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PubMed1975

Hydroxyproline levels and collagenolytic activity in synovial fluids of patients with rheumatic diseases.

Free, total, and peptide hydroxyproline levels were determined in synovial fluid obtained from the knee joints of 60 patients with theumatoid arthritis (RA), and 26 patients with degenerative joint disease. In addition, in 160 synovial fluid samples obtained from 121 patients including 50 with degenerative joint disease, 60 with RA, 3 with Reiter's syndrome, 3 with hydarthrosis intermittens and 5 with ankylosing spondylitis, the collagenolytic activity was determined. The mean values of free and peptide hydroxyproline in the inflammatory and degenerative fluids were the same, but slight differences were found in the mean values of total hydroxyproline. No effect on the level of free and bound hydroxyproline was observed after treatment with intra-articular hydrocortisone and gold salts. The collagenolytic activity of synovial fluid was registered in 38% of cases of RA and in some cases of Reiter's syndrome and hydrarthrosis intermittens, but it was not found in 50 cases of degenerative joint disease or in cases of ankylosing spondylitis. During a longer observation of patients with inflammatory forms of RA a variability in the collagenolytic activity was observed in repeated examinations of the fluid obtained from the same patient; this activity appeared and disappeared. The incidence of collagenolytic activity and its values were higher in patients with active rheumatoid process and this activity was present more frequently in patients with a short history of the disease (up to 3 years). The collagenolytic activity of rheumatoid fluids was, to a high degree, inhibited by normal human serum. The problem of presence or lack of collagenolytic activity in rheumatoid fluids is discussed.

Adult↗
PubMed1989

Cells of the monocyte-macrophage series in peripheral blood and synovial fluid in inflammatory arthritis. A preliminary study of cellular phenotype.

The phenotype of peripheral blood and synovial fluid cells of the monocyte-macrophage series was studied by FACS to extend previous immunohistological observations made in synovial membranes. The results of this preliminary study suggest that changes can be found in peripheral blood monocytes which might influence diapedesis and interaction with immune complexes. The phenotype of synovial fluid macrophages does not provide evidence of advanced maturation and is most consistent with rapid recruitment of the cells into the joint. Synovial fluid macrophages showed increased expression of the leukocyte integrin p150.95 which functions as an adhesion molecule and complement receptor. This is consistent with our previous observation of upregulation of this molecule on Mph in inflamed synovial membranes. This preliminary study has indicated areas for more detailed analysis with the aim of defining sequential changes in macrophage expression of functional surface molecules in inflammatory arthritis.

Adult↗
PubMed1995

Effect of tumor necrosis factor antibody on synovial fluid cytokine activities in equine antebrachiocarpal joints injected with endotoxin.

