Synovial fluid NMR-based metabolomics in septic and aseptic revision total knee arthroplasty: Implications on diagnosis and treatment.

Periprosthetic joint infection (PJI) is one of the most challenging complications following total knee arthroplasty. Despite its importance, there is a paucity of reports in the literature regarding its pathogenesis. Recently, cellular metabolic reprogramming has been shown to play an important role in the progression and outcome of infectious diseases. Therefore, the aim of this study was to evaluate the metabolites composition of the synovial fluid from patients with PJI or aseptic failure of total knee arthroplasties. The synovial fluids from 21 patients scheduled for revision total knee arthroplasty (11 with the diagnosis of PJI and 10 with aseptic failures) were analyzed using 1D 1H NMR spectroscopy. Univariate and multivariate statistical analyzes were used to identify metabolites that were differentially abundant between those groups. A total of 28 metabolites were identified and five of them found to be differentially abundant between infected and non-infected synovial fluids. Lactate, acetate and 3-hydroxybutyrate were found to be in a higher concentration, and glucose and creatine were found reduced in the synovial fluid from PJI patients. Synovial fluid from patients with PJI exhibit a distinct metabolic profile, possibly reflecting metabolic adaptation that occurs in the infected periprosthetic microenvironment. Further research and studies are warranted to gain a broader insight into the metabolic pathways engaged by both pathogen and immune cells in the context of a PJI.

An update on the study of synovial fluid in the geriatric patient.

BACKGROUND: The characteristics of synovial fluid (SF) in geriatric patients differ from those in younger patients. In Mexico, epidemiologic data on the incidence of different rheumatic diseases in geriatric patients are scarce. OBJECTIVE: To describe the physical characteristics of geriatric SF and the prevalence of crystals in knee and other joint aspirates from patients with previously diagnosed joint disease. MATERIALS AND METHODS: A retrospective study was performed with a baseline of 517 SF samples between 2011 and 2023. White blood cell count was performed by Neubauer chamber and crystals were identified by polarized light microscopy. Descriptive statistical analysis was performed and prevalence was reported as a percentage. RESULTS: The mean age of the adults was 73.5±5.0 years, 54.4% were women and 45.6% were men. The mean SF volume was 6.3±9.5mL in older adults and 15.3±24.9mL in those younger than 65 years. The mean viscosity in older adults was 9.5±4.5mm and the mean leukocyte count was 7352±16,402leukocytes/mm3. Seventy percent of the older adults' SFs were referred to the laboratory for osteoarthritis (OA), with lower proportions for rheumatoid arthritis (RA) (14.6%) and gout (5.1%). Of the crystals observed in the geriatric population, 14.6% corresponded to monosodium urate crystals (CUM) and 18.9% to calcium pyrophosphate crystals (CPP). CONCLUSIONS: The characteristics of LS in older adults were smaller volume, increased viscosity, and non-inflammatory. The main diagnoses were OA, RA, and gout. The crystal content of the SF of the geriatric population corresponded mainly to CPP.

Neutrophil Extracellular Trap-related Biomarkers Are Increased in the Synovial Fluid of Patients With Periprosthetic Joint Infections.

BACKGROUND: The diagnosis of periprosthetic joint infection (PJI) is a major challenge in clinical practice. The role of neutrophils in fighting infection has been increasingly understood, and one mechanism of action of these cells is neutrophil extracellular traps. However, little is known about this process in PJI. QUESTIONS/PURPOSES: (1) Are the biomarkers of neutrophil extracellular trap formation (citrullinated histone H3 [H3Cit], cell-free DNA [cf-DNA], and myeloperoxidase [MPO]) increased in the synovial fluid of patients with PJI? (2) What is the diagnostic accuracy of biomarkers of neutrophil extracellular trap formation for PJI? METHODS: Between May 2020 and March 2021, 43 patients who underwent revision THA or TKA were enrolled in this study. Eleven patients were excluded and 32 patients were categorized into the PJI group (n = 16) or non-PJI group (n = 16) according to the 2018 Second International Consensus Meeting on Musculoskeletal Infection criteria. There were 15 men and 17 women in this study, with a median (range) age of 70 years (60 to 80 years). Twenty-seven patients had TKA and five had THA. We measured cf-DNA, MPO, and H3Cit in synovial fluid. The sensitivity, specificity, and receiver operating characteristic curve were calculated for each biomarker using the Musculoskeletal Infection Society criteria as the gold standard for diagnosis and considering a clinical surveillance of 2 years for patients in the non-PJI group. RESULTS: Patients with PJI had higher levels of synovial fluid cf-DNA (median [range] 130 ng/µL [18 to 179] versus 2 ng/µL [0 to 6]; p < 0.001), MPO (1436 ng/µL [55 to 3996] versus 0 ng/µL [0 to 393]; p < 0.001), and H3Cit (2115 ng/µL [5 to 2885] versus 3 ng/µL [0 to 87]; p < 0.001) than those in the non-PJI group. In receiver operating characteristic curve analyses, we observed near-perfect performance for all biomarkers evaluated, with an area under the curve of 1 (95% CI 0.9 to 1), 0.98 (95% CI 0.9 to 1), and 0.94 (95% CI 0.8 to 0.99) for cf-DNA, MPO, and H3Cit, respectively. The sensitivity for detecting PJI using synovial fluid was 100% for cf-DNA, 94% for MPO, and 88% for H3Cit. The specificity was 100% for cf-DNA and MPO, and 88% for H3Cit. CONCLUSION: Our results show that neutrophils in the periprosthetic microenvironment release neutrophil extracellular traps as part of the bactericidal arsenal to fight infection. These results allow a better understanding of the cellular and molecular processes that occur in this microenvironment, enabling the design of more assertive strategies for identifying new biomarkers and improving the available ones. Novel studies are needed to define whether and how neutrophil extracellular trap-related biomarkers can be useful for diagnosing PJI. LEVEL OF EVIDENCE: Level II, diagnostic study.

