Periprosthetic Membrane Research Articles

Accuracy of Serological Markers, Synovial Fluid, Microbiological Culture, and Histopathological Examination for Diagnosing Periprosthetic Knee Infection.

Objective This study assessed the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of serological tests, synovial fluid markers, microbiological tissue culture, and histopathological examination of the periprosthetic membrane in diagnosing periprosthetic knee infection. Methods This study is prospective, and it includes patients undergoing total knee arthroplasty revision surgery from November 2019 to December 2021. The analysis consisted of serological tests (erythrocyte sedimentation rate [ESR], C-reactive protein [CRP], and D-dimer), synovial fluid markers (leukocyte and polymorphonuclear cell counts), periprosthetic tissue culture, and histopathological examination of the periprosthetic membrane of all patients. Results Sixty-two patients had periprosthetic joint infection (PJI) according to the 2018 International Consensus Meeting criteria (infection group), while 22 subjects had no infection. ESR sensitivity and specificity were 83.6% and 45.4%, respectively. CRP sensitivity and specificity were 64.5% and 100%, whereas D-dimer sensitivity and specificity were 78.9% and 25%, respectively. Leukocyte count sensitivity and specificity were 75.6% and 100%, polymorphonuclear cell count sensitivity and specificity were 33% and 100%, respectively. Periprosthetic tissue culture sensitivity and specificity culture were, respectively, 77.4% and 100%. Histopathological examination sensitivity and specificity were 43.7% and 100%, respectively. Conclusions In our study, the total blood cell count in synovial fluid and microbiological cultures of periprosthetic tissues were the most accurate tests for PJI diagnosis. In contrast, polymorphonuclear cell percentage was the least accurate test for PJI diagnosis.

Roles of inflammatory cell infiltrate in periprosthetic osteolysis.

Classically, particle-induced periprosthetic osteolysis at the implant-bone interface has explained the aseptic loosening of joint replacement. This response is preceded by triggering both the innate and acquired immune response with subsequent activation of osteoclasts, the bone-resorbing cells. Although particle-induced periprosthetic osteolysis has been considered a foreign body chronic inflammation mediated by myelomonocytic-derived cells, current reports describe wide heterogeneous inflammatory cells infiltrating the periprosthetic tissues. This review aims to discuss the role of those non-myelomonocytic cells in periprosthetic tissues exposed to wear particles by showing original data. Specifically, we discuss the role of T cells (CD3+, CD4+, and CD8+) and B cells (CD20+) coexisting with CD68+/TRAP- multinucleated giant cells associated with both polyethylene and metallic particles infiltrating retrieved periprosthetic membranes. This review contributes valuable insight to support the complex cell and molecular mechanisms behind the aseptic loosening theories of orthopedic implants.

When prosthetic joint infection may lead to premalignant colorectal lesion detection

Osteoarthritis (OA) is a common musculoskeletal disorder that affects millions of people worldwide, and total knee arthroplasty (TKA) is a common treatment for advanced OA. However, joint replacement surgeries may lead to complications such as deep vein thrombosis (DVT), surgical site infection, and prosthetic joint infection (PJI). Our objective is to discuss a Streptococcus bovis group (SBG) PJI case after TKA and its possible consequences in clinical practice. We describe a 67-year-old female who underwent TKA and developed initial complications including wound hematoma, symptomatic anemia, and acute DVT. Further investigations because of knee pain persistence and inflammatory signs post TKA revealed the presence of Staphylococcus aureus in the synovial and scar tissues from the knee and SBG in the periprosthetic membrane from the revision surgery, as well as mitral infective endocarditis and a large stenosing villous polypoid lesion in the ascending colon. SBG osteoarticular infection post TKA is a rare condition, but it may be associated with severe diseases. This article highlights the link between SBG and colorectal cancer (CRC) and emphasizes the importance of an echocardiogram and a colonoscopy for early detection and prompt treatment of infective endocarditis and CRC. This diagnostic workup should be performed even in the absence of symptoms, for early diagnosis and increased chance of cure. A multidisciplinary approach, involving orthopedists, infectious disease specialists, cardiologists, and gastroenterologists, among others, is advised to effectively manage SBG infections and improve patient outcomes.