Six horses received intra-articular injections of a mixture of 1 micrograms of endotoxin/5 mg of equine tumor necrosis factor (eqTNF) monoclonal antibody in 1 antebrachiocarpal joint and an equal volume (2 ml) of 1 micrograms of endotoxin/5 mg of control antibody in the opposite joint. Synovial fluid sample collection (1 ml) was accomplished by use of an indwelling, intra-articular catheter at postinjection hours (PIH) 0, 1, 1.5, 2, 5, and 8, and by arthrocentesis at PIH 24. Joint fluid samples were analyzed for nucleated cell count, protein concentration, and TNF, interleukin 6 (IL-6), IL-1, and IL-1-inhibitory activities. To monitor local inflammation, each carpus was graded semiquantitatively for swelling prior to each sample collection. Tumor necrosis factor, IL-1, or IL-1-inhibitory activity was not detected in any synovial fluid sample collected before endotoxin/antibody was administered. However, low IL-6 activity (< 100 U/ml) was found in 2 of 12 preinjection samples. In joints injected with endotoxin/control antibody mixture, maximal mean +/- SEM activities for TNF (1,019 +/- 310 U/ml), IL-1 (173 +/- 102 U/ml), and IL-6 (10.8 +/- 3.1 x 10(4) U/ml) were observed at PIH 2, 5, and 8, respectively. Tumor necrosis factor and IL-1 activities returned to baseline values by PIH 8 and 24, respectively; however, IL-6 activity remained high. Interleukin 1 inhibitory activity (27.4 +/- 2.25 IU/ml) was detected in all PIH-24 samples from control joints, but was not detected at any other time in control joints (limit of detection, 20 IU/ml). Tumor necrosis factor activity was not detected in any synovial fluid sample from joints treated with endotoxin/eqTNF antibody. In contrast, endotoxin IL-1 inhibitory activity (PIH 24) was higher in eqTNF antibody-treated joints (41.0 +/- 7.7 IU/ml) than in control joints, but the difference was not significant. Mean WBC count and protein concentration in control and treated joints were maximal at PIH 8. The curves for mean values of WBC count and total protein concentration were not significantly different in treated versus control joints. Swelling in each treated joint was either less than or the same as that in the opposite control joint at even, time in the initial 8 PIH. There was significant (P = 0.043) difference between treated and control joints at PIH 5 and 8. These results describe a profile of synovial fluid TNF, IL-1, IL-6 bioactivities, and IL-1-inhibitory activity during the initial 24 hours of synovitis induced by intra-articular administration of endotoxin in horses. Our eqTNF monoclonal antibody was effective in neutralizing TNF activity in synovial fluid when administered intra-articularly with endotoxin in horses. The induction of IL-1, IL-1 inhibitory activity IL-6, WBC, and total protein concentration responses are largely independent of TNF activity in synovial fluid of horses receiving endotoxin intra-articularly.

Animals↗
PubMed1993

Patterns of T-cell receptor variable beta gene expression by synovial fluid and peripheral blood T-cells in rheumatoid arthritis.

Conflicting data have been reported regarding the presence or absence of a predominant variable (V) region T-cell receptor (TCR) gene in the peripheral blood or synovial fluid of patients with rheumatoid arthritis (RA). In this study we have used the polymerase chain reaction (PCR) to compare the level of TCR V beta gene expression by peripheral blood mononuclear cells (PBMC) and by synovial fluid cells obtained from HLA DRB1 *0401 and *0404 RA patients. PCR was performed using cDNA synthesized from freshly obtained cells (not stimulated in vitro). The pattern of expression observed for most of the V beta genes studied showed either preferential expression by PBMC or similar levels of expression between PBMC and synovial fluid T-cells. However, among individual patients (N = 5), several V beta genes were identified that were expressed to a significantly greater degree by synovial fluid cells. V beta 14 expression was detected in PBMC of all patients and the level of transcripts encoding V beta 14 increased following stimulation with immobilized anti-CD3 and IL-2. In vitro manipulation of populations of T-cells was found to alter the level of expressed V beta gene products. A V beta gene common to all patients that was consistently deleted from PBMC or expressed to a greater degree by resident, unsorted synovial fluid cells compared to PBMC was not identified.

Adult↗
PubMed2005

Examination of synovial fluid and serum following intravenous injections of hyaluronan for the treatment of osteoarthritis in dogs.