Cell-free Deoxyribonucleic Acid: A Potential Biomarker of Chronic Periprosthetic Knee Joint Infection.

BACKGROUND: The correct diagnosis of a chronic periprosthetic joint infection (PJI) is a major challenge in clinical practice, with the "gold standard" for diagnosis yet to be established. Synovial fluid analysis has been proven to be a useful tool for that purpose. Cell-free DNA (cf-DNA) levels have been shown to be increased in several conditions such as cancer, trauma, and sepsis. Therefore, this study was designed to evaluate the potential of synovial fluid cf-DNA quantification for the diagnosis of chronic periprosthetic infections following total knee arthroplasty. METHODS: A prospective study with patients undergoing total knee arthroplasty revision surgery for any indication was performed. PJI diagnosis was defined according to the Second International Consensus Meeting on Musculoskeletal Infection (2018) criteria. The study cohort consisted of 26 patients classified as infected and 40 as noninfected. Synovial fluid cf-DNA direct quantification by fluorescent staining was made. Sensitivity, specificity, and receiver operating characteristic curve were calculated. RESULTS: The cf-DNA levels were significantly higher in patients who had PJIs (122.5 ± 57.2 versus 4.6 ± 2.8 ng/µL, P < .0001). With a cutoff of 15 ng/µL, the area under the receiver operating characteristic, sensitivity, and specificity of cf-DNA were 0.978, 96.2%, and 100%, respectively. CONCLUSION: The present study has shown that cf-DNA is increased in synovial fluid of patients who have chronic PJIs. It is a promising biomarker for knee PJI diagnosis and further studies are needed to confirm its utility.

Biomarcadores inflamatorios en líquido sinovial de trastornos intraarticulares de la articulación temporomandibular. Revisión sistemática / Inflammatory biomarkers in synovial fluid of intra-articular disorders of the temporomandibular joint. Systematic review

RESUMEN Objetivo: Determinar los biomarcadores inflamatorios del líquido sinovial (LS) de pacientes adultos con trastornos intraarticulares (TI) de la articulación temporomandibular (ATM) y su capacidad diagnóstica. Métodos: Se realizó búsqueda electrónica/manual de artículos (2010-2019) en paralelo por dos investigadores. La calidad de los estudios, se determinó por medio de CONSORT y STROBE y el sesgo según criterios Cochrane RoB 2 en ensayos clínicos aleatorizados y Escala Newcastle-Ottawa en estudios observacionales. Se estudiaron pacientes con TI de la ATM y determinación de biomarcadores del LS. Resultados: De 264 artículos encontrados, 6 cumplieron los criterios inclusión-exclusión, incluyendo 262 pacientes, [OA=153, 93 con desplazamientos discales (DD) y 16 con OA+DD]. Todas las muestras fueron obtenidas por artrocentesis y detectadas por ELISA. Se determinaron 19 biomarcadores en pacientes con OA; 9 en DD y 2 en diagnosticados con OA+DD. El incremento de biomarcadores en el LS de la ATM se asocia con TI. Conclusión: Los biomarcadores detectados con mayor frecuencia en LS de pacientes con TI de ATM fueron IL-1β, IL-6 y TNF-α y en segunda frecuencia TGF-β1, MMP-3 e IFN-γ. Dada la inconsistencia de los protocolos utilizados la evidencia fue débil, imposibilitando asociar biomarcadores con diagnóstico de TI determinado, ni efectuar análisis estadístico.

ABSTRACT: Objective: To determine the evidence of inflammatory biomarkers present in the synovial fluid (SF) of adult patients with intra-articular disorders (ID) of the temporomandibular joint (TMJ) and their diagnostic ability. Methods: Electronic/manual search of articles (2010-2019) was performed. Data were extracted in duplicate. The quality of the studies was determined by CONSORT, STROBE and risk of bias was determined by Cochrane RoB 2 and Newcastle-Ottawa Scale. The populations studied were patients with TMJ ID and with studies of SF biomarkers. Results: Out of 264 articles found, 6 met the inclusion-exclusion criteria, including 262 patients, 93 with disc displacements (DD) and 16 with OA+DD. All samples were obtained by arthrocentesis and detected by ELISA. Nineteen biomarkers were evaluated in patients with OA, 9 in patients with DD and 2 in those diagnosed with OA+DD. Increased inflammatory biomarkers in the SF of TMJ are associated with ID. Conclusion: The most frequent biomarkers detected in SF of patients with TMJ ID were IL-1β, IL-6 and TNF-α and in second frequency TGF-β1, MMP-3 and IFN-γ. Given the inconsistency of the protocols used, the evidence was weak, making it impossible to associate biomarkers with a given IT diagnosis, or to perform statistical analysis.