Particle disease and its effects on periarticular tissue

Particle disease is the condition caused by wear debris on surrounding tissues and influences the well-being of arthroplasty patients. This condition is multifactorial due to the type of bearing couple, head size and implant position. Subsequent periprosthetic osteolysis and soft tissue reactions, can lead to revision THA surgery. The periprosthetic synovial membrane (synovial-like interface membrane, SLIM) is used in diagnostics when the cause of implant failure is uncertain. Detailed analysis of synovial fluid and bone marrow could improve the diagnostic procedure and strengthen the cases for revision surgery and the underlying biology. Alarge number of research approaches on this topic have evolved and continue to be utilized in the clinic.

Vitamin E-added Highly Cross-Linked Polyethylene Decreases the Risk of Osteolysis in an In Vivo Arthroplasty Model.

Introduction Aseptic loosening is one of the most important complications of arthroplasty surgery. It is known that immune response against particles plays role in the pathogenesis of aseptic loosening. Polyethylene (PE) has an important place in these particles. There are limited in vivo studies examining aseptic loosening caused by PE residues. Objective The aim of the present study is to evaluate the aseptic loosening created by highly cross-linked PE (HXLPE) and vitamin E-added PE particlesin an in vivo knee prosthesis model. Materials and methods Thirty-nine male Sprague-Dawley rats, which were randomized into three groups, were included in the study. After surgical exposure of knee joints of rats, femoral intramedullary canals were drilled and instilled with isolated saline solution and saline solution that contained standard PE or vitamin E-added PE particles according to their groups. Afterwards, a titanium implant was placed on the femoral articular surface of each animal. Rats received intraarticular injections weekly of the same solution, which was initially instilled into their femoral canal. The rats were sacrificed at the end of the third week and then underwentradiological and histopathological evaluations. Result In histopathological evaluation, periprosthetic membrane formation, inflammatory cell change, and cellular damage of cartilage and bone tissue around the implant were assessed. There was a statistically lesser amount of cellular damage and periprosthetic membrane formation in the vitamin-E/HXLPE group compared to the HXLPE group (p=0.04, p=0.001). No significant difference was found between the PE groups with respect to inflammatory cells (p=0.715). Conclusions HXLPE caused more significant osteolysis compared to VE-HXLPE. Antioxidants in PE could provide a reduction in osteolysis and aseptic loosening.

Three to six tissue specimens for histopathological analysis are most accurate for diagnosing periprosthetic joint infection.

The aim of this study was to evaluate the optimal deep tissue specimen sample number for histopathological analysis in the diagnosis of periprosthetic joint infection (PJI). In this retrospective diagnostic study, patients undergoing revision surgery after total hip or knee arthroplasty (n = 119) between January 2015 and July 2018 were included. Multiple specimens of the periprosthetic membrane and pseudocapsule were obtained for histopathological analysis at revision arthroplasty. Based on the Infectious Diseases Society of America (IDSA) 2013 criteria, the International Consensus Meeting (ICM) 2018 criteria, and the European Bone and Joint Infection Society (EBJIS) 2021 criteria, PJI was defined. Using a mixed effects logistic regression model, the sensitivity and specificity of the histological diagnosis were calculated. The optimal number of periprosthetic tissue specimens for histopathological analysis was determined by applying the Youden index. Based on the EBJIS criteria (excluding histology), 46 (39%) patients were classified as infected. Four to six specimens showed the highest Youden index (four specimens: 0.631; five: 0.634; six: 0.632). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of five tissue specimens were 76.5% (95% confidence interval (CI) 67.6 to 81.4), 86.8% (95% CI 81.3 to 93.5), 66.0% (95% CI 53.2 to 78.7), and 84.3% (95% CI 79.4 to 89.3), respectively. The area under the curve (AUC) was calculated with 0.81 (as a function of the number of tissue specimens). Applying the ICM and IDSA criteria (excluding histology), 40 (34%) and 32 (27%) patients were categorized as septic. Three to five specimens had the highest Youden index (ICM 3: 0.648; 4: 0.651; 5: 0.649) (IDSA 3: 0.627; 4: 0.629; 5: 0.625). Three to six tissue specimens of the periprosthetic membrane and pseudocapsule should be collected at revision arthroplasty and analyzed by a pathologist experienced and skilled in interpreting periprosthetic tissue.Cite this article: Bone Joint J2023;105-B(2):158-165.