A randomized, blinded, prospective clinical trial was performed to determine the effects of intravenous (i.v.) administration of hyaluronan sodium (HA) on serum glycosaminoglycans (GAG) concentrations, synovial fluid (SF) hyaluronan concentrations and viscosity in dogs treated for unilateral rupture of the cranial cruciate ligament. Twenty-two dogs undergoing tibial plateau leveling osteotomy were used in this study. Synovial fluid from both stifles and serum were collected prior to surgery and at 2, 4, and 8 weeks following surgery. Dogs received either 1.0 ml (10 mg) of sodium hyaluronate (treatment group 1; n = 10) or equal volume of 0.9% NaCl (treatment group 2; n = 12), i.v. immediately, 2 and 4 weeks following surgery. Synovial fluid viscosity was evaluated using a magnetically driven, acoustically tracked, translating-ball rheometer. Synovial fluid HA disaccharide content was measured by fluorophore-assisted carbohydrate electrophoresis. Serum GAG concentrations were measured by alcian blue spectrophotometric assay. Data were analyzed using a Wilcoxon sign rank test (p < 0.05). Mean +/- SD viscosity (cP) was significantly higher (p = 0.011) in SF obtained from the intact stifle (450 +/- 604.1) than injured (54.8 +/- 60.8) prior to surgery. Mean +/- SD HA concentrations (ug/ml) were significantly higher (p = 0.02) in synovial fluid obtained from the injured stifles (281.4 +/- 145.9) than intact stifles (141.6 +/- 132.5). No significant difference was noted within or between treatment groups in SF viscosity, HA concentrations, or serum GAG concentrations at any time following surgery. Stifles with cranial cruciate ligament insufficiency had significant alterations in SF viscosity and HA concentrations.

Adjuvants, Immunologic↗
PubMed1989

Prognostic significance of synovial fluid analysis in rheumatoid arthritis.

Thirty patients with definite rheumatoid arthritis and hydropsy in a knee joint were followed for 42 months in a prospective study. The knee joints were initially aspirated and 15 synovial fluid variables investigated. The patients were split into two groups, those with and those without progress of radiologically detected destruction in the knee joints during the follow-up. Of the synovial fluid variables at the start synovial fluid proteins (P = 0.002) and acid phosphatase (P = 0.03) differed statistically significantly between the groups, both being higher in patients with worsening of knee joint. The results suggest that high synovial fluid proteins and acid phosphatase are predictors of poor prognosis in a joint affected by rheumatoid arthritis.

Adult↗
PubMedDate not supplied

[LE cells in the synovial fluid].

The occurence of LE cells in the synovial fluids of two patients is reported. The demonostration of LE cells in synovial exudate is clinically valuable. Examination of joint fluid should therefore pay attention to the search for LE cells and their precursors. Reoccurrence of E-cells in the synovial fluid indicates their origin in the joint cavit. They could still be found even after multiple intraarticular injections of triamcinolone acetonide.

Arthritis, Rheumatoid↗
PubMed1994

Absence of Mycobacterium tuberculosis DNA in synovial fluid from patients with rheumatoid arthritis.

OBJECTIVES: To determine whether Mycobacterium tuberculosis DNA can be detected in synovial fluid of patients with rheumatoid arthritis (RA). METHODS: The polymerase chain reaction was applied to cellular components of synovial fluid. RESULTS: No evidence of M tuberculosis DNA was found in synovial fluid from 31 patients with RA and 13 control patients. CONCLUSION: The findings do not support a role for persistent M tuberculosis infection in the pathogenesis of RA.

Adolescent↗
PubMed1991

Oxygen derived free radicals and synovial fluid hyaluronate.

High performance liquid chromatography with TSK 5000 PW or TSK 6000 PW size exclusion columns combined with a 125I labelled hyaluronic acid binding protein assay was used to study the effects of oxygen derived free radicals on synovial fluid hyaluronate. A continuous flux of free radicals was generated by the xanthine oxidase/hypoxanthine system. When the free radical flux was generated with xanthine oxidase/hypoxanthine in the presence of the iron chelator desferrioxamine and the hydroxyl radical scavenger mannitol a 30-50% decrease in hyaluronate peak was detected, but the molecular weight of synovial fluid hyaluronate remained almost unchanged as a result of reaction with superoxide radicals and hydrogen peroxide. When trace amounts of iron and EDTA were present in the reaction mixture depolymerisation of synovial fluid hyaluronate occurred, and it reached a final molecular weight of about 13,500 daltons. These results suggest that superoxide and hydroxyl radicals may have a different mode of action on synovial fluid hyaluronate. Superoxide radicals and hydrogen peroxide do not induce depolymerisation but, rather, change the molecular configuration of synovial fluid hyaluronate.