Alpha-Defensin Offers Limited Utility in Routine Workup of Periprosthetic Joint Infection.

BACKGROUND: Alpha-defensin (AD) is a synovial biomarker included as a minor criterion in the scoring system for diagnosing periprosthetic joint infection (PJI). The purpose of this study is to study the impact of AD on diagnosis and management of PJI. METHODS: Synovial fluid from 522 patients after total knee and hip arthroplasty was retrospective reviewed. Synovial white blood cell count, percentage of neutrophils, and culture from the AD immunoassay laboratory were reviewed with serum erythrocyte sedimentation rate and C-reactive protein values from our institution. A modified version of the 2018 scoring system for diagnosis of PJI was used, only scoring white blood cell count, percentage of neutrophils, erythrocyte sedimentation rate, and C-reactive protein. AD was then analyzed with these scores to determine if AD changed diagnostic findings or clinical management. RESULTS: Eight-two patients were categorized as "infected" (score ≥6), of which 76 patients had positive AD. Of the 6 "infected" patients with negative AD, 2 had positive cultures (Staphylococcus epidermidis). Two-hundred thirteen patients were diagnosed as "possibly infected" (score 2-5). Fourteen of these patients had positive AD, of which 5 had positive cultures assisting with the diagnosis. The AD test changed the diagnosis from "possibly infected" to "infected" in 8 patients (1.5%) but only altered treatment plan in 6 patients (1.1%). A score <2 (not infected) was calculated in 227 patients with no patients having positive AD. CONCLUSION: AD may be beneficial in some cases where laboratory values are otherwise equivocal; however, its routine use for the diagnosis of PJI may not be warranted.

Evaluation of biomarkers of joint damage in patients subjected to arthroscopy.

AIMS: Recently, the determination of biochemical markers has been intensely explored to better understand the mechanisms underlying knee OA. In this study, we aimed to explore the expression pattern of five biochemical markers in patients with knee OA. METHODS: After IRB approval and signed informed consent, 26 patients were enrolled. Serum and synovial samples were collected prior to knee arthroscopy. Pre-operative assessment included diagnosis, Lysholm, Tegner Activity Scale, IKDC score, and radiographic Kellgren and Lawrence classification. ELISA of CTX-I, CTX-II, NTX-I, MMP3, and MMP13 were measured in serum and synovial fluid samples. RESULTS: Twenty-six patients were included, with a mean age of 42 ± 15 years old. Mean results and standard deviation of the biomarkers in serum were as follows: CTX-I 5.8 ± 5.5 ng/mL, CTX-II 3.8 ± 1.7 ng/mL, NTX-I 52 ± 71 (nM BCE), MMP3 1.18 ± 0.6 ng/mL, and MMP13 1243.6 ± 1422 pg/mL; synovial fluid results were as follows: CTX-I 0.74 ± 0.5 ng/mL, CTX-II 5.1 ± 2.5 ng/mL, NTX-I 254 ± 85 (nM BCE), MMP3 0.4 ± 0.4 ng/mL, and MMP13 797 ± 1391 pg/mL. We observed a differential pattern of expression in serum NTX-I in patients with chronic meniscus injuries when compared with ACL injuries or cartilage lesions. CONCLUSIONS: In conclusion, the clinical criteria of early OA are useful to categorize patients with knee conditions. The biochemical markers explored did not yield a differential pattern that can be associated with this classification. Serum NTX-I could be a useful marker of chronic meniscal lesion in future longitudinal studies, after adjusting for age and sex.

Variation in Inflammatory Biomarkers Among Demographic Groups Significantly Affects Their Accuracy in Diagnosing Periprosthetic Joint Infection.

BACKGROUND: Diagnosis of periprosthetic joint infection (PJI) is a multistep process that involves performing various tests including erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). The latter two tests, despite being used at all times for PJI diagnosis, are known to be nonspecific and substantially affected by demographic characteristics, including age, gender, race, and body mass index. It is unknown how these variations affect the diagnostic utility of serological markers for PJI. METHODS: Institutional databases were queried to identify patients undergoing revision arthroplasty between 2010 and 2018, in whom preoperative serum ESR and CRP was performed. Patient demographics were collected, and patients were cross-referenced with an internal database to determine their infection status. Analyses were performed to determine how ESR and CRP varied with respect to demographic factors, including age, gender, race, and infection status. Given that patient infection status was known at the time of revision, conclusions were drawn about the effect of these variations in inflammatory markers on the diagnostic utility of ESR and CRP. RESULTS: The value of ESR increased by age was higher in females and African American race. No significant differences were observed in the value of CRP among the demographic factors, although a slight positive trend was observed with respect to age. The variation in inflammatory markers significantly affected the sensitivity, specificity, and accuracy of ESR and CRP for PJI diagnosis. CONCLUSION: Understanding how the accuracy of diagnostic tests varies with respect to demographic factors can help physicians avoid subjecting patients to unnecessary additional testing and reach more accurate diagnoses of PJI.