Complex diagnostic study of twenty-nine patients revised for periprosthetic hip joint reinfection and the role of the histological method in detection of osteomyelitis

Abstract. Introduction Re-infection rates remain high after revision for periprosthtic joint infection (PJI). We conducted a complex diagnostic study of hip PJI and studied bone tissue involvement into the infectious process. Material and methods Twenty-nine patients treated for PJI were examined. Ten patients had acute, seven had late and 12 hematogenous infection (Tsukayama, 1996). Clinical, laboratory, microbiological, radiographic and histological methods were used for PJI detection. Results ESR exceeded the threshold in 16 and CRP in 23 cases. Pathogenic microorganisms were confirmed in 23 cases. Radiographic manifestations of periprosthetic bone destruction were seen in 14 patients with late and hematogenous infection. The histological study showed signs of osteomyelitis in 19 patients, including five cases with acute PJI. Histological study of periprosthetic and pseudosynovial membranes revealed PJI in 20 patients. The histological and microbiological tests together confirmed PJI in 27 cases (92 %). ROC analysis showed that the accuracy of CRP in the diagnosis of osteomyelitis exceeded that of ESR due to its higher sensitivity. Microbiological tests showed satisfactory sensitivity but low specificity. The radiographic study had an extremely low sensitivity and low specificity. Histology of membranes was quite sensitive and specific for the diagnosis of osteomyelitis but did not reach the level of bone histology. Conclusions Our complex diagnostic study has enabled to accurately characterize the septic process in revision arthroplasty. The histological findings show that osteomyelitis of different severity might develop in each PJI type. This fact may be a key factor for a surgeon in choosing a more reliable treatment option.

Histological evaluation of periprosthetic infection using HOES scale and CD15 expression analysis at the stage of the hip revision arthroplasty

Background.The effectiveness improvement and standardization of the methods of histological diagnosing periprosthetic infection (PPI) is an urgent task in the treatment of complications after large joint arthroplasty. Purpose of the study— Histopathological evaluation of the infection involvement of periprosthetic tissues at the stage of revision arthroplasty for deep infection of the hip using HOES scale and immunohistochemical analysis of CD15 expression.Materials and Methods.A single-center prospective study was performed on the clinical intraoperative material obtained at the stage of revision arthroplasty of the hip in 27 patients at the age of 65 (55÷69) years. The group of examination included patients with acute and chronic forms of deep periprosthetic infection. Light-optical microscopic investigation of the samples of periprosthetic connective-tissue membrane and bone tissue from the foci of infectious involvement was made on paraffin sections stained with hematoxylin and eosin; with the immunohistochemical reaction to determine the expression of CD15 neutrophil granulocyte markers. HOES Scale for pathohistological assessment was used in order to objectify osteomyelitis signs in periprosthetic bone tissue.Results. The signs of acute and chronic stages of periprosthetic osteomyelitis were observed in 9/16 patients with PPI chronic course within 1–30 months of postoperative period, from one to 18 months after manifestation of the symptoms. The signs of subsided osteomyelitis were determined in 12/27 patients with PPI of acute and chronic forms. Infected periprosthetic membranes were found in 19/27 clinical cases in the early and longterm time periods after arthroplasty surgery. A direct significant correlation was revealed between histopathological signs of infecting the periprosthetic bone and the connective-tissue periprosthetic membrane, especially strong one in patients with acute and chronic PPI osteomyelitis.Conclusion. The use of HOES Scale and the analysis of CD15 expression ensure the objectivity of PPI histological diagnosing. The results obtained indicate an increased risk of osteomyelitis development in patients with chronic periprosthetic infection after the hip arthroplasty.

