Aim. Periprosthetic joint infection (PJI) is one of the most serious and frequent complications in prosthetic surgery. Despite significant improvements in the criteria for diagnosis of PJI, the diagnostic workflow remains complex and, sometimes, inconclusive. Host immune factors hold great potential as diagnostic biomarkers in bone and joint infections. We have recently reported that the synovial concentration of the humoral pattern recognition molecule long pentraxin 3 (PTX3) is a sensitive and specific marker of PJI in total hip and knee arthroplasty patients (THA and TKA) undergoing revision surgery [1]. However, the contribution to risk and diagnosis of PJI of the genetic variation in PTX3 and inflammatory genes that are known to affect its expression (IL-1b, IL-6, IL-10, and IL-17A) has not been addressed. Therefore, we assessed these relationships in a cohort of THA and TKA patients who underwent prosthesis revision by focusing on a panel of single nucleotide polymorphisms (SNPs) in the PTX3, IL-1β, IL-6, IL-10 and IL-17A genes. Method. A case-control retrospective study was conducted on an historic cohort of patients that received THA or TKA revision and were diagnosed with PJI (cases) or aseptic complications (controls) [1]. Samples of saliva were collected from 93 subjects and used for extraction of genomic DNA to perform genotyping of the PTX3, IL-1β, IL-6, IL-10 and IL-17A polymorphisms. Moreover, whenever available, samples of synovial fluid and plasma [1] were used to measure the concentration of the IL-1β, IL-10, and IL-6 proteins by immunoassay. Uni-and multivariate analyses were performed to evaluate the relationships between genetic, biochemical, and clinical variables. Results. The rs3024491 (IL-10) and rs2853550 (IL-1b) SNPs were found to be strongly associated with the risk of PJI. The synovial levels of PTX3, IL-1β, IL-10, and IL-6 were higher in cases than in controls, and a clear correlation emerged between the synovial concentration of PTX3 and IL-1b in cases only. Also, we identified a causal relationship between rs2853550, synovial concentration of IL-1b and that of PTX3 (that is induced by IL-1b). Conclusions. Our findings suggest that SNPs in the IL-10 and IL-1b genes could be used for early identification of THA and TKA patients with high risk of PJI. It is therefore conceivable that integrating genetic data into current diagnostic criteria would improve diagnosis of PJI

Aim. Unexpected negative-cultures (UNC) are a common diagnostic problem in periprosthetic joint infection (PJI) of the hip and knee when using culture-based methods. A novel molecular approach (MC)1 based on the identification of the vast majority of bacterial species in a single assay using species-specific bacterial interspacing region length polymorphisms and phylum-specific 16S rDNA sequence polymorphisms has demonstrated clinical utility in PJI diagnostics (1). In addition, MC provides an estimate of the leukocyte concentration in the specimen analysed. The aim of this retrospective, blinded study was to evaluate the performance of MC in identifying the microbiological content and determining the leukocyte count in synovial fluid (SF) collected from hip and knee revision arthroplasty cases with UNC. It was also assessed whether antibiotic treatment would have been changed if the result from MC had been known. Method. A total of 89 SF samples from 70 patients (43 female; 27 male) who underwent revision arthroplasty (14 hip; 75 knee) were included. Using European and Bone Joint Infection Society (EBJIS) criteria, 82 cases were classified as infected (77 UNC and 5 septic culture-positive controls), five as non-infected (aseptic culture-negative controls), and two as likely infected, but infected by clinical observation. MC was performed and evaluated together with SF parameters. Antibiotic treatment, clinical outcome, patient demographics and surgical details were analysed. Results. Overall, 29.1% (23/79) of UNC had a positive yield by MC, of which 2/23 (8.7%) had two microorganisms detected simultaneously. Of the 25 microorganisms identified by MC, 12/25 (48%) were clinically relevant after re-evaluation of the patients’ microbiological history. The microorganisms detected were 5/25 (20%) Streptococcus pneumoniae/mitis, 4/25 (16%) Staphylococcus epidermidis, 3/25 (12%) Cutibacterium acnes, 3/25 (12%) Streptococcus agalactiae, 2/25 (8%) Streptococcus bovis, 2/25 (8%) Staphylococcus aureus, and 2/25 (8%) Haemophilus parainfluenzae. The prevalence of Enterococcus faecalis, Bacteroides fragillis, Staphylococcus lugdunensis, Corynebacterium striatum among all MC results was 1/25 (4%) each species. In total, 13/23 (56%) cases were associated with patients receiving antibiotic therapy at the time of SF collection. The yield for leukocyte counts provided the molecular technique was consistently much higher in the UNC and clearly septic groups than in the clearly aseptic group. Overall, 20/61 (32.8%) patients with UNC could have been managed differently and more accurately after MC assessment. Conclusions. MC shows clinical value in the diagnosis and management of PJI with UNC. The included leukocyte count shows promising results. Acknowledgments. This work was partially funded by Inbiome