Changes in Composition of ADAMTS-5 and CD-68 in the Knee Joint Synovial Fluid Cells of Meniscal Tears Patients an Immunohistochemical Study / Cambios en la Composición de ADAMTS-5 y CD-68 en las Células del Líquido Sinovial de la Articulación de la Rodilla en Pacientes con Desgarro de Menisco. Un Estudio Inmunohistoquímico

SUMMARY: Meniscus tear is an important injury affecting the quality of life. This work is aimed to investigate the activity of CD68 and ADAMTS-5 in cells in synovial fluid in male and female patients with meniscal tear. In this study ,18 male and 22 female patients with meniscal tears were included. Local pain sensation during patients' physical examination, swelling, performing daily activities and difficulty in running-walking complaints were determined. 5 cc synovial fluids were aspirated from the lateral suprapatellar pouch part of the knees with meniscal pain. After routine histological follow-up of the samples, they were embedded in paraffin and sectioned with microtome and 5 micrometer thickness. CD68 and ADAMTS-5 primary antibodies were used for immunohistochemical analysis. Sections were taken and evaluated with a stylish microscope. The distribution of blood cells after meniscus tear was higher in female patients than in male patients. CD68 distribution in female patients appeared higher than in male patients. CD68 expression was high in macrophage cell cytoplasm. ADAMTS-5 expression was higher in female patients in degenerative cells and apoptotic cells. ADAMTS-5 is an important metallo-protein involved in the development of apoptotic signal and extracellular matrix synthesis in patients with ADAMTS-5 meniscus tear, and it may be an important criterion for the treatment developed after injury. CD68 and ADAMTS-5 activity was thought to be one of the important signal pathways that can be identified in the treatment of meniscus tear.

RESUMEN: La rotura del menisco es una lesión importante que afecta la calidad de vida. El objetivo fue investigar la actividad de CD68 y ADAMTS-5 en células del líquido sinovial en pacientes masculinos y femeninos con desgarro meniscal. Se incluyeron 18 pacientes masculinos y 22 femeninos con desgarros meniscales. Se determinó la sensación de dolor local durante el examen físico de los pacientes, la hinchazón, la realización de actividades diarias y la dificultad al correr y caminar. Se aspiraron 5 cc de líquido sinoviale de la parte de la bolsa suprapatelar lateral de las rodillas de los pacientes con dolor meniscal. Después del seguimiento histológico de rutina, las muestras se incluyeron en parafina y se seccionaron con un micrótomo de grosor de 5 micrómetros. Para el análisis inmunohistoquímico se usaron los anticuerpos primarios CD68 y ADAMTS-5. La distribución de las células sanguíneas después del desgarro del menisco fue mayor en pacientes femeninos que en pacientes masculinos. La distribución de CD68 en pacientes femeninos fue más alta que en pacientes masculinos. Además la expresión de CD68 fue alta en el citoplasma de los macrófagos. La expresión de ADAMTS-5 fue mayor en pacientes femeninos en las células degenerativas y células apoptóticas. ADAMTS-5 es una metaloproteína importante en el desarrollo de la señal apoptótica y la síntesis de matriz extracelular en pacientes con rotura de menisco ADAMTS-5, y puede ser un criterio importante para el tratamiento después de la lesión. La actividad de CD68 y ADAMTS-5 era una de las vías de señal importantes que se pueden identificar en el tratamiento de la rotura del menisco.

Thirty days after anterior cruciate ligament transection is sufficient to induce signs of knee osteoarthritis in rats: pain, functional impairment, and synovial inflammation.

OBJECTIVE: To compare the unilateral signs of knee osteoarthritis (KOA) 30 and 60 days after anterior cruciate ligament transection (ACLT). Pain, gait function, synovial fluid inflammation, and histopathological changes in the synovial membrane were analyzed, as well as the interaction between the variables. MATERIALS AND METHODS: Male Wistar rats (n = 32; 219.2 ± 18.6 g) were randomly distributed into four groups of eight animals each. Two groups were submitted to unilateral ACLT surgery to induce KOA and analyzed after 30 (KOA30) and 60 days (KOA60). Two control groups (without surgery) were also assessed after the same time periods (C30 and C60). All the groups were evaluated before ACLT from the least to most stressful tests (skin temperature, mechanical response threshold, gait test, thermal response threshold, and joint swelling), as well as 30 and 60 days after surgery. After euthanasia, the synovial fluid and synovial membrane were collected. RESULTS: Thirty days after ACLT, KOA30 showed decrease paw print area and mechanical response threshold, higher joint swelling, skin temperature, leukocyte count, cytokine levels, and synovitis score. No differences were found between KOA30 and KOA60. CONCLUSION: Our data showed that 30 days after ACLT is sufficient to induce signs of KOA in rats, such as pain, functional impairment, and synovial inflammation, suggesting that a shorter time period can be used as an experimental model.

