized diagnostic workup was performed using the components of the 2021 European Bone and Joint Infection Society (EBJIS) definition of PJI. For statistical analyses, the respective test was excluded from the infection definition to eliminate incorporation bias. Receiver-operating-characteristic curves were used to calculate the diagnostic performance of each test, and their area-under-the-curves (AUC) were compared using the z-test. Results. 422 patients undergoing revision surgery after total hip and knee arthroplasty were included in this study. 208 cases (49.3%) were diagnosed as septic. Of those, 60 infections (28.8%) were defined as low-grade PJI (symptoms >4 weeks and caused by low-virulent microorganisms (e. g. coagulase-negative staphylococci, Cutibacterium spp., enterococci and Actinomyces)). Performances of the different test methods are listed in Table 1. Synovial fluid (SF) - WBC (white blood cell count) >3000G/L (0.902), SF - %PMN (percentage of polymorphonuclear neutrophils) > 65% (0.959), histology (0.948), and frozen section (0.925) showed the best AUCs. Conclusion. The confirmatory criteria according to the EBJIS definition showed almost ideal performances in ruling-in PJI (>99% specificity). Histology and synovial fluid cell count (SF-WBC and SF-%PMN) showed excellent accuracies for diagnosing low-grade PJI. However, a reduced immune reaction in these cases may necessitate lower cut-off values. Intraoperative frozen section may be valuable in cases with inconclusive preoperative diagnosis. For any tables or figures, please contact the authors directly

Introduction. In revision surgery, detection of periprosthetic joint infection is of prime importance. Valuable preoperative and intraoperative diagnostic tests and tools are necessary. The classical standard procedures are puncture and bacteriology examination, frozen section intraoperative and powerfield micro analysis. Since autumn 2014 a new device for detection of periprosthetic joint infection is available, named Synovasure. It is a fast test for the detection of Alpha defensing, which plays a major role in the antimicrobial defence and only occurs in inflammatory processes. „The alpha-defensin test is an immunoassay that measures the concentration of the alpha-defensin peptide in human synovialfluid. A-Defensin is an antimicrobial peptide that is secreted into the synovial fluid by human cells in response to pathogenic presence” (Deirmengian C et al., CORR 2014). Summarized, the evidence of Alpha defensin indicates infection. It is produced by CD Diagnostics (Wynnewood, PA, USA) and merchandised by Zimmer (Warsaw, IL, USA). We are using Synovasure in daily routine at our department since September 2014. The aim of this conducted study is to present our first clinical experience and to report our results of the first 54 cases. Material and Methods. At our department Synovasure is standardly used in hip and knee revision surgery. Additionally an intraoperative frozen section and a standard bacteriology were performed. The explanted endprosthesis were sent to examination by sonification in order to gain culture of the sonification fluid and were further examined by Multiplex PCR. A pathologist with more than 15 years of experience conducted the frozen section. The results of Synovasure were matched with all above examinations in order to describe specifity and sensitivity of it. Results. A negative Synovasure Test during surgery and a negative PCR were observed in 3 patients, however, the bacterial culture was positive (after 14 days of breeding) as well as the Multiplex PCR. One patient had a negative frozen section and a negative culture but a positive PCR. Another patient with a high CRP level, all clinical signs of infection and a positive Synovasure Test, had 6 negative cultures. This patient suffered from a Metallosis as well, due to a broken PE inlay of the TKA, which supports the previously stated that Metallosis may interfere this new tool. Unfortunately in this patient neither a frozen section nor a PCR are available. One patient, who had explanation due to infection, underwent reimplantation. During surgery the Synovasure Test and the frozen section were negative (Synovial Fluid), but postoperatively a positive culture and a positive histological report for infection were assessed. Furthermore, a total of 5 tests showed an application error and the test did not show any control line. Conclusion. In conclusion Synovasure helps to detect perprosthetic joint infection in an easy and fast manner. It is simple to integrate into daily routine, nevertheless all standardized examinations for infection need to be conducted