Aim. Previous studies had indicated that interleukin-1 beta (IL-1β) gene single nucleotide polymorphisms (SNPs) associate with different inflammatory diseases. However, potential links between these polymorphisms and susceptibility to extremity chronic osteomyelitis (COM) in Chinese population remain unclear. This study aimed to investigate relationships between IL-1β gene polymorphisms (rs16944, rs1143627, rs1143634 and rs2853550) and the risk of developing extremity COM in Chinese population. Method. Altogether 233 extremity COM patients and 200 healthy controls were genotyped for the four tag SNPs of the IL-1β gene using the SNapShot genotyping method. Comparisons were performed regarding genotype distribution, mutant allele frequency and four genetic models (dominant, recessive, homozygous and heterozygous models) of the 4 SNPs between the two groups. Results. Significant associations were identified between rs16944 polymorphism and the risk of developing COM by dominant model (P = 0.026, OR = 1.698, 95% CI 1.065–2.707) and heterozygous model (P = 0.030, OR = 1.733, 95% CI 1.055 – 2.847). Although no statistical differences were found of rs1143627 polymorphism between the two groups, there existed a trend that rs1143627 may be linked to an elevated risk of developing COM by outcomes of dominant (P = 0.061), homozygous (P = 0.080) and heterozygous (P = 0.095) models. However, no statistical correlations were found between rs1143634 and rs2853550 polymorphisms and susceptibility to COM in Chinese population. Conclusions. To our knowledge, we reported for the first time that IL-1β gene rs16944 polymorphism may contribute to the increased susceptibility to extremity COM in Chinese population, with genotype of AG as a risk factor

Aim. Previous studies have indicated that TNF-α and lymphotoxin-α (LTA) gene polymorphisms associate with the development of several different inflammatory diseases. However, potential associations of such gene polymorphisms with the susceptibility to extremity chronic osteomyelitis remain unknown. This study aimed to investigate potential links between TNF-α gene polymorphisms (rs1800629, rs361525, rs1799964, rs1800630, rs1799724 and rs1800750) and LTA gene polymorphism (rs909253) and the risk of developing extremity chronic osteomyelitis in Chinese population. Method. A total of 233 patients with extremity chronic osteomyelitis and 200 healthy controls were genotyped for the above 7 polymorphisms of TNF-α and LTA genes using the genotyping method*. Results. Significant difference was found regarding the genotype distribution of rs909253 between patients and healthy controls (P = 0.002). The mutant allele C frequency of rs909253 in patient group was significantly higher than that in control group (P = 0.001). Significant associations were identified between rs909253 and the risk of developing chronic osteomyelitis by dominant model (P = 0.040), recessive model (P = 0.002) and homozygous model (P = 0.001). Additionally, the mutant allele T frequency in rs1799964 in patient group was significantly higher than that in control group (P = 0.035). Significant link was found between rs1799964 and susceptibility to chronic osteomyelitis by recessive model (P = 0.048). However, no significant outcomes were identified regarding other TNF-α gene polymorphisms between the two groups. Conclusions. The present study demonstrated that rs909253 and rs1799964 polymorphisms may associate with the risk of developing chronic osteomyelitis in Chinese population. *SNaPshot