A Novel Adjunct Indicator of Periprosthetic Joint Infection: Platelet Count and Mean Platelet Volume.

BACKGROUND: Diagnosing a periprosthetic joint infection (PJI) is difficult and often requires a combination of objective findings. The purpose of this study is to determine whether platelets, a known acute phase reactant, would be able to further aid in the diagnosis of PJI. METHODS: A single-institution retrospective review study was performed on all revision total hip and knee arthroplasties done between 2000 and 2016 (n = 4939). PJI was defined by Musculoskeletal Infection Society criteria (n = 949). Platelet count and mean platelet volume were assessed from each patient's preoperative complete blood count. These values were then assessed as a ratio via receiver operating characteristic (ROC) curve analysis. RESULTS: The platelet count to mean platelet volume ratio for PJI patients was 33.45 compared to 25.68 for patients in the aseptic revision cohort (P < .001). ROC curve analysis demonstrates that a ratio of 31.70 has a sensitivity of 48.10 (95% confidence interval 44.9-51.4) and a specificity of 80.85 (95% confidence interval 79.6-82.1). This specificity was higher than that of both estimated sedimentation ratio (ESR) and C-reactive protein (CRP) for the same cohorts using optimal values determined via ROC curve analysis. When used in conjunction with ESR and CRP, there was a statistically significant increase in the diagnostic performance of the model used to assess PJI relative to the model that just employed ESR and CRP (P < .05). CONCLUSION: Our study demonstrates that platelets and their associated serum biomarkers are associated with PJI and warrant consideration in patients who are being evaluated for potential PJI.

Determination of diagnostic value (validity) leukocyte esterase (urine dipstick strip) in differentiating inflammatory arthritis from bacterial arthritis

Abstract Background: The current diagnostic cornerstone for septic arthritis contains gram stains, bacterial culture, and cell count with a differential of aspirated synovial fluid. Recently, a synovial leukocyte esterase (LE) test has been used for diagnosing septic arthritis. Since this test measures the esterase activity of leukocytes, there is always a dilemma for using this test in patients with inflammatory arthritis. Methods: We collected the synovial fluid specimens as part of the general diagnostic protocol for patients suspected of Juvenile Idiopathic Arthritis (JIA) or Septic Arthritis (SA). Each group included 34 patients. We compared the result of the synovial LE test with the result of the culture of each patient. Results: The mean ages of patients were 64.14 ± 31.27 and 50.88 ± 23.19 months in the JIA group and septic arthritis group, respectively. The LE test results were positive in 30 specimens, trace in 3 and negative in one in the first-time test and were positive in 31 specimens and trace in 3 in the second-time test, while it was negative in all patients with JIA. Hence, the sensitivity of the synovial LE test was 80.8%, the specificity, PPV, and NPV were 78.6, 70.0, 86.8% respectively based on a positive culture. Conclusion: The leukocyte esterase strip test can be used as a rapid, bedside method for diagnosing or excluding bacterial infections in different body fluids. The synovial LE test can be used as an accurate test to rapidly rule in or out an acute articular bacterial infection, even in patients with concurrent inflammatory arthritis.(AU)

Pro-inflammatory mediators and neutrophils are increased in synovial fluid from heifers with acute ruminal acidosis.

BACKGROUND: Acute ruminal acidosis (ARA) is a metabolic disease of cattle characterized by an aseptic synovitis. ARA is the result of an increased intake of highly fermentable carbohydrates that frequently occurs in dairy cattle subjected to high production requirements. In human joint diseases such as rheumatoid arthritis and gout, several pro-inflammatory molecules are increased in the synovial fluid, including cytokines, prostaglandin E2 (PGE2), metalloproteinases, and neutrophil extracellular traps (NETs). The aim of this study was to identify the presence of proinflammatory mediators and neutrophils in the synovial fluid of heifers with ARA, induced by an oligofructose overload. Five heifers were challenged with an oligofructose overload (13 g/kg BW) dissolved in water. As a control, a similar vehicle volume was used in four heifers. Synovial fluid samples were collected from the tarso-crural joint and PGE2, IL-6, IL-1ß, ATP, lactate dehydrogenase (LDH), albumin, glucose, matrix metalloproteinase-9 (MMP-9), cellular free DNA, NETs, and serpin B1 were analyzed at 0, 9, and 24 h post treatment. RESULTS: At 9 h post oligofructose overload, an increase of IL-1ß, IL-6, PGE2, serpin B1 and LDH was detected in the joints when compared to the control group. At 24 h, the synovial fluid was yellowish, viscous, turbid, and contained abundant neutrophils. An increase of DNA-backbone-like traps, histone 3 (H3cit), aggregated neutrophil extracellular traps (aggNETs), and serpin B1 were observed 24 h post treatment. Furthermore, albumins, LDH, ATP, MMP-9, IL-6, and IL-1ß were increased after 24 h. CONCLUSIONS: The overall results indicate that IL-1ß, IL-6 and PGE2, were the earliest proinflammatory parameters that increased in the synovial fluid of animals with ARA. Furthermore, the most sever inflammatory response in the joint was observed after 24 h and could be associated with a massive presence of neutrophils and release of aggNETs.