Introduction. The intra-operative diagnosis of Prosthetic Joint Infection (PJI) is a dilemma requiring intra-operative sampling of suspicious tissues for frozen section, deep tissue culture and histopathology to secure a diagnosis. Alfa defensin-1 testing has been introduced as a quick and reliable test for confirming or ruling out PJI. This study aims to assess its intra-operative reliability compared to the standard tests. Methods. Twenty patients who underwent revision hip and knee arthroplasty surgery were included. Patients joint aspirate was tested intra-operatively with the Synovasure kit, which takes approximately ten minutes for a result. Our standard protocol of collecting 5 deep tissue samples for culture and one sample for histopathology was followed. Results for Alfa defensin-1 test were then compared with final culture and histopathology results in all these patients. Results. Our results show an excellent correlation with the final deep tissue cultures and histopathology outcomes. Literature reports frozen section to have low (58–73%) sensitivity but high (96%) specificity. Conclusions. Alfa defensin-1 test is easy, quick and efficient; results were available immediately intra-operatively. Cryosection is time consuming with samples shipped to the reference laboratory at times resulting in intra-operative delays. In our practice Alfa defensin-1 test certainly will replace frozen section for intra-operative testing

Revision of infected TKA is one of the most challenging operation as the surgeon should achieve two goals, ie eradication of infection and restoration of function. For the eradication of infection, a minimum of two operations are needed in most of cases. First stage of revision is meticulous debridement and insertion of antibiotic loaded cement. During arthrotomy, thick fibrous and granulation tissues which is located in the suprapatella pouch, lateral site to the patella tendon and posterior joint space should be removed so as to get better exposure, to get rid of infection source and to get better functional result. During debridement, I use highly concentrated antibiotic saline (1 gm vancomycin in 10cc saline), for irrigation of the operation field. I also pack the opening of the medullary canal so as to prevent the debris from entering into the medullary canal. I use antibiotics with the ratio of 1:3. To reduce the dead space in the medullary canal I insert a dowel shaped antibiotic loaded cement spacer made from one pack of cement and fill the medullary canal. Thereafter two packs of cement are used to make a block to fill the gap between femur and tibia. The cement block should be large enough to cover the distal femur and proximal tibia so as not to cause bone defect and knee dislocation during walking. After first stage of operation, antibiotics are administered for 4∼8 wks until the CRP levels become normalized and clinical findings show no sign of infection. The 2. nd. stage of operation is planned when clinical and laboratory signs of infection subside. The decision whether to reimplant the prosthesis or not is based on the operation findings and polymorphonuclear cell count on frozen section. However operation findings are considered more important than the frozen section results for reimplantation. If operative findings are clean, I do reimplanation even though the polymorphonuclear cell count is more than 5 on high power field(hpf) on frozen section. I have adopted numbering system to take specimen. Number 1 is specimen from suprapatella pouch, No 2 is that from gap between the femur and tibia, No 3 is that from femoral intramedullary canal, No 4 is that from tibial intramedullary canal, and No 5 is that from most unhealthy site. In a retrospective analysis of 16 cases which received reimplantation despite of the prescence of more than five polymorphonuclear cells on intra-operative frozen sections, none of the cases had recurrence of infection at a final follow up of 2 years. The femoral medullary canal was the most prevalent site for higher polymorphonuclear cell count. In conclusion, indication is the first step for successful reimplantion. Two stage revision is recommended and meticulous debridement is utmost important in first stage operation. Block type antibiotic loaded cement is sufficient for a good result. Clinical, laboratory and operative findings are more important than polymorphonuclear cell count on frozen section to decide reimplantation. I propose numbering system of the specimen site for frozen section, just as in tumor surgery