Aim. Cyclooxygenase-2 (COX-2) enzyme is one of the major mediators during inflammation reactions, and COX-2 gene polymorphisms of rs20417 and rs689466 have been reported to be associated with several inflammatory diseases. However, potential links between the two polymorphisms and risk of developing post-traumatic osteomyelitis remain unclear. The present study aimed to investigate associations between the rs20417 and rs689466 polymorphisms and susceptibility to post-traumatic osteomyelitis in Chinese population. Methods. A total of 189 patients with definite diagnosis of post-traumatic osteomyelitis and 220 healthy controls were genotyped for rs20417 and rs689466 using the genotyping method*. Chi-square test was used to compare differences of genotype distributions as well as outcomes of five different genetic models between the two groups. Results. Significant association was found between rs689466 and post-traumatic osteomyelitis by recessive model (GG vs. AA + AG) (OR = 1.74, 95% CI: 1.098–2.755, P =0.018). Although no statistical differences were identified of rs689466 between the two groups by allele model (P = .098) or homozygous model (P = 0.084), outcomes revealed a tendency that allele G may be a risk factor and people of GG genotype may be in a higher risk to develop post-traumatic osteomyelitis in Chinese population. However, no significant link was found between rs20417 and susceptibility to post-traumatic osteomyelitis in this Chinese cohort. Conclusions. To our knowledge, we reported for the first time that COX-2 gene polymorphism rs689466 may contribute to the increased susceptibility to post-traumatic osteomyelitis in Chinese population. *SNaPshot®

Toll-like receptors (TLRs) are crucial components of the immune system that recognize microbial infection and trigger anti-microbial host defense responses. Gram positive bacteria are causative factors of bone infections, as they alter the balance of coordinated activities during bone remodeling, stimulating osteoclastogenesis. The aim of the study was to investigate whether genetic variation in TLR2 and TLR4 genes predisposes to bone infections’ susceptibility. One hundred and twenty patients with bone infections (osteomyelitis) and 200 healthy controls were genotyped for two single nucleotide polymorphisms (SNPs), R753Q [A/G] in TLR2 gene and T399I [C/T] in TLR4 gene. DNA was extracted from whole blood and the above SNPs were typed with PCR-RFLP (Polymerase Chain Reaction- Restriction Fragment Length Polymorphism) method for genotype identification. All patients were infected by Gram-positive bacteria, predominantly Staphylococcus aureus. Statistical analysis was carried out using the chi-square test. We observed a significantly increased frequency in patients carrying the GA genotype of TLR2 R753Q polymorphism compared to controls (p<0.05). We also found that the A allele was more common in patients than in controls. All individuals carrying the A allele were heterozygous for this variant, while homozygous mutant individuals were not detected in the patients and the control group. In contrast, we found that the TLR4 T399I [C/T] SNP was similarly distributed among the two groups (patients and controls). The mechanism through which TLR2 mediates its effect in bone infections is under investigation. A significant difference was observed in the genotype frequency of TLR2 R753Q [A/G] polymorphism in patients, suggesting that genetic variability in TLR2 gene may be associated with susceptibility to osteomyelitis in response to bacterial invasion in the bone