The differentiating ability of four plasma biomarkers in canine hip dysplasia.

BACKGROUND: The accumulation of cartilage breakdown products in body fluids has been extensively investigated to assess the accuracy of molecular biomarkers from a diagnostic, prognostic, and therapeutic perspective. Nevertheless, to the authors' knowledge, there is a lack of information about spontaneous models of hip osteoarthritis and the differentiating ability of collagen, noncollagen, and inflammatory biomarkers. OBJECTIVES: We aimed to assess the accuracy of four plasma biomarkers that could differentiate between healthy dogs and dogs with hip dysplasia. METHODS: Twenty-four dogs were used in this institutionally approved study (12 in the mild to severe hip dysplasia group; 12 in the control group). Plasma concentrations of biomarkers were compared. The ability of each marker to differentiate control from diseased dogs was assessed using an independent t-test, logistic regression, and receiving operating characteristics (ROC) analysis. RESULTS: Three biomarkers were significantly different between the two groups. The collagen marker procollagen type II propeptide (PIICP) was useful in differentiating between control and diseased dogs with the best combination of sensitivity and specificity. The four biomarkers showed high area under the curve (AUC) values. CONCLUSIONS: The results indicate that plasma biomarkers can be used as a screening tool for canine hip dysplasia. Although the cutoff values and diagnostic ability of the biomarkers used in this study show promising results, the sources of individual variability should be addressed. Future studies with larger groups of dogs are needed to correlate plasma levels in serum and synovial fluid during clinical disease.

In vitro and in vivo characterization of an interleukin-15 antagonist peptide by metabolic stability, 99m Tc-labeling, and biological activity assays.

Interleukin (IL)-15 is an inflammatory cytokine that constitutes a validated therapeutic target in some immunopathologies, including rheumatoid arthritis (RA). Previously, we identified an IL-15 antagonist peptide named [K6T]P8, with potential therapeutic application in RA. In the current work, the metabolic stability of this peptide in synovial fluids from RA patients was studied. Moreover, [K6T]P8 peptide was labeled with 99m Tc to investigate its stability in human plasma and its biodistribution pattern in healthy rats. The biological activity of [K6T]P8 peptide and its dimer was evaluated in CTLL-2 cells, using 3 different additives to improve the solubility of these peptides. The half-life of [K6T]P8 in human synovial fluid was 5.88 ± 1.73 minutes, and the major chemical modifications included peptide dimerization, cysteinylation, and methionine oxidation. Radiolabeling of [K6T]P8 with 99m Tc showed a yield of approximately 99.8%. The 99m Tc-labeled peptide was stable in a 30-fold molar excess of cysteine and in human plasma, displaying a low affinity to plasma proteins. Preliminary biodistribution studies in healthy Wistar rats suggested a slow elimination of the peptide through the renal and hepatic pathways. Although citric acid, sucrose, and Tween 80 enhanced the solubility of [K6T]P8 peptide and its dimer, only the sucrose did not interfere with the in vitro proliferation assay used to assess their biological activity. The results here presented, reinforce nonclinical characterization of the [K6T]P8 peptide, a potential agent for the treatment of RA and other diseases associated with IL-15 overexpression.

The 2018 Definition of Periprosthetic Hip and Knee Infection: An Evidence-Based and Validated Criteria.

BACKGROUND: The introduction of the Musculoskeletal Infection Society (MSIS) criteria for periprosthetic joint infection (PJI) in 2011 resulted in improvements in diagnostic confidence and research collaboration. The emergence of new diagnostic tests and the lessons we have learned from the past 7 years using the MSIS definition, prompted us to develop an evidence-based and validated updated version of the criteria. METHODS: This multi-institutional study of patients undergoing revision total joint arthroplasty was conducted at 3 academic centers. For the development of the new diagnostic criteria, PJI and aseptic patient cohorts were stringently defined: PJI cases were defined using only major criteria from the MSIS definition (n = 684) and aseptic cases underwent one-stage revision for a noninfective indication and did not fail within 2 years (n = 820). Serum C-reactive protein (CRP), D-dimer, erythrocyte sedimentation rate were investigated, as well as synovial white blood cell count, polymorphonuclear percentage, leukocyte esterase, alpha-defensin, and synovial CRP. Intraoperative findings included frozen section, presence of purulence, and isolation of a pathogen by culture. A stepwise approach using random forest analysis and multivariate regression was used to generate relative weights for each diagnostic marker. Preoperative and intraoperative definitions were created based on beta coefficients. The new definition was then validated on an external cohort of 222 patients with PJI who subsequently failed with reinfection and 200 aseptic patients. The performance of the new criteria was compared to the established MSIS and the prior International Consensus Meeting definitions. RESULTS: Two positive cultures or the presence of a sinus tract were considered as major criteria and diagnostic of PJI. The calculated weights of an elevated serum CRP (>1 mg/dL), D-dimer (>860 ng/mL), and erythrocyte sedimentation rate (>30 mm/h) were 2, 2, and 1 points, respectively. Furthermore, elevated synovial fluid white blood cell count (>3000 cells/µL), alpha-defensin (signal-to-cutoff ratio >1), leukocyte esterase (++), polymorphonuclear percentage (>80%), and synovial CRP (>6.9 mg/L) received 3, 3, 3, 2, and 1 points, respectively. Patients with an aggregate score of greater than or equal to 6 were considered infected, while a score between 2 and 5 required the inclusion of intraoperative findings for confirming or refuting the diagnosis. Intraoperative findings of positive histology, purulence, and single positive culture were assigned 3, 3, and 2 points, respectively. Combined with the preoperative score, a total of greater than or equal to 6 was considered infected, a score between 4 and 5 was inconclusive, and a score of 3 or less was not infected. The new criteria demonstrated a higher sensitivity of 97.7% compared to the MSIS (79.3%) and International Consensus Meeting definition (86.9%), with a similar specificity of 99.5%. CONCLUSION: This study offers an evidence-based definition for diagnosing hip and knee PJI, which has shown excellent performance on formal external validation.