Naringin increases osteoprotegerin expression in fibroblasts from periprosthetic membrane by the Wnt/\u03b2-catenin signaling pathway

BackgroundThe osteoclast bone resorption is critical in aseptic loosening after joint replacement. The balance between activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) is considered to play a central role in osteoclast maturation. Fibroblasts from the periprosthetic membrane express RANKL and promote osteoclast formation. Studies have demonstrated that naringin inhibited osteoclastogenesis and wear particle-induced osteolysis. In this study, the naringin-induced OPG/RANKL effects and its underlying mechanism were studied in fibroblasts from periprosthetic membrane.MethodsFibroblasts were isolated from the periprosthetic membrane during hip arthroplasty for revision due to aseptic loosening. Fibroblasts were cultured and treated with or without naringin and DKK-1 (the classical inhibitor of Wnt/β-catenin signaling pathway). OPG and RANKL mRNA and protein levels, gene expression of β-catenin, and cyclin D1, which participate in the Wnt signaling pathway, were examined by real-time polymerase chain reaction and enzyme-linked immunosorbent assay.ResultsThe mRNA and protein levels of OPG were enhanced by naringin in a dose-dependent manner compared to that of the non-treated control. In contrast, naringin did not affect the expression of RANKL. Importantly, DKK-1 attenuated OPG expression in fibroblasts under naringin treatment. Moreover, naringin stimulated the gene expression of β-catenin and cyclin D1 in fibroblasts, and the effect could be inhibited by DKK-1.ConclusionThe results indicated that naringin enhanced OPG expression through Wnt/β-catenin signaling pathway in fibroblasts from periprosthetic membrane, which may be useful to inhibit periprosthetic osteolysis during aseptic loosening after total joint arthroplasty.

Noninvasive diagnostic of periprosthetic joint infection by urinary peptide markers: A preliminary study

Previous immunohistochemical analyses revealed altered protein expression in the periprosthetic membranes of patients with periprosthetic joint infection (PJI). Proteins are degraded to peptides that may pass the blood-kidney barrier depending on their size. The aim of this study was to evaluate if PJI can be diagnosed based on the urinary peptide excretion pattern. Thirty patients undergoing removal of their hip or knee prostheses due to septic or aseptic loosening were enrolled. Specimen sampling was performed according to the MusculoSkeletal Infection Society criteria. A urinary sample was analyzed before surgery using capillary electrophoresis coupled with mass spectrometry. Peptides with differential urinary excretion between groups were used to establish a multimarker model. A total of 137 peptides were differentially excreted between the septic and aseptic groups. The majority of identified peptide markers were fragments of structural extracellular matrix proteins, potentially due to their origin from the periprosthetic membrane. A marker model with 83 peptides achieved the best diagnostic performance for diagnosing PJI with a sensitivity of 95%, a specificity of 90%, and an area under the curve of 0.96. The altered protein expression pattern in the periprosthetic membrane of PJI is most likely reflected in excreted urinary peptides. Thus, the diagnosis of PJI by urinary peptide markers seems to be a reasonable approach.

Failed Total Hip Arthroplasty: Diagnostic Performance of Conventional MRI Features and Locoregional Lymphadenopathy to Identify Infected Implants.

Very little has been published about the diagnostic performance of MRI in total hip arthroplasty (THA) infection. To determine the diagnostic performance of conventional MRI features and of new lymph nodal indices to identify infection in patients with failed THA. Retrospective. In all, 119 patients (66 females; age 66.9 ± 12.4 years) with failed THA. Metal artifact reduction sequence (MARS) protocol including short tau inversion recovery and turbo spin-echo T1 - and T2 -weighted sequences at 1.5T. Patients underwent pelvis MRI prior to failed THA revision. MRIs were reviewed to identify periprosthetic bone destruction, soft-tissue mass, effusion, synovitis, lamellated synovitis, extracapsular edema, fibrous periprosthetic membrane, bone edema, and extracapsular collection/sinus tract. The number and maximum diameter of inguinal, obturator and iliac lymph nodes of the affected hip were assessed and normalized to those of the unaffected hip to calculate the ratio of nodal size (RNS), ratio of node number (RNN), difference of nodal size (DNS), and difference of node number (DNN). The Mann-Whitney U-and chi-square test were used. Diagnostic performance of indices and odds ratios (OR) were calculated. RNS, RNN, DNS, and DNN indices were significantly different (P = 0.000) between infected and noninfected THA, with accuracies ranging from 84.8% (RNS) and 93.1% (RNN). All other MRI features were significantly more prevalent in infected THA (P ≤ 0.002), except bone destruction, periarticular soft-tissue mass, and fibrous membrane (P ≥ 0.031). Sensitivities ranged from 7.9% (soft-tissue mass) to 76.3% (effusion/bone edema), specificity from 45.7% (bone destruction) to 97.5% (synovitis/lamellated synovitis), accuracy from 49.6% (bone destruction) to 81.5% (synovitis), OR from 0.261 (soft-tissue mass) to 35.550 (synovitis). Conventional MRI features have limited accuracy to differentiate septic and aseptic THA failure. Lymph nodal indices, particularly those related to nodal number, may represent biomarkers of THA infection. 3 TECHNICAL EFFICACY STAGE: 2.