Peri-prosthetic joint infection (PJI) can be both a diagnostic and therapeutic challenge in shoulder arthroplasty, due to the indolent nature of the common infecting organisms. Proprionobacterium acnes (P. acnes) is the most common pathogen cultured in revision shoulder arthroplasty. It is a slow growing, anaerobic organism – requires longer incubation period (7–21 days). Coagulase-negative Staphylococcus species (CNSS) is also a common organism responsible for PJI. Established diagnostic tests for hip and knee PJI are often negative in the shoulder despite post-operative growth of intra-operative cultures. Pre-operative synovial aspiration often low volume due to indolent pathogens and successful aspiration is often reported to be 50% or less with Dilisio et al, JBJS 2014: reporting 16.7% sensitivity, 100% specificity. Variable culture length for P. acnes culture protocols are reported from 7–28 days with most groups recommending 14 days. From our research, we demonstrated time to culture growth was significantly shorter in probable true positive culture group (median, 5 vs. 9 days, p=0.002). Frozen section analysis may help intra-operative decision-making (one- vs. two-stage reimplantation) yet the reported sensitivity and specificity in shoulder arthroplasty is far less than in hip and knee arthroplasty. Synovial fluid biomarkers have been identified as part of the innate response to pathogens include pro-inflammatory cytokines and antimicrobial peptides. In a series of prospective studies of revision shoulder arthroplasty, synovial fluid analysis reported by Frangiamore et al, JBJS 2015: IL-6, Frangiamore et al, JSES 2015: α-defensin (Synovasure. TM. ), Frangiamore et al, AAOS 2015: Broader cytokine analysis it was demonstrated that these markers are much more predictive of infection than synovial fluid cultures, frozen section or serum markers

We investigated the role of Plasma Viscosity (PV), C-reactive protein (CRP) and Frozen Section (FS) in diagnosing prosthetic joint infection. We compared these results with microbiological diagnosis of infection of the tissue samples (three or more samples grown same organisms in culture). 53 patients, average age 67 years (37 – 89) underwent joint revision surgery. 34 patients had hip and 19 patients had knee joint revision arthroplasty, this includes single and multiple stage revision surgeries and excision arthroplasty. Nine (17%) patients had microbiologically proven joint infection. PV had sensitivity of 100%, specificity of 43% and negative predictive value of 100%. CRP had sensitivity of 89 %, specificity of 75% and negative predictive value of 97%. FS (presence of infection being more than 5 neutrophils/hpf) had sensitivity of 56% and specificity of 84%. We recommend PV and CRP to be used in the investigation of prosthetic joint infection. If both CRP and PV are normal the chance of infection is very low (negative predictive value of 100%). In our series an elevated PV and CRP represented a 50% chance of having a joint infection. The role of frozen section does not appear to be beneficial in the diagnosis of joint