Introduction. This study was to investigate the association of developmental dysplasia of the hip (DDH) and primary protrusion acetabuli (PPA) with Vitamin D receptor polymorphisms TaqI and FokI and oestrogen receptor polymorphisms Pvu II and XbaI. Methods. 45 patients with DDH and 20 patients with PPA were included in the study. Healthy controls (n=101) aged 18-60 years were recruited from the same geographical area. The control subjects had a normal acetabular morphology based on a recent pelvic radiograph performed for an unrelated cause. DNA was obtained from all the subjects from peripheral blood. Genotype frequencies were compared in the three groups. The relationship between the genotype and morphology of the hip joint, severity of the disease, age at onset of disease and gender were examined. Results. The oestrogen receptor XbaI wild-type genotype (XX, compared with Xx and xx combined) was more common in the DDH group (55.8%) than controls (37.9%), though this just failed to achieve statistical significance (p=0.053, odds ratio=2.1, 95% CI=0.9-4.6). In the DDH group, homozygosity for the mutant TaqI Vitamin D receptor t allele was associated with higher acetabular index (Mann-Whitney U-test, p= 0.03). Pvu II pp oestrogen receptor genotype was associated with low centre edge angle (p=0.07). Conclusion. This study suggests a possible correlation between gene polymorphism in the oestrogen and vitamin D receptors and susceptibility to, and severity of DDH. The TaqI vitamin D receptor polymorphisms may be associated with abnormal acetabular morphology leading to DDH while the XbaI oestrogen receptor XX genotype may be associated with increased risk of developing DDH. No such correlations were found in the group with PPA

While total joint replacement (TJR) is considered as an effective intervention to relieve pain and restore joint function for end-stage osteoarthritis (OA) patients, a significant proportion of the patients are dissatisfied with their surgery outcomes. The aim of this study was to identify genetic factors that can predict patients who do or do not benefit from these surgical procedures by a genome-wide association study (GWAS). Study participants were derived from the Newfoundland Osteoarthritis Study (NFOAS) which consisted of 1086 TJR patients. Non-responders to TJR was defined as patients who did not reach the minimum clinically important difference (MCID) based on the self administered Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) in terms of pain reduction or function improvment. DNA was extracted from the blood samples of the study participants and genotyped by Illumina GWAS genotyping platform. Over two million single nucleotide polymorphisms (SNPs) across the genome were genotyped and tested for assocition with non-responders. 39 non-responders and 44 age, sex, and BMI matched responders were included in this study. Four chromosome regions on chromosomes 5, 7, 8, and 12 were suggested to be associated with non-responders with p < 1 0–5. The most promising one was on chromosome 5 with the lead SNP rs17118094 (p=1.7×10–6) which can classify 72% of non-responders accurately. The discriminatory power of this SNP alone is very promising as indicated by an area under the curve (AUC) of 0.72 with 95% confidence interval of 0.63 to 0.81, which is much better than any previously studied predictors mentioned above. All the patients who carry two copies of the G allele (minor allele) of rs17118094 were non-responders and 75% of those who carry one copy of the G allele were non-responders. The discriminatory ability of the lead SNPs on chromosomes 7 and 12 were comparable to the one on chromosome 5 with an AUC of 0.74, and 88% of patients who carry two copies of the A allele of rs10244798 on chromosome 7 were non-responders. Similarly, 88% of patients who carry two copies of the C allele of rs10773476 on chromosome 12 were non-responders. While the discriminatory ability of rs9643244 on chromosome 8 was poor with an AUC of 0.26, its strong association with non-responders warrants a further investigation in the region. The study identified four genomic regions harboring genetic factors for non-responders to TJR. The lead SNPs in those regions have great discriminatory ability to predict non-responders and could be used to create a genetic prediction model for clinical unitilty and application