Naringin inhibits titanium particles-induced up-regulation of TNF-α and IL-6 via the p38 MAPK pathway in fibroblasts from hip periprosthetic membrane

ABSTRACT Aims Inflammatory responses to wear debris cause osteolysis that leads to aseptic loosening and hip arthroplasty failure. Wear debris stimulate macrophages and fibroblasts to secret proinflammatory cytokines, including TNF-α and IL-6, which have been specifically implicated in periprosthetic osteolysis and osteoclast differentiation. Naringin has anti-inflammatory effect in macrophages. Moreover, naringin inhibited osteoclastogenesis and wear particles-induced osteolysis. In this study, we examined the potential inhibitory effects of naringin on titanium (Ti) particle-induced proinflammatory cytokines secretion in fibroblasts and the possible underlying molecular mechanisms. Materials and methods Fibroblasts were isolated from periprosthetic membrane at the time of revision surgery performed due to aseptic loosening after hip arthroplasty and were cultured in the presence or absence of Ti particles, naringin and mitogen-activated protein kinase (MAPK) inhibitors, PD98059 (a selective inhibitor of ERK), SP600125 (a selective inhibitor of JNK), and SB203580 (a selective inhibitor of p38). TNF-α and IL-6 assays were performed using enzyme-linked immunosorbent assay kits. The phosphorylation levels of p38 and nuclear factor kappa B p65 (NF-κB p65) were examined by western blot. Results Naringin or SB203580 pretreatment significantly suppressed the secretion of TNF-α and IL-6 induced by titanium particles in fibroblasts, while inhibition of ERK or JNK pathways showed no effect on production of TNF-α and IL-6. Moreover, naringin inhibited Ti particle-induced phosphorylation of p38 and p65. Conclusions These results indicated that naringin could inhibit Ti particle-induced inflammation in fibroblasts by inhibiting p38 MAPK/NF-κB p65 activity and might be a potential drug for the treatment of inflammatory periprosthetic osteolysis after arthroplasty.

Safety assessment of microsilver-loaded poly(methyl methacrylate) (PMMA) cement spacers in patients with prosthetic hip infections: Results of a prospective cohort study.

ObjectivesPreclinical data showed poly(methyl methacrylate) (PMMA) loaded with microsilver to be effective against a variety of bacteria. The purpose of this study was to assess patient safety of PMMA spacers with microsilver in prosthetic hip infections in a prospective cohort study.MethodsA total of 12 patients with prosthetic hip infections were included for a three-stage revision procedure. All patients received either a gentamicin-PMMA spacer (80 g to 160 g PMMA depending on hip joint dimension) with additional loading of 1% (w/w) of microsilver (0.8 g to 1.6 g per spacer) at surgery 1 followed by a gentamicin-PMMA spacer without microsilver at surgery 2 or vice versa. Implantation of the revision prosthesis was carried out at surgery 3.ResultsIn total, 11 of the 12 patients completed the study. No argyria or considerable differences in laboratory parameters were detected. Silver blood concentrations were below or around the detection limit of 1 ppb in ten of the 11 patients. A maximum of 5.6 ppb at 48 hours after implantation of the silver spacer, which is below the recommended maximum level of 10 ppb, was found in one patient. No silver was detected in the urine. Drainage fluids showed concentrations between 16.1 ppb and 23.3 ppb at 12 hours after implantation of the silver spacers, and between 16.8 ppb to 25.1 ppb at 48 hours after implantation. Pathohistological assessment of the periprosthetic membrane did not reveal any differences between the two groups.ConclusionMicrosilver-loaded gentamicin-PMMA spacers showed good biocompatibility and the broad antimicrobial activity warrants further clinical research to assess its effectivity in reducing infection rates in prosthetic joint infection.Cite this article: V. Alt, M. Rupp, K. Lemberger, T. Bechert, T. Konradt, P. Steinrücke, R. Schnettler, S. Söder, R. Ascherl. Safety assessment of microsilver-loaded poly(methyl methacrylate) (PMMA) cement spacers in patients with prosthetic hip infections: Results of a prospective cohort study. Bone Joint Res 2019;8:387–396. DOI: 10.1302/2046-3758.88.BJR-2018-0270.R1.