The Masquelet or induced membrane technique (IMT) is a two-stage surgical procedure used for the treatment of segmental bone defects. In this technique, the defect is first filled with a polymethyl methacrylate (PMMA) spacer, which triggers the formation of a membrane that will encapsulate the defect. During the second surgery, the spacer is carefully removed and replaced by autologous bone graft while preserving the membrane. This membrane is vascularized, contains growth factors, and provides mechanical stability to the graft, all of which are assumed to prevent graft resorption and promote bone healing. The technique is gaining in popularity and several variations have been introduced in the clinical practice. For instance, orthopaedic surgeons now often include antibiotics in the spacer to treat or prevent infection. However, the consequences of this approach on the properties of the induce membrane are not fully understood. Accordingly, in a small animal model, this study aimed to determine the impact on the induced membrane of impregnating spacers with antibiotics frequently used in the IMT. We surgically created a five-mm segmental defect in the right femur of 25 adult male Sprague Dawley rats. The bone was stabilized with a plate and screws before filling the defect with a PMMA spacer. Animals were divided into five equal groups according to the type and dose of antibiotics impregnated in the spacer: A) no antibiotic (control), B) low-dose tobramycin (1.2 g/40 g of PMMA), C) low-dose vancomycin (1 g/40 g of PMMA), D) high-dose tobramycin (3.6 g/40 g of PMMA), E) high-dose vancomycin (3 g/40 g of PMMA). The animals were euthanized three weeks after surgery and the induced membranes were collected and divided for analysis. We assessed the expression of selected genes (Alpl, Ctgf, Runx2, Tgfb1, Vegfa) within the membrane by quantitative real-time PCR. Moreover, frozen sections of the specimens were used to quantify vascularity by immunohistochemistry (CD31 antigen), proliferative cells by immunofluorescence (Ki-67 antigen), and membrane thickness. Microscopic images of the entire tissue sections were taken and analyzed using FIJI software. Finally, we measured the concentration of vascular endothelial growth factor (VEGF) in the membranes by ELISA. No significant difference was found among the groups regarding the expression of genes related to osteogenesis (Alpl, Runx2), angiogenesis (Vegfa), or synthesis of extracellular matrix (Ctgf, Tgfb1) (n = four or five). Similarly, the density of proliferative cells and blood vessels within the membrane, as well as the membrane thickness, did not vary substantially between the control, low-dose, or high-dose antibiotic groups (n = four or five). The concentration of VEGF was also not significantly influenced by the treatment received (n = four or five). The addition of tobramycin or vancomycin to the spacer, at the defined low and high doses, does not significantly alter the bioactive characteristics of the membrane. These results suggest that orthopaedic surgeons could use antibiotic-impregnated spacers for the IMT without compromising the induced membrane and potentially bone healing

Aim. The aim of the study is to evaluate the specificity and sensibility of leukocyte esterase for the diagnosis of periposthetic joint infection (PJI). Method. Between October 2016 and April 2017 we enrolled 65 patients underwent to hip and knee revision arthroplasty due to uncertain joint infection. Synovial fluid was obtained from 64 joints that underwent revision arthroplasty. Each patient was evaluated in the preoperative time with CRP, ESR and leukoscan, in the intraoperative time with frozen section and leukocyte esterase strip and post-operative with sonication fluid culture, periprosthetic tissues cultures and histological examination. Results of all of these exams were compared to assess the specificity, the sensibility, the positive and negative predicting values of leukocyte esterase for the diagnosis of PJI. Results. The leukocyte esterase test with a threshold of +/++ had a sensitivity of 80.2%, a specificity of 82.8%, a positive predictive value of 63.8%, and negative predictive value of 92.1%. Using the threshold of ++ as a positive leukocyte esterase result, the specificity reached 97.8%, the positive predictive value 90.8%, and the negative predictive value 89.0%. Conclusions. These results demonstrate that leukocyte esterase is a quite accurate, effective marker of periprosthetic joint infection and that it is a valuable tool that can be used in conjunction with the the other tests for diagnosis of PJI

Infection is a devastating complication following total hip arthroplasty. For chronically infected total hip arthroplasty; we utilise a 2-stage articulating antibiotic hip spacer technique. Our success rate with the technique is 90% with patients clinically free of infection at an average of five years postoperatively. 80% of the patients had a positive identification of the infecting organism. The other 20% had a positive frozen section and clinically infected joints. All patients receive a minimum of 6 weeks of intravenous antibiotics. Postoperative hip scores average 90 points. Some patients elected to keep their antibiotic spacers for up to 2 years before the 2nd stage. Advantages of this technique include improved patient function, maintenance of bone stock and soft tissue tension, thus simplifying reimplantation