Introduction. Previous hemodynamics studies in shoulder arthroplasty only evaluated Western population and mainly focused on risk factors of transfusion. However, Asians are relatively small, and have higher bleeding risk due to prothrombin-clotting-factor polymorphisms. Therefore, it is not appropriate to apply the results of previously studies directly to Asians. Authors compare different hemodynamics depending on the types of shoulder arthroplasties, and evaluate predictors for transfusion in Asian population. Methods. Total 212 shoulder arthroplasties (26 fracture hemiarthroplasty (fHA), 49 anatomical total shoulder arthroplasty (aTSA), 132 reverse total shoulder arthroplasty (rTSA), and 5 revision surgery) from August 2004 to January 2016 were retrospectively reviewed. Demographics, surgical factors and perioperative hemodynamic factors were compared for each type of arthroplasty. Multivariate regression analysis was conducted to determine predictors for transfusion. Results. Preoperative Hb and Hct level were lower in fHA group (11.9 ± 1.8g/dL, 35.1 ± 5.4%; p < 0.001, 0.001). Total drain output was higher in rTSA and revision (349.1 ± 191.7mL, 408.6 ± 125.8mL; p<0.001, <0.001), however, there was no significant difference of estimated blood loss (p=0.269). There was significant, but very weak correlation between drain output with Hb decrease in postoperative one day (r = 0.192; p = 0.007). However, there were not significant correlation between drain output with Hb decrease after postoperative 2 and 3 days (r = 0.185, 0.001; p = 0.241, 0.997). The overall transfusion rate was 11.3% (24/212); fHA 30.8% (8/26), aTSA 10.2% (5/49), rTSA 7.6% (10/132), revision 20% (1/5). In multivariate regression analysis, lower Hb level of preoperative period and postoperative 1 day were predictors for transfusion (OR = 0.481, 0.499; p = 0.002, 0.017) and cutoff-value were 12.15g/dL and 10.0g/dL, respectively (OR = 7.404, 5.499; p = <0.001, 0.001; Sensitivity=80%, 70%; Specificity=80%, 84%). Conclusion. In Asian, overall transfusion rate after shoulder arthroplasties was 11.3%, varied by type of arthroplasty. Lower Hb level of preoperative period (<12.15g/dL) and postoperative 1 day (<10.0g/dL) were predictors for transfusion. Surgeons should consider different hemodynamics depending on different types of shoulder arthroplasty, and close monitoring of perioperative Hb level is essential to decrease hemorrhage-related complications, because drain output could not represent perioperative hemorrhage

Propionibacterium acnes infection of the shoulder after arthroplasty is a common complication. Current detection methodologies for P. acnes involve prolonged anaerobic cultures that can take up to three weeks before findings can be reported. Our aim was to develop a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) approach that is both sensitive and specific to P. acnes that would enable a 24-hour turnaround between biopsy and results. Comparisons between the 16S ribosomal sequences of P. acnes and closely related bacteria identified two unique regions in P.acnes to which PCR primers were designed. Additionally, two unique restriction enzyme cut sites for HaeIII were identified within this amplicon. To test the PCR method, arthroscopic surgical biopsies were mechanically homogenised and boiled for 20 minutes to lyse the cellular membranes. PCR was performed using standard conditions followed by a one hour HaeIII enzymatic digest of the PCR product. Resultant fragments were visualised on polyacrylamide gels stained with ethidium bromide. All experiments included no-template controls to rule out reagent contamination and independently confirmed P. acnes DNA as a positive control. Serial dilutions of P. acnes cultures in Robertson's cooked-meat broth and spectrophotometric analysis of cellular concentration were used to assess the sensitivity of the PCR reaction. A unique 564 base-pair PCR amplicon was derived from different strains of P. acnes. This amplicon was confirmed as P. acnes DNA by gel excision and DNA sequencing. HaeIII digests of the amplicon yielded 3 restriction fragments at the sizes predicted by in silico analyses. Sensitivity testing confirmed that as few as 10 P. acnes cells in a 50µl reaction volume could be detected using this assay. P. acnes was also detected in surgical biopsy samples. P. acnes infections following shoulder arthroplasty are a serious complication placing a burden on the healthcare system and the patient due to the lengthy surgical revision process that follows. The infections are also difficult to diagnose. This unique assay combines the sensitivity of PCR with the specificity of RFLP mapping to specifically identify P. acnes in surgical isolates. We anticipate that this assay will allow us to determine if a biopsy is P. acnes positive within 24-hours of sampling, allowing for more aggressive antibiotic therapy and monitoring to avoid implant failure and revision surgery. Additionally, this PCR-RFLP method may decrease the false positive rate of extended length cultures due to P. acnes contamination