Role of Brd4 in the production of inflammatory cytokines in mouse macrophages treated with titanium particles.

Brd4 protein is an important epigenetic regulator involved in the process of inflammatory cytokine production in many diseases. However, whether and how Brd4 participates in the process of wear-particle-induced inflammation remain unclear. This study aimed to investigate the potential role of Brd4 in titanium (Ti) particle-induced inflammatory cytokine production in mouse macrophage RAW264.7 cells. Our experiment detected Brd4 expressed in both normal synovium and periprosthetic osteolysis interface membrane, but the expression increased in the interface membrane as compared with that in normal synovium. Treatment with Ti particles significantly increased TNF-α, IL-6, and IL-1β production in RAW264.7 cells, which was inhibited by JQ1 or Brd4-siRNA. Ti particles enhanced the expression of Brd4, which was abrogated by JQ1. Ti particles enhanced NF-κB p65 and IKK phosphorylation and attenuated IκBα protein expression, which were abrogated by JQ1. Co-immunoprecipitation analysis indicated that Ti particles promoted the binding of Brd4 to acetylated NF-κB p65 (lysine-310), which was also abrogated in JQ1-treated RAW264.7 cells. In conclusion, Brd4 expression increases in interface membrane and Brd4 participates in the production of pro-inflammatory cytokines induced by Ti particles via promoting the activation of NF-κB signaling and binding to acetylated NF-κB p65 (lysine-310) in mouse macrophages.

Water Jet Applicator for Interface Tissue Removal in Minimally Invasive Hip Refixation: Testing the Principle and Design of Prototype

Mechanical loosening of implants is in the majority accompanied with a periprosthetic interface membrane, which has to be removed during revision surgery. The same is true if a minimal invasive (percutaneous) refixation of a loose implant is done. We describe the requirements for a waterjet applicator for interface tissue removal for this percutaneous hip refixation technique. The technical requirements were either obtained from a literature review, a theoretical analysis, or by experimental setup. Based on the requirements, a waterjet applicator is designed which is basically a flexible tube (outer diameter 3 mm) with two channels. One channel for the water supply (diameter 0.9 mm) and one for suction to evacuate water and morcellated interface tissue from the periprosthetic cavity. The applicator has a rigid tip (length 6 mm), which directs the water flow to create two waterjets (diameter 0.2 mm), both focused into the suction channel. The functionality of this new applicator is demonstrated by testing a prototype of the applicator tip in an in vitro experimental setup. This testing has shown that the designed applicator for interface tissue removal will eliminate the risk of water pressure buildup; the ejected water was immediately evacuated from the periprosthetic cavity. Blocking of the suction opening was prevented because the jets cut through interface tissue that gets in front of the suction channel. Although further development of the water applicator is necessary, the presented design of the applicator is suitable for interface tissue removal in a minimally invasive hip refixation procedure.

Fluorescence in Situ Hybridization (FISH) for the Diagnosis of Periprosthetic Joint Infection in Formalin-Fixed Paraffin-Embedded Surgical Tissues.