Aim. Quantitative assessment of alpha-Defensin offers a promising approach for diagnosing a periprosthetic joint infection (PJI) with sensitivities and specificities ranging from 97% to 100% and 95% to 100%, respectively. However, to the best of our knowledge and after due inquiry little information exists concerning qualitative measurements of alpha-Defensin. The aim of this study was to assess the diagnostic accuracy of the alpha-Defensin test, a lateral flow test for the qualitative detection of alpha-Defensin. Method. In this study, 50 patient with indicated revision surgery met the inclusion criteria due to septic or aseptic loosening. In addition to clinical standard diagnostics of PJI, the alpha-Defensin test. *. for the assessment of the qualitative alpha-Defensin in the synovial fluid was performed. The results were compared with the sensitivity and specificity of currently available clinical tests, specifically C-reactive protein (CRP), frozen section, definitive histology, bacteriology and sonication. Results. Based on the Musculoskeletal Infection Society's (MSIS) definition of PJI, 36 cases were categorized as aseptic and 13 as septic revisions. Due to the lack of an indicated control line (“C”), one alpha-Defensin test. *. was inconclusive. Qualitative alpha-Defensin had an area under the curve, sensitivity, specificity, and positive and negative likelihood ratios of 0.82, 69%, 94%, 12.46, and 0.33, respectively. Adjusted p-values using the method of Hochberg showed that the alpha-Defensin test. *. is significantly at least as good when diagnosing PJI as histology (p=0,0042) and bacteriology with at least one positive culture (p=0,0327). Conclusions. Qualitative alpha-Defensin tests could be an effective supplement in diagnosing PJI with a diagnostic accuracy comparable to histology and bacteriology (≥ 1 positive culture)

The practice at most centers in North America for the investigation and management of non-acute infection after hip replacement has been relatively standard for some time. Diagnosis has depended on a thorough history, physical examination, plain radiographs, straightforward laboratory inflammatory markers, joint aspiration for bacteriologic study, intraoperative frozen section in selected cases, and intraoperative synovial biopsies for confirmatory bacteriologic evaluation. The cornerstone of treatment on this continent has revolved around two-stage revision hip replacement, with increasing popularity for the use of interval articulating antibiotic loaded spacers, and increasing use of cementless fixation at the second stage. But this standard approach has been under increasing scrutiny in recent years, for good reason. The use of more precise “best evidence” paradigms on which to base the diagnosis have been developed. There is encouraging work on the application of more specific synovial and serum markers. The need to remove all implant material in all cases has been challenged. And there is evidence that the two stage approach is associated with greater morbidity, mortality and cost. The latter has led to a re-examination of the role and results of single-stage exchange revision, at least in selected cases, where the patient is immune competent, the soft tissue and bone anatomy is not badly deficient, the organism is known, and the antibiotic sensitivity is favorable. The most encouraging of the recent developments is the increasing consensus that multicenter collaborative study is required if we are to make genuine progress in the one-stage/two-stage debate. At least one multicenter prospective randomised study is scheduled to commence in 2015

There are unfortunately many reasons a TSA can be painful after implantation, but the most common reason is sepsis. Making the diagnosis can be a major challenge, but the biggest challenge is to think of that as a diagnosis! The most important steps are to first obtain plain radiographs one week after surgery so that you can compare subsequent radiographs to the immediate post-operative films. Progressive radiolucent lines in the glenoid or especially around the humerus are important hints. A loose humeral component is infected until proven otherwise. Next blood work to include CRP and ESR are critical. Other markers of infection have not been used on a widespread basis. If there is concern that there might be rotator cuff pathology and not sepsis, then we obtain an arthrogram CT scan at the time of aspiration. A cell count is helpful but often there is a dry tap. It is important to create a “p. acnes protocol” at your hospital to take cultures out 15 days. If still not sure and revision is necessary then we aspirate the joint at the time of surgery for cell count. The WBC cell count at the time of surgery keeps changing but over 3000 is considered diagnostic. Multiple specimens are sent for frozen section, culture and permanent section. The more WBC per high powered field the more likely there is an infection. Gram stains are worthless and we do not rely on them. We have no experience with implant sonification or use of IL-6