We studied twelve parameters (physical appearance, mucin clot, fibrin clot, white cell count, differential count, red blood cell count, gram stain for bacteria, crystal microscopy, aerobic bacterial culture, anaerobic bacterial culture and ratio between synovial sugar and blood sugar) in over 300 samples of synovial fluid from patients with a variety of suspected pathologies (e.g. infection, inflammatory disease, infection adjacent to a joint, aseptic loosening of a prosthesis). The diagnosis of infection was further established using clinical signs, radiological features, full blood count, C-reactive protein and iron profile. Many of the patients came to surgery. This of course created further opportunity to establish or rule out the diagnosis of infection with greater certainty. Nine of the features of synovial fluid were analysed statistically, including turbidity, diminished viscosity, mucin clot, fibrin clot, total white cell count, polymorphs greater than 60%, bacteria observed on direct microscopy, bacteria yielded by culture and concentration of synovial sugar less than 40% of the simultaneous blood sugar. The positive or negative features of infection were determined to be true or false in the light of the cumulative overall features of infection. The data so obtained was analysed to establish sensitivity, specificity, positive predictive value, negative predictive value and accuracy. The mass of data so obtained cannot be meaningfully expressed in such a brief abstract. Important examples are when culturing synovial fluid there were 44% false negatives or no growth and 56% true positives. Looking at the ratio between synovial sugar and blood sugar we found that taking 40% as the critical value, this was 62% sensitive, the specificity was 89%, the accuracy was 73%, the positive predictive value was 89%, the negative predictive value was 62.4%. However we went further and separated those who were definitely infected or probably infected i.e. Groups 4 & 5 from those who were probably or definitely NOT infected according to the sum of clinical laboratory and radiological parameters. When thus separated the predictive value of a positive result was 100% in Group 4 & 5 and 0% in Group 1 & 2. The predictive value of a negative result in Group 1 & 2 was 98.7% accurate and 22.4% in Group 4 & 5

INTRODUCTION. Since the recall of some metal on metal (MoM) THR bearings, surgeons have seen patients with pain, elevated Co and Cr levels and adverse local tissue reactions (ALTR). While many variables may contribute to THR MoM failures, many times these variables are not present in patients who present with symptoms. We investigate the possible genetic predilection of a group of patients who were revised after MoM THR surgery for pain, high Co/Cr levels and ALTR. METHODS. IRB approval was obtained prior to our study. We have analyzed 19 control (asymptomatic MoM THR patients > 6 years after surgery) and 19 disease (revised MoM THR for high metal ions and ALTR). The 38 sample intensity files were subject to sample Quality Control (QC) using Contrast QC (< 0.4) with an Affymetrix Genotyping Console. The resulting 38 sample files with genotype calls were loaded and further analyzed using the Association Workflow in Partek Genomics Suite 6.6 (Partek, Missouri). Hardy-Weinberg equilibrium test was performed on the single nucleotide polymorphism (SNP) level. The difference between the observed and expected frequencies of each allele at each locus were tested by Fisher's exact test and χ2 test. To get the working SNP list, two filters were used: (1) a SNP no-call rate should be less than 5%, and (2) minor allele frequency of a SNP should be greater than 5%. After filtering, association analysis of the SNPS with disease was done using Chi. 2. Test. In this study, χ. 2. statistic was used to assess the difference in allele frequencies between the control and disease samples. The value of χ2 statistic, degrees of freedom, and the associated p-value for each SNP were calculated. Dot Plot was used to visualize the genotypes of all samples. To measure the non-random association of alleles at different loci, Linkage Disequilibrium analysis was performed using the neighborhood size of 20 and statistic r. 2. The resulting correlations show the value of r. 2. for SNPs. The r. 2. = 1 means that two SNPs are tightly associated. RESULTS. We found that several SNPs are linked to the revision disease group that showed evidence of metal sensitivity. Among them, a strong association in the disease group was found in a SNP called MS1. In the disease group 17/19 patients were either heterozygous or recessive homozygous for MS1, with 17/19 asymptomatic patients were of the homozygous dominant MS1 isoform. Based on the Linkage Disequilibrum analysis results, several other SNPs were also fund to be strongly correlated with the disease group (Fig 1). The controls had an average Co level of 2.4 and Cr level of 1.3 while the disease group 18 and 10.4 respectively. CONCLUSIONS. This study found a strong genetic relationship in a gene we designate as MS1 where the homozygous recessive and heterozygous isoform genotypes were found in the disease group of revised MoM THRs. A strong correlation of several SNPs was also found. This may be a good predictor of failures and an avenue for personalized choice of implants in the future. For figures/tables, please contact authors directly.