As the number of arthroplasties performed increases, periprosthetic joint infection (PJI) represents a common and challenging problem. The Musculoskeletal Infection Society (MSIS) recommends diagnosing PJI according to its guidelines. The aim of the current study was to assess whether fluorescence in situ hybridization (FISH) analysis of formalin-fixed paraffin-embedded periprosthetic membranes can successfully improve the diagnosis of infection in patients with orthopaedic implants. We retrospectively analyzed 88 periprosthetic membranes of joint prostheses using FISH analysis according to a standard protocol, with a probe targeting a sequence found in most bacteria. We compared the results with routine clinical classification according to the guidelines of the MSIS, microbiological culture, and histopathological classification according to Morawietz and Krenn. We additionally performed FISH analysis using 2 species-specific probes for several culture-positive cases. FISH successfully detected bacteria in 38 (95%) of 40 periprosthetic membranes that were rated positive by clinical classification. FISH results compared with clinical classification demonstrated a sensitivity of 95% (95% confidence interval [CI], 83.08% to 99.39%), a specificity of 85.42% (95% CI, 72.24% to 93.93%), a positive predictive value of 84.44% (95% CI, 70.55% to 93.50%), and a negative predictive value of 95.35% (95% CI, 84.19% to 99.43%). FISH results compared with histopathological classification demonstrated a sensitivity of 95.12% (95% CI, 83.47% to 99.40%), a specificity of 87.23% (95% CI, 74.26% to 95.17%), a positive predictive value of 86.67% (95% CI, 73.21% to 94.95%), and a negative predictive value of 95.35% (95% CI, 84.19% to 99.43%). We successfully detected Pseudomonas aeruginosa and Staphylococcus aureus with species-specific FISH probes in all cases that were positive for these respective bacteria by microbiological culture. FISH-based diagnosis of PJI is feasible and can be used as an additional diagnostic criterion. FISH not only can detect bacteria in periprosthetic membranes but can also differentiate pathogens at the species level. FISH represents a fast and reliable tool for detecting PJI in periprosthetic membranes, especially in combination with clinical and histopathological classification. Diagnostic Level III. See Instructions for Authors for a complete description of levels of evidence.

Usefulness of Culturing the Periprosthetic Membrane or Neosynovium for the Diagnosis of Infection During Hip and Knee Revision Arthroplasty

Identification of microorganisms is critical for correct management of an infected arthroplasty. Our hypothesis is that the culture yield depends on the location around the prosthesis from which samples are obtained. This prospective study included 298 revisions of the hip (123) and knee (175). We compared the yield of the intraoperative samples obtained, which included synovial fluid (two), neosynovium (two), and periprosthetic membrane (two). Cultures were positive in 28 cases, in which 15 had the same diagnosis considering either the neosynovium or the membrane, and there were 3 cases in which the infection could have been diagnosed only by considering the combination of both. In all, there were 8 cases in which the infection might have been misdiagnosed unless considering a combination of both solid tissue samples (P = 0.004). The yields of the periprosthetic membrane and neosynovium do not differ significantly, and we recommend considering a combination of both. Diagnostic Level II.

Antibiotics release from cement spacers used for two-stage treatment of implant-associated infections after total joint arthroplasty.

Two‐stage revision arthroplasty is the treatment of choice for periprosthetic infection, a serious complication after knee or hip arthroplasty. Our prospective clinical trial aimed to investigate the concentrations of gentamicin and vancomycin in wound exudate and tissue in two‐stage revision arthroplasty. Wound exudate and periprosthetic membrane samples were collected from 18 patients (10 hip and eight knee patients), who were due for two‐stage treatment after a periprosthetic joint infection. Samples were taken during insertion of antibiotic‐impregnated spacers and after their removal. The concentrations of gentamicin and vancomycin in wound exudates and adjacent tissue were analyzed using high‐performance liquid chromatography mass spectrometry. Average time period of spacer implantation was 13.6 weeks (9.3–22.6 weeks). The concentration of vancomycin in wound exudate decreased from a median of 43.28 μg/mL (0.28–261.22) after implantation to 0.46 μg/mL (0.13–37.47) after the removal of the spacer. In the adjacent tissue, vancomycin concentration was mainly undetectable prior to spacer implantation (0.003 μg/g [0.003–0.261]) and increased to 0.318 μg/g [0.024–484.16] at the time of spacer removal. This was also observed for gentamicin in the tissue of patients who previously had cement‐free implants (0.008 μg/g [0.008–0.087] vs. 0.164 μg/g [0.048–71.75]) while in the tissue of patients with previously cemented prosthesis, baseline concentration was already high (8.451 μg/g [0.152–42.926]). Despite the rapid decrease in antibiotics release from spacer cement observed in vitro, in vivo antibiotics are much longer detectable, especially in the adjacent soft tissue. © 2018 The Authors. Journal of Biomedical Materials Research Part B: Applied Biomaterials Published By Wiley Periodicals, Inc. J Biomed Mater Res B Part B, 2019. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1587–1597, 2019.