The Alpha-Defensin Test for Diagnosing Periprosthetic Joint Infection in the Setting of an Adverse Local Tissue Reaction Secondary to a Failed Metal-on-Metal Bearing or Corrosion at the Head-Neck Junction.

BACKGROUND: In patients with adverse local tissue reaction (ALTR) secondary to a failed metal-on-metal (MoM) bearing or corrosion at the head-neck junction in a metal-on-polyethylene bearing, ruling in or out periprosthetic joint infection (PJI) can be challenging. Alpha-defensin has emerged as an accurate test for PJI. The purpose of this multicenter, retrospective study was to evaluate the accuracy of the alpha-defensin synovial fluid test in detecting PJI in patients with ALTR. METHODS: We reviewed medical records of 26 patients from 3 centers with ALTR that had an alpha-defensin test performed. Patients were assessed for PJI using the Musculoskeletal Infection Society criteria. Thirteen of these subjects had MoM total hip arthroplasty, 9 had ALTR secondary to head-neck corrosion, and 4 had MoM hip resurfacing. RESULTS: Only 1 of the 26 patients met Musculoskeletal Infection Society criteria for infection. However, 9 hips were alpha-defensin positive, including 1 true positive and 8 that were falsely positive (31%). All 8 of the false positives were also Synovasure positive, although 5 of 8 had an accompanying warning stating the results may be falsely positive due to a low synovial C-reactive protein value. CONCLUSION: Similar to synovial fluid white blood cell count, alpha-defensin testing is prone to false-positive results in the setting of ALTR. Therefore, we recommend an aggressive approach to ruling out PJI including routine aspiration of all hips with ALTR before revision surgery to integrate the synovial fluid blood cell count, differential, cultures and adjunctive tests like alpha-defensin to allow for accurate diagnosis preoperatively.

Analysis of synovial fluid of the Capybara's stifle joints.

BACKGROUND: Although normal synovial fluid has been well characterized in domestic animals such as dogs, cats, horses, and cows, the available information on larger rodents is scarce. OBJECTIVES: The purpose of the study was to analyze the physical, chemical, and cytologic characteristics of the synovial fluid in stifle joints of Capybaras. METHODS: Five free-ranging adult female Capybaras (Hydrochoerus hydrochaeris), weighing from 37 to 56 kg were used. Synovial fluid was obtained by aspiration of 10 stifle joints. Samples were analyzed for physical, chemical, and cytologic properties. RESULTS: Spontaneous clotting was negative in 9 samples. Most synovial fluids had pH 8, and protein concentrations ranged from 1.6 to 3.6 g/dL. The mucin clot test was good in all 6 samples that were tested. Nucleated cell counts ranged from 140 to 508 cells/µL. Relative differential leukocyte counts demonstrated a predominance of mononuclear cells (97.6%), including 76.2% undifferentiated mononuclear cells, 18.1% macrophages, and 3.66% lymphocytes. Polymorphonuclear cells included 1.83% neutrophils and 0.2% eosinophils. CONCLUSION: The synovial stifle joint fluid of healthy free-ranging adult Capybaras is clear, colorless, viscous, and with chemical features and cytologic findings similar to those seen in domestic animals.

Esterasa leucocitaria como prueba diagnóstica ante un proceso infeccioso articular de rodilla / Leukocyte esterase as a diagnostic tool for an infectious disease of the knee