The Osteoprotegerin/RANK/RANKL system has been implicated in the biological cascade of events initiated by particulate wear debris and bacterial infection resulting in periprosthetic bone loss around loosened total hip arthroplasties (THA). Individual responses to such stimuli may be dictated by genetic variation and we have studied the effect of single nucleotide polymorphisms (SNPs) within these genes. We performed a case control study of the Osteoprotegerin, RANK and RANKL genes for possible association with deep sepsis or aseptic loosening. All patients included in the study were Caucasian and had had a cemented Charnley THA and polyethylene acetabular cup. Cases consisted of 91 patients with early aseptic loosening and 71 patients with microbiological evidence at surgery of deep infection. Controls consisted of 150 THAs that were clinically asymptomatic for over 10 years and demonstrated no radiographic features of aseptic loosening. DNA samples from all individuals were genotyped using Taqman allelic discrimination. The A allele (p<0.001) and homozygous genotype A/A (p<0.001) for the OPG-163 SNP were highly associated with aseptic failure. Additionally, the RANK-575 (C/T SNP) T allele (p=0.004) and T/T genotype (p=0.008) frequencies were associated with aseptic failure. No statistically significant relationship was found between aseptic loosening and the OPG- 245 or OPG-1181 SNPs. When the septic group was compared to controls, the frequency of the A allele (p<0.001) and homozygous genotype A/A (p<0.001) for the OPG-163 SNP were statistically significant. No statistically significant relationship was found between septic failure and the OPG- 245, OPG-1181 or RANK-575 SNPs. Aseptic loosening and possibly deep infection of THA may be under genetic influence to candidate susceptibility genes. SNP markers may serve as predictors of implant survival and aid pharmacogenomic prevention of THA failure

Introduction:. Wear particles cause aseptic loosening by stimulating macrophages to produce inflammatory cytokines. Recent studies indicate that Toll-like receptor 2 (TLR2) and TLR4 mediate macrophage responses to the wear particles [1–3]. TLR2 and TLR4 uniquely activate MyD88-dependent signaling via an additional adapter protein known as TIRAP/Mal [4]. Del Vescovo et al reported that three single nucleotide polymorphisms (SNPs) within the TIRAP/Mal gene associate with aseptic loosening in THA patients [5]. The goal of the current study was therefore to determine whether TIRAP/Mal mediates responses to orthopaedic wear particles. Methods:. Immortalized wild type (WT) and TIRAP/Mal knockout (KO) murine macrophages (Mfs) were incubated in the presence or absence of titanium (Ti) particles (1 × 10. 8. particles/cm. 2. [2]. Three types of particles were used as described previously [1,2]: Ti particles with adherent bacterial debris (38.3 Endotoxin Units/10. 9. particles), endotoxin-free Ti particles (<0.1 EU/10. 9. particles), and Ti particles with adherent lipopolysacharide (LPS, 32.8 EU/10. 9. particles). TNFa, IL-1b, and IL-6 mRNAs were measured by real-time PCR and the secreted cytokines were measured by ELISA. Particle-induced osteolysis in calvaria of TIRAP/Mal KO and WT mice was measured 7 days after particle implantation [1,2]. In vitro results are presented as mean ± SEM of 3–4 replicate experiments analyzed by two-way ANOVA with Bonferroni post-hoc corrections. In vivo results are presented as means of individual parietal bones ± SEM (n = 22) and analyzed by one-way ANOVA on ranks with Student Neuman-Keuls post-hoc corrections. * denotes p < 0.5, ** denotes p < 0.01, *** denotes p < 0. Results:. Ti particles with adherent bacterial debris induced substantial osteolysis and expression of TNFa, IL-1b, and IL-6 at both the mRNA and protein levels and all of those responses were significantly inhibited by TIRAP/Mal KO (Fig 1 & Fig 2). Endotoxin-free Ti particles had a small effect on osteolysis and cytokine mRNA expression that was not dependent on TIRAP/Mal (Fig 1 & data not shown). Adherence of highly purified LPS to the endotoxin-free particles reconstituted the stimulation of osteolysis and cytokine expression as well as the dependence on TIRAP/MAL (Fig 1 & data not shown). Specificity of the effects of TIRAP/Mal KO was demonstrated since responses induced by recombinant murine IL-1b were unaffected (data not shown). Discussion:. Our results are the first demonstration that TIRAP/Mal mediates effects of orthopaedic wear particles. TIRAP/Mal KO inhibited expression of TNFa by ∼50% and almost completely inhibited particle-induced osteolysis, as well as expression of IL-1b and IL-6. Our results, coupled with the genetic association of SNPs in human TIRAP/Mal with aseptic loosening [5], lead to two conclusions. First, activation of TIRAP/Mal likely contributes to aseptic loosening in patients. Second, pathogen-associated molecular patterns (PAMPs) also likely contribute to aseptic loosening since the results with endotoxin-free Ti particles demonstrate that adherent PAMPs are required for activation of TIRAP/Mal