Diagnostic guidelines for the histological particle algorithm in the periprosthetic neo-synovial tissue

BackgroundThe identification of implant wear particles and non-implant related particles and the characterization of the inflammatory responses in the periprosthetic neo-synovial membrane, bone, and the synovial-like interface membrane (SLIM) play an important role for the evaluation of clinical outcome, correlation with radiological and implant retrieval studies, and understanding of the biological pathways contributing to implant failures in joint arthroplasty. The purpose of this study is to present a comprehensive histological particle algorithm (HPA) as a practical guide to particle identification at routine light microscopy examination.MethodsThe cases used for particle analysis were selected retrospectively from the archives of two institutions and were representative of the implant wear and non-implant related particle spectrum. All particle categories were described according to their size, shape, colour and properties observed at light microscopy, under polarized light, and after histochemical stains when necessary. A unified range of particle size, defined as a measure of length only, is proposed for the wear particles with five classes for polyethylene (PE) particles and four classes for conventional and corrosion metallic particles and ceramic particles.ResultsAll implant wear and non-implant related particles were described and illustrated in detail by category. A particle scoring system for the periprosthetic tissue/SLIM is proposed as follows: 1) Wear particle identification at light microscopy with a two-step analysis at low (× 25, × 40, and × 100) and high magnification (× 200 and × 400); 2) Identification of the predominant wear particle type with size determination; 3) The presence of non-implant related endogenous and/or foreign particles. A guide for a comprehensive pathology report is also provided with sections for macroscopic and microscopic description, and diagnosis.ConclusionsThe HPA should be considered a standard for the histological analysis of periprosthetic neo-synovial membrane, bone, and SLIM. It provides a basic, standardized tool for the identification of implant wear and non-implant related particles at routine light microscopy examination and aims at reducing intra-observer and inter-observer variability to provide a common platform for multicentric implant retrieval/radiological/histological studies and valuable data for the risk assessment of implant performance for regional and national implant registries and government agencies.

Protein phosphatase 2A as a new target for downregulating osteoclastogenesis and alleviating titanium particle-induced bone resorption

Receptor activator of nuclear factor-кB ligand (RANKL)-induced osteoclastogenesis is believed to play a critical role in osteolytic diseases including peri-prosthetic osteolysis (PPO), the primary reason for implant failure and revision surgery. In this study, we observed that protein phosphatase 2A (PP2A), a major serine-threonine phosphatase, was highly expressed in human periprosthetic interface membranes with aseptic loosening and in a murine osteolysis model induced by titanium particle irritation. PP2A inhibition effectively alleviated titanium particle-induced bone destruction at osteolytic sites. In addition, PP2A downregulation significantly decreased osteoclast numbers and RANKL expression, compared with in animals treated with only titanium. Mechanistically, a PP2A selective inhibitor or PP2A siRNA suppressed osteoclastogenesis and alleviated osteoclastic resorption by inhibiting the RANKL-induced nuclear factor-кB and c-Jun N-terminal kinase signaling pathways. Downstream NFATc1 and c-Fos expression were also substantially suppressed by PP2A inhibition or knockdown. Our findings support the importance of PP2A during osteoclastogenesis, identifying PP2A as a novel target for treating particle-induced or other osteoclast-mediated bone resorption diseases. Statement of significanceExcessive osteoclast activation disrupts bone homeostasis and leads to osteoclast-mediated bone resorption diseases, such as peri-prosthetic osteolysis, regarded as the primary reason for implant failure and revision surgery. Here, we firstly demonstrated protein phosphatase 2A (PP2A), a major serine-threonine phosphatase, was highly expressed in human periprosthetic interface membranes with aseptic loosening and murine osteolysis model. Moreover, PP2A inhibition effectively alleviated titanium particle-induced bone destruction and decreased osteoclast numbers. Meanwhile, a PP2A selective inhibitor or PP2A siRNA suppressed osteoclastogenesis and alleviated osteoclastic resorption by inhibiting the nuclear factor-кB and c-Jun N-terminal kinase signaling pathways. Thus, PP2A is involved in osteoclastogenesis and could be a promising target for regulating bone homeostasis and osteolytic responses.