Carrier Proteins↗
PubMed2005

Analysis for crystals in synovial fluid: training of the analysts results in high consistency.

BACKGROUND: Identification of monosodium urate (MSU) and calcium pyrophosphate dehydrate (CPPD) crystals in synovial fluid samples is diagnostic of gout and CPPD crystal related arthropathy. Various studies have shown poor consistency in results of crystal analysis. OBJECTIVE: To determine whether training of the analysts increases the consistency. METHODS: An expert rheumatologist gave a course on crystal detection and identification. The four trained observers then blindly and independently examined synovial fluid samples previously classified by the expert which had been obtained from patients with both crystal arthropathies and other non-crystal related inflammatory joint conditions. RESULTS: 194 observations were made on 64 synovial fluid samples: 96 without crystals (49.4%), 55 with CPPD crystal (28.4%), and 43 with MSU crystals (22.2%). For crystal detection (presence or absence of crystals), sensitivity was 95.9% and specificity 86.5%. For identification of MSU crystals, sensitivity was 95.3% and specificity 97.2%. For identification of CPPD crystals, sensitivity was 92.7% and specificity 92.1%. The kappa index of agreement with the reference standard between the observers was 0.84 for any crystal detection, 0.93 for MSU crystal sample identification, and 0.79 for CPPD crystal sample identification. CONCLUSIONS: For trained observers, the detection and identification of crystals in synovial fluid is a consistent procedure.

Arthritis↗
PubMed2005

Intracellular and extracellular CPPD crystals are a regular feature in synovial fluid from uninflamed joints of patients with CPPD related arthropathy.

OBJECTIVES: To determine whether calcium pyrophosphate dihydrate (CPPD) crystals can be found in the synovial fluid of non-inflamed joints in patients with CPPD related arthropathy; if so, to determine whether they interact with cells and produce subclinical inflammation in this setting. METHODS: 74 synovial fluid samples were obtained from non-inflamed knees of 74 patients with CPPD related arthropathy. Identification of CPPD crystals and synovial fluid cell counts were done manually in undiluted samples using a haematocytometric chamber. A supravital stain (Testsimplets, Boehringer Mannheim) was used to carry out differential counts and to assess the presence of intracellular crystals. RESULTS: All 74 samples contained CPPD crystals. The mean cell count was 301.4 cells/microl (95% confidence interval (CI), 216.6 to 386.4; range 22 to 2302.5). Mononuclear cells accounted for 83.2% (95% CI, 80.4% to 86.1%; range 43% to 99%), the rest being polymorphonuclear (PMN) cells (16.8% (95% CI, 13.9% to 19.6%; range 1% to 57%)). All the samples contained intracellular CPPD crystals, which were found in 24.0% of all the cells (95% CI, 20.1% to 27.9%; range 1% to 78%). Most of the intracellular crystals were inside mononuclear cells (22.2% of all the cells (95% CI, CI 18.5% to 25.9%)), although some PMN also contained them (1.8% of all the cells (95% CI, 1.1% to 2.4%)). CONCLUSIONS: CPPD crystals are normally found in synovial fluid of non-inflamed joints of patients with CPPD related arthropathy, and they interact with cells. The raised cell counts and percentage of PMN suggest mild subclinical inflammation in these joints.

Adult↗
PubMed2003

Cytokine profile in serum and synovial fluid of arthritis patients with Chlamydia trachomatis infection.