Resumen: Antecedentes: La infección articular es un reto ortopédico por su complejidad diagnóstica y efectos devastadores al no tratarse oportunamente. Contamos con diversos estudios de diagnóstico: cultivo, VSG, PCR, conteo de leucocitos, entre otros, pero ninguno es preciso, tardan más de 30 minutos en realizarse y requieren de infraestructura compleja. En este estudio determinamos la sensibilidad y especificidad de la esterasa leucocitaria para la detección de un proceso infeccioso articular en población mexicana. Material y métodos: Durante Noviembre de 2015 a Abril de 2016, se obtuvo líquido sinovial de pacientes con diagnóstico de infección articular con o sin implante, y de otros sin infección, con patología degenerativa de rodilla. Se evaluó la muestra mediante el test de esterasa leucocitaria COMBI-SCREEN 11SYS con lectura colorimétrica a los dos minutos; se determinó positivo para infección con dos cruces; el resto de la muestra fue enviado a cultivo. Resultados: Realizamos el test en 64 muestras de líquido sinovial de rodilla, 19 diagnosticadas con infección articular y 45 sin infección. Se obtuvo una sensibilidad de 100%, especificidad de 88.24%, VPP de 68.42% y VPN de 100%; índice de Kappa de .753. Conclusiones: La esterasa leucocitaria es una prueba eficaz para detectar un proceso infeccioso contra uno inflamatorio con alta probabilidad de acierto. Este estudio presentó un índice de concordancia Kappa de 0.753, con lo que demostró ser reproducible, por lo que recomendamos implementarlo en los servicios de urgencias a nivel nacional.

Abstract: Background: Articular infection is an orthopedic challenge due to its difficult diagnosis and devastating results. Various diagnostic studies exist: culture, ESR, CRP, count of leukocytes, among others, but none is specific, they all take more than 30 minutes to complete, and require complex infrastructure. In this study, we determine the sensitivity and specificity of the leukocyte esterase for detection of an infectious process joint in Mexican population. Material and methods: During November 2015 to April 2016, we obtained synovial fluid from two groups of patients: one with a diagnosis of synovial joint infection with or without implant, and the control group, without infection but with degenerative pathology of the knee. We evaluated the sample using the leukocyte esterase test COMBI-SCREEN 11SYS with colorimetric reading at two minutes; two crosses determined positive for infection; the remainder of the sample was sent for culture. Results: We performed the test in 64 samples of synovial fluid, 19 diagnosed with articular infection and 45 without it. We obtained a sensitivity of 100%, specificity of 88.24%, PPV of 68.42%, and NPV of 100%; Kappa index of .753. Conclusions: Leukocyte esterase is an effective test to detect an infectious process against an inflammatory one with a high probability of success. This study presented an index of agreement Kappa of 0.753, proving to be reproducible.

[Leukocyte esterase as a diagnostic tool for an infectious disease of the knee]. / Esterasa leucocitaria como prueba diagnóstica ante un proceso infeccioso articular de rodilla.

BACKGROUND: Articular infection is an orthopedic challenge due to its difficult diagnosis and devastating results. Various diagnostic studies exist: culture, ESR, CRP, count of leukocytes, among others, but none is specific, they all take more than 30 minutes to complete, and require complex infrastructure. In this study, we determine the sensitivity and specificity of the leukocyte esterase for detection of an infectious process joint in Mexican population. MATERIAL AND METHODS: During November 2015 to April 2016, we obtained synovial fluid from two groups of patients: one with a diagnosis of synovial joint infection with or without implant, and the control group, without infection but with degenerative pathology of the knee. We evaluated the sample using the leukocyte esterase test COMBI-SCREEN 11SYS with colorimetric reading at two minutes; two crosses determined positive for infection; the remainder of the sample was sent for culture. RESULTS: We performed the test in 64 samples of synovial fluid, 19 diagnosed with articular infection and 45 without it. We obtained a sensitivity of 100%, specificity of 88.24%, PPV of 68.42%, and NPV of 100%; Kappa index of .753. CONCLUSIONS: Leukocyte esterase is an effective test to detect an infectious process against an inflammatory one with a high probability of success. This study presented an index of agreement Kappa of 0.753, proving to be reproducible.

ANTECEDENTES: La infección articular es un reto ortopédico por su complejidad diagnóstica y efectos devastadores al no tratarse oportunamente. Contamos con diversos estudios de diagnóstico: cultivo, VSG, PCR, conteo de leucocitos, entre otros, pero ninguno es preciso, tardan más de 30 minutos en realizarse y requieren de infraestructura compleja. En este estudio determinamos la sensibilidad y especificidad de la esterasa leucocitaria para la detección de un proceso infeccioso articular en población mexicana. MATERIAL Y MÉTODOS: Durante Noviembre de 2015 a Abril de 2016, se obtuvo líquido sinovial de pacientes con diagnóstico de infección articular con o sin implante, y de otros sin infección, con patología degenerativa de rodilla. Se evaluó la muestra mediante el test de esterasa leucocitaria COMBI-SCREEN 11SYS con lectura colorimétrica a los dos minutos; se determinó positivo para infección con dos cruces; el resto de la muestra fue enviado a cultivo. RESULTADOS: Realizamos el test en 64 muestras de líquido sinovial de rodilla, 19 diagnosticadas con infección articular y 45 sin infección. Se obtuvo una sensibilidad de 100%, especificidad de 88.24%, VPP de 68.42% y VPN de 100%; índice de Kappa de .753. CONCLUSIONES: La esterasa leucocitaria es una prueba eficaz para detectar un proceso infeccioso contra uno inflamatorio con alta probabilidad de acierto. Este estudio presentó un índice de concordancia Kappa de 0.753, con lo que demostró ser reproducible, por lo que recomendamos implementarlo en los servicios de urgencias a nivel nacional.