Diagnosis of chronic prosthetic joint infection (PJI) is often challenging. Painful prosthesis is frequently due to an infection but to diagnose it is somethimes difficult. All recent guidelines stress the central role of joint punction in diagnosis of PJI if the infection is not demonstrated. However which test on synovial fluid must be carried out is not so clearly defined. Total white blood cell count and differential leukocite count are usually considered useful in diagnosis but cut offs reported by different studies are quite different. Moreover this test needs a relatively large amount of fluid and blood contamination of it largely affects the result. What's more the synovial fluid WBC count may be unreliable in the setting of a metal-on-metal bearing or corrosion reaction. Routine cultures should be maintained between 5 and 14 days, their sensitivity appears low in chronic infection even if witholding antimicrobial therapy before the collection of the fluid can increase the likelihood of recovery an organism. Synovial leukocyte esterase can be performed as a rapid office or intraoperative point of care test using urinalysis strips. It is cheap and easy to perform, but the presence of blood in the sample can affect the result and it needs centrifugation. Recently a new test has been proposed to detect alfa-defensine in synovial fluid. It shows a high sensitivity and an exellent specificity. We performed 25 joint punctions on 25 patients with suspected PJI (enrollment is going on). Synovial fluid collected was tested for: leukocite esterase, WBC count and differential, colture in blood colture bottle for anerobe and aerobes (BacT/ALERT Biomerieux, inc) and detection of alfa-defensine level (Synovasure – Zimmer). In patients who underwent surgery at least 5 samples of periprotesic tissue were collected for microbiologic analysis and the removed implant was sonicated according with the methodic. Furthermore samples for frozen section were sent and a histologic examination was made according to the Moriewitz – Kerr classification. The MSIS criteria was utilized to classify the case as infected or not

The infected joint arthroplasty continues to be a very challenging problem. No test has 100% diagnostic accuracy for PPI and the treating surgeon must correlate the clinical and radiographic presentation with a combination of blood tests, synovial fluid analysis, microbiological and histopathological evaluation of periprosthetic tissue and intra-operative inspection to reach a definitive diagnosis. Diagnosis should begin with a high index of suspicion for new onset of pain or symptoms in well-functioning joints. Plain radiographs may identify osteolysis or early signs of implant failure and should be promptly investigated further for PPI. Peripheral blood ESR and CRP remain the most widely used next step for the diagnosis of PPI. Both these tests are widely available, inexpensive, and have a rapid turnaround time in laboratories. The results should be interpreted with caution due to their relative lack of specificity. The sensitivity and specificity values for CRP are approximately 88 and 74%, respectively; while that of ESR is slightly lower at 75 and 70%, respectively. The combined ESR and CRP tests are 96% sensitive for ruling out PPI but the specificity of this combination is as low as 56%. Advanced imaging modalities may be used as a part of the diagnostic algorithm. However, they require expert interpretation and are limited by availability and high costs. When available they have high sensitivity and specificity but their routine use is not recommended and indications have to be individualised in the light of clinical presentation. In the presence of high clinical suspicion, the clinician should plan synovial fluid analysis. This provides a synovial fluid white cell count with differential cell count, specimen for culture and possibility of analyzing other synovial fluid markers. It is important to note that failed metal-on-metal hip arthroplasties can give a falsely elevated synovial fluid cell count when using automated cell counters. This can be overcome by manually counting cell numbers. Synovial fluid should be directly into blood culture bottles, and antibiotics should be withheld at least 2 weeks prior to aspiration, whenever possible. Cultures also help establish the organism, virulence and sensitivities that help plan subsequent treatment algorithm. Periprosthetic tissue biopsy provides valuable information in microbiological diagnosis and workup of PPI. Routine use of gram staining is not recommended due to poor sensitivity. However, frozen section may have some role especially when performed by a skilled pathologist. Tissue culture remains the gold standard for diagnosis despite false-positive and false-negative results. Whenever possible multiple samples should be obtained to aid interpretation. A threshold of 2 to 3 positive specimens yielding indistinguishable microorganisms has been recommended to improve sensitivity. Acute inflammation, evidenced by neutrophilic infiltrate on fixed or frozen tissue, is suggestive of PPI and is defined as the presence of at least 5 neutrophils per high-powered field, in at least 5 separate microscopic fields. Sonication of removed prosthetic components is used to dislodge the biofilm and the associated bacteria from the surface of the implant. The fluid surrounding the implant can be used for culture or analysis. PCR testing: Synovial fluid aspirate, periprosthetic tissue or sonicate fluid may be subject to molecular diagnosis to amplify genetic material and improve microbiological diagnosis of PPI. This technique has shown increased sensitivity in patients who had received antibiotics within 14 days before implant removal. Results have to carefully interpreted with due consideration for possibility of false positive results