Chlamydia trachomatis (Ct)-induced arthritis (CtIA) is characterized by persistent Ct infection, which stimulates secretion of cytokines in vitro. We therefore investigated whether CtIA patients have a unique cytokine profile in synovial fluid or serum in vivo. Because underlying Ct infection is overlooked in a high percentage of patients with initially diagnosed undifferentiated oligoarthritis (UOA), we examined whether determination of cytokines might also be of diagnostic relevance for this arthritis form. Matched serum and synovial fluid specimens from 26 patients with CtIA were analyzed and compared to those from 34 patients with UOA in whom Ct infection was excluded and those of nine patients with rheumatoid arthritis (RA). In 15 CtIA patients, Ct DNA from synovial fluid could be amplified by polymerase chain reaction. The following cytokine or cytokine antagonists were measured by enzyme-linked immunosorbent assay: interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, IL-6, IL-1 receptor antagonist, and soluble TNF receptor p75. No statistically significant differences in cytokine levels between patients with CtIA or the other arthritis forms were detected. Also, comparison between CtIA patients with (n = 17) and without Chlamydia DNA (n = 9) in synovial fluid revealed no significant differences for these cytokines. Cytokine levels in serum and synovial fluid were not different between CtIA, UOA without Ct infection, and RA patients. The intracellular presence of Ct was not associated with a specific profile of these cytokines in vivo.

Adult↗
PubMed1987

Synovial fluid analysis for diagnosis of amyloid arthropathy.

To detect the presence of amyloid, we studied synovial fluid samples from 7 patients who had biopsy-proven amyloidosis and clinical arthropathy and from 98 patients who had other forms of arthritis. Centrifuged sediments from synovial fluids of all 7 patients with amyloid and clinical arthropathy were positive on staining with Congo red, whereas the 98 others were negative. Under polarized microscopy, the amyloid appeared as an amorphous material with strong Congo red uptake and typical apple-green birefringence. These findings indicate that analysis of synovial fluid sediments with Congo red may represent a simple, yet sensitive, test for the diagnosis of amyloid arthropathy.

Aged↗
PubMedDate not supplied

[The physical properties of the synovial fluid as a lubrication of joints].

The properties of the synovial fluid and the results of the electrophysical studies of the fluid by the thermoelectret method are briefly described. The interrelation between current peaks on the thermostimulated current spectra of thermostimulated currents of the synovial fluid and the liguid-crystalline state of its components was established. The results can be used for developing the methods of diagnostics of synovial joint diseases and design of artificial joints.

Humans↗
PubMedDate not supplied

Abnormal T cell receptor V beta gene expression in the peripheral blood and synovial fluid of rheumatoid arthritis patients.

OBJECTIVE: The aim of this study was to assess T cell receptor V beta-gene expression in the peripheral blood and synovial fluid of rheumatoid arthritis patients. METHODS: Cytometric analysis was performed on peripheral blood and synovial fluid lymphocytes from 12 patients using a restricted set of V beta-specific monoclonal antibodies (to V beta 5.1-3, V beta 6.7 and V beta 8). In 5 patients the expression of the V beta 1 through V beta 20 gene families was also analysed, using a recently described method based on a one-side-specificity polymerase chain reaction coupled to reverse dot hybridization. RESULTS: Cytometric analysis failed to show any consistent difference in the expression of V beta 5, 6 and 8 between the two compartments on the one hand, or between the peripheral blood of normal individuals and patients on the other hand. The PCR/dot hybridization method did not demonstrate a significant difference in the V beta repertoires between peripheral blood and synovial fluid samples from arthritis patients. However, in all patients the V beta 6, 13 and/or 14 families were expressed to a high level, so that these families frequently represented over 40% of the V beta 1-20 repertoire in both compartments, instead of the approximately 20% seen in normal peripheral blood samples. CONCLUSION: We conclude that V beta 6, 13 and 14 are overexpressed in both the peripheral blood and synovial fluid of rheumatoid arthritis patients compared to normal samples.

Arthritis, Rheumatoid↗
PubMed2000

Matrix metalloproteinases 2 and 9 activity in bovine synovial fluids.