Aims

The STRYDE nail is an evolution of the PRECICE Intramedullary Limb Lengthening System, with unique features regarding its composition. It is designed for load bearing throughout treatment in order to improve patient experience and outcomes and allow for simultaneous bilateral lower limb lengthening. The literature published to date is limited regarding outcomes and potential problems. We report on our early experience and raise awareness for the potential of adverse effects from this device.

Non-tuberculous mycobacterial (NTM) infection of the musculoskeletal tissue is a rare disease. An early and accurate diagnosis is often difficult because of the indolent clinical course and difficulty of isolating pathogens. Our goal was to determine the clinical features of musculoskeletal NTM infection and to present the treatment outcomes. A total of 29 patients (nine females, 20 males between 34 and 85 years old, mean age 61.7 years; 34 to 85) with NTM infection of the musculoskeletal system between 1998 to 2011 were identified and their treatment retrospectively analysed. Microbiological studies demonstrated NTM in 29 patients: the isolates were Mycobacterium intracellulare in six patients, M. fortuitum in three, M. abscessus in two and M. marinum in one. In the remaining patients we failed to identify the species. The involved sites were the hand/wrist in nine patients the knee in five patients, spine in four patients, foot in two patients, elbow in two patients, shoulder in one, ankle in two patients, leg in three patients and multiple in one patient. The mean interval between the appearance of symptoms and diagnosis was 20.8 months (1.5 to 180). All patients underwent surgical treatment and antimicrobial medication according to our protocol for chronic musculoskeletal infection: 20 patients had NTM-specific medication and nine had conventional antimicrobial therapy. At the final follow-up 22 patients were cured, three failed to respond to treatment and four were lost to follow-up. Identifying these diseases due the initial non-specific presentation can be difficult. Treatment consists of surgical intervention and adequate antimicrobial therapy, which can result in satisfactory outcomes.

Cite this article: Bone Joint J 2014;96-B:1561–5.

The most frequent cause of failure after total hip replacement in all reported arthroplasty registries is peri-prosthetic osteolysis. Osteolysis is an active biological process initiated in response to wear debris. The eventual response to this process is the activation of macrophages and loss of bone.

Activation of macrophages initiates a complex biological cascade resulting in the final common pathway of an increase in osteolytic activity. The biological initiators, mechanisms for and regulation of this process are beginning to be understood. This article explores current concepts in the causes of, and underlying biological mechanism resulting in peri-prosthetic osteolysis, reviewing the current basic science and clinical literature surrounding the topic.