Periprosthetic joint infection (PPJI) following shoulder arthroplasty is uncommon, with an overall rate of 0.98%. However, the rates following revision arthroplasty and reverse arthroplasty are much higher. Given the rapid increase in the prevalence of shoulder arthroplasty and the increasing revision burden, the cost of PPJI to society will likely increase substantially. The most common organisms found in PPJI following shoulder arthroplasty are Staphylococcus aureus, coagulase-negative Staphylococcus, and Propionibacterium acnes (P. acnes). P. acnes is especially common in males. Traditional testing for PPJI includes aspiration, white blood cell count (WBC), erythrocyte sedimentation rate (ESR), and c-reactive protein (CRP). Aspiration often yields a dry tap and when fluid is obtained for culture, a positive result is helpful but a negative result does not rule out PPJI. Although WBC, ESR, and CRP are often positive with PPJI in the lower extremity, they are most often negative in shoulder PPJI. Although bone scans and WBC labeled scans are used, they are expensive and have low sensitivity and specificity. New testing and techniques have been reported in an attempt to improve sensitivity and specificity for PPJI. These techniques can be divided into tests on serum, synovial fluid, and tissue. Serum Interleukin-6 (IL-6) is highly specific (94%) for shoulder PPJI but has low sensitivity (14%). Synovial fluid can be tested for leukocyte esterase using a simple and cheap technique. In lower extremity PPJI it has shown to be helpful. It is not as helpful in shoulder PPJI with 30% sensitivity and 67% specificity. Alpha defensin has been reported to be more sensitive (63%) and as specific (95%) as traditional techniques but still lacks predictive value. Testing for specific cytokines (IL-2, IL-6, TNF- α) within synovial fluid is not widely used as yet but has shown promise with 80% sensitivity and 90% specificity. Obtaining tissue for culture and other testing is probably the most reliable way of confirming PPJI for the shoulder. Frozen sections taken at the time of revision can be helpful but is very pathologist dependent and institution specific. With a dedicated musculoskeletal pathologist, the finding of 10 or more WBCs per high powered field has been reported to be 72% sensitive and 100% specific for P. acnes and 63% sensitive and 100% specific for other organisms. Cultures from arthroscopic tissue biopsy have also been found to have high sensitivity (100%) and specificity (100%). Genetic testing of tissue biopsy specimens (PCR/NGS) has recently been reported and shows great promise. The significance of positive cultures and other tests, especially for P. acnes is unclear. There is a high rate of positive intra-operative cultures in primary cases of shoulder arthroplasty. In addition, intra-operative cultures taken at the time of revision, even in cases in which infection is not suspected, are frequently positive for P. acnes with weak correlation with rates of post-operative clinical infection. In conclusion, shoulder PPJI is a difficult problem to deal with. The definition of shoulder PPJI is currently unclear and further study is needed. There is no ideal test to confirm it. A reasonable approach is to aspirate for culture, and perform serum tests for WBC, ESR, and C-reactive protein. If any of these is positive in the setting of a painful arthroplasty, PPJI should be assumed until proven otherwise. Operative tissue cultures are probably the most reliable test but the clinical significance is not always obvious. Synovial fluid cytokine profiles and tissue PCR/NGS show promise for the future