Matrix metalloproteinases (MMPs) are important enzymes found in connective tissues and thought to be involved in cartilage degradation. They are detectable in bovine synovial fluid and may play a destructive role in bovine septic arthritis. The MMP gelatinase enzymes were detected by gelatin zymography using image analysis of the gels. The active gelatinase levels were determined by a gelatin degradation enzyme-linked immunosorbent assay (ELISA). Increased concentrations of MMP-9 activity were found in the synovial fluids of cows with septic arthritis (P < 0.001) in comparison with fluids from normal joints. Using the gelatin degradation ELISA the net active gelatinases were measured, and significant increases were found in gelatinase bioactivities in synovial fluids from septic joint disease cases (P < 0.001). Increased concentrations of MMP-2 activity were found in the synovial fluids of cows with aseptic arthritis, which appeared to be playing an important role in degradation of articular cartilage in joint disease. This finding required further investigation.

Animals↗
PubMed2002

CD69 expression on lymphocytes and interleukin-15 levels in synovial fluids from different inflammatory arthropathies.

OBJECTIVE: The aim was to study a possible relationship between CD69 expression on lymphocytes and interleukin-15 (IL-15) levels in synovial fluid (SF) from patients with different inflammatory arthropathies. METHODS: CD69 expression was assessed by two-color flow cytometry on different subsets of synovial fluid lymphocytes (SFL) obtained from patients with diagnoses of rheumatoid arthritis (RA), seronegative spondyloarthropathy (SSd), and crystal-associated arthritis (CAA). The IL-15 levels in synovial fluid supernatants were measured by enzyme-linked immunoassay (ELISA). RESULTS: No significant differences between the three groups of arthropathies were observed in the distribution of synovial fluid lymphocyte subsets. CD69-positive SFL were mainly CD3-, CD45RO-, and CCR5-positive cells. Although no significant differences in the percentage of CD69-positive lymphocytes were observed between the three groups of patients, the highest level of CD69 expression on lymphocytes was observed in RA patients (mean fluorescence intensity 37.9 +/- 5.2 compared to 17.5 +/- 3.3 in SSd and 12.4 +/- 2.6 in CAA, P = 0.007 and P < 0.001, respectively). In addition, the expression of CD69 on SFL from RA correlated with their respective IL-15 levels measured in SF supernatants. CONCLUSION: Our data provide in vivo evidence of the putative role that IL-15 can play in the high expression of CD69 on SFL from RA patients.

Aged↗
PubMed2003

Elevated angiogenin levels in synovial fluid from patients with inflammatory arthritis and secretion of angiogenin by cultured synovial fibroblasts.

Angiogenesis is a key process in the pathogenesis of inflammatory arthritis. Angiogenin is one of the most potent inducers of neovascularization in experimental models in vivo. To look for evidence that angiogenin is involved in inflammatory joint disease, we examined plasma and synovial fluid (SF) samples from rheumatology patients and synovial fibroblast cell culture supernatants. Angiogenin levels were determined by radioimmunoassay and ELISA. Plasma angiogenin concentrations ranged from 96 to 478 ng/ml, with no significant difference between patients and normal controls. In SF, angiogenin concentrations were significantly higher in patients with acute or chronic synovitis (rheumatoid arthritis (RA): median, 104 ng/ml; range 13-748, n = 14; crystal-induced arthritis (CIA): median, 149 ng/ml; range, 37-616, n = 14, and other chronic inflammatory arthritis: median, 42 ng/ml; range, 15-205; n = 9) than in the 18 patients with osteoarthritis (OA) (median, 20 ng/ml; range 8-116) (P < 0.0001, anova). Angiogenin levels in SF from RA patients in remission with secondary OA were similar to those achieved in primary OA, and decreased in parallel with the resolution of acute gout. Angiogenin protein was released by cultured synovial fibroblasts from OA and RA patients, and reached 1.18 ng/106 cells/day. These data suggest that angiogenin may mediate local inflammation in arthritis via effects on angiogenesis and leucocyte regulation.

Analysis of Variance↗