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PubMed1982

Inflammatory activities of synovial fluids from rheumatoid- and osteo-arthritis to guinea pig skin.

For initial study to examine the chemical pathogenesis of rheumatoid and non-rheumatoid arthritis, the nature and inflammatory activities of 12 synovial fluids from rheumatoid arthritis and 10 from osteoarthritis were studied. Both rheumatoid and non-rheumatoid synovial fluids were found to have high protein concentration, acid and neutral protease activities. When injected 0.1 ml intradermally to normal guinea pig skin, both synovial fluids induced a strong vascular permeability at 0 minutes, mild one at 60 minutes, and mild inflammation which comprised histologically moderate leukocyte infiltration around the venules, swelling of venular endothelial cells, edema, and degeneration of collagen fibrils in the deep dermis of the guinea pig skin 4 hours following the injection. However, these parameters of inflammation induced by synovial fluids from both arthritis were of same degree in intensity. The identification of chemical mediators of inflammation in the synovial fluids of both arthritis requires further study.

Adult↗
PubMedDate not supplied

The influence of synovial fluid on lymphocyte functions in rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic recurrent and systemic inflammatory disease affecting around 1% of the population, that primarily involves the joints. In this study, we determined the Th1/Th2 lymphocytes ratio at the site of rheumatoid inflammation and the influence of the synovial fluid (SF) on the secretory and proliferative function in synovial fluid mononuclear cells (SFMC) and peripheral blood mononuclear cells (PBMC), obtained from patients with RA. Our results showed significant differences concerning the mononuclear cells and the CD4/CD8 ratio in synovial fluid and peripheral blood of patients. In SF prevailed Th1 cells, while in peripheral blood we found another cytokine profile of T lymphocytes. Also synovial fluid lymphocytes had a low PHA-stimulated blastogenic response. Patients plasma and synovial fluid showed an inhibitory effect on prolipheration indexes.

Arthritis, Rheumatoid↗
PubMedDate not supplied

Glycosaminoglycans in the synovial fluids of patients with juvenile rheumatoid arthritis.

A simple method was used to isolate hyaluronic acid (HA) and sulfated glycosaminoglycans from synovial fluids obtained from children with juvenile rheumatoid arthritis (JRA) and from normal age-matched controls. The bulk of the glycosaminoglycans present in both normal and pathologic synovial fluids consisted of hyaluronic acid and chondroitin 6-sulfate. The diseased synovial fluids showed a sharp decrease in the concentration of glycosaminoglycans when compared with normal controls. These findings are similar to those reported for the synovial fluid of adult patients with rheumatoid arthritis.

Arthritis, Juvenile↗
PubMed1981

DNA in synovial fluid and the circulation of patients with arthritis.

DNA levels were measured in synovial fluids and sera of 106 patients with rheumatoid arthritis (RA), osteoarthritis (OA), gout, pseudogout, and posttraumatic arthritis (TRA). In synovial fluids, the highest concentration was found in rheumatoid arthritis (mean +/- SE 18 +/- 3 microgram/ml for seropositive and 9 +/- 1 microgram/ml for seronegative variants), gout and pseudogout (17 +/- 3 microgram/ml). In contrast, the levels in patients with OA or acute TRA were very low: 0.8 +/- 0.1 microgram/ml an 1.1 +/- 0.2 microgram/ml, respectively. The differences between the means of the first disease group and OA or TRA is statistically significant. A similar pattern was observed for DNA levels in the circulation: in rheumatoid arthritis, the mean concentration was 135 +/- 28 ng/ml and 164 +/- 39 ng/ml for seropositive and seronegative RA, respectively. Again the levels in OA and TRA were much lower, 52 +/- 18 ng/ml and 0 ng/Ml, respectively. The latter are not significantly different from the mean levels of 95 normal, healthy controls (14 +/- 3 ng/ml), whereas the concentration of DNA in the serum of RA patients is significantly higher than in OA, TRA, or normal controls. Serial determinations of DNA and other criteria of disease activity (leukocytes and protein levels in synovial fluid, blood sedimentation rate) in individual patients revealed a strong correlation of elevated values with active episodes. THe results suggest that these parameters reflect tissue damage.

Arthritis↗
PubMed2006

Cytologic analysis of synovial fluid in clinically normal tarsal joints of young camels (Camelus dromedarius).

BACKGROUND: Camels are important in the racing industry and for milk, meat, and hair production in the Middle East. Evaluation of synovial fluid is an important part of the assessment of musculoskeletal injuries in this species. Information in the literature regarding synovial fluid in camels is limited. OBJECTIVES: The objective of this study was to determine the protein and cellular composition of synovial fluid from the tarsal joints of clinically normal, young camels (Camelus dromedarius). METHODS: Thirty clinically healthy, male camels, aged 9 to 12 months, were used in the study. Synovial fluid samples were collected from the right and left tarsal joints. Samples were processed within 60 minutes after collection. Total nucleated cell counts (TNCC) were assessed using a hemacytometer. Total protein concentration was determined using a refractometer. RESULTS: Forty-six samples were analyzed. The TNCC (mean +/- SD) was 175.8 +/- 136.7 cells/microL (range 50-678 cells/microL). Differential cell percentages were obtained for lymphocytes (58.2 +/- 21.55%, range 15-90%), monocyte/macrophages (38.3 +/- 20.8%, range 10-85%), and neutrophils (3.5 +/- 5.1%, range 0-15%). Protein concentration was 2.1 +/- 0.6 g/dL (range 1-3 g/dL). Significant differences were not observed in any parameters between right and left tarsal joints. CONCLUSION: Synovial fluid reference values were established and may be useful in the clinical investigation of joint disease in young camels.

Animals↗
PubMed1979

[Degradation products of fibrinogen in synovial fluid].

The fibrin/fibrinogen degradation products were measured in 24 synovial fluids sampled from patients with knee joint effusion. It has been observed that they are increased in synovial fluid involved in inflammatory processes. Their presence and concentration seem to be related to the control of local inflammation.

Arthritis↗
PubMed1987

Cartilage proteoglycans in synovial fluid and serum in patients with inflammatory joint disease. Relation to systemic treatment.

Proteoglycan concentrations in knee joint synovial fluid and in serum from patients with various inflammatory arthritides were studied using an enzyme-linked immunosorbent assay. Patients with reactive arthritis, calcium pyrophosphate arthropathy, and juvenile rheumatoid arthritis (age less than or equal to 20 years) had the highest synovial fluid concentrations. These values differed significantly (P less than 0.001) from those in patients with rheumatoid arthritis, psoriatic arthropathy, and chronic HLA-B27-associated arthropathy. Rheumatoid arthritis patients receiving low-dose prednisolone treatment had higher synovial fluid (P = 0.006) and serum (P less than 0.001) proteoglycan concentrations than did those taking nonsteroidal antiinflammatory drugs or slow-acting antirheumatic drugs. Serum proteoglycan concentrations were near the detection limits, and did not correlate with levels found in paired samples of knee joint synovial fluid. Patients with calcium pyrophosphate arthropathy had the highest mean serum level of proteoglycan. This assay of proteoglycan antigens is a useful tool in the study of proteoglycan metabolism in patients with joint disease. With its use, differences between disease groups and effects of therapy can be distinguished.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed1995

Concentration and size distribution of insulin-like growth factor-I in human normal and osteoarthritic synovial fluid and cartilage.

The concentration of free insulin-like growth Factor-I (IGF-I) and its complexes was determined in human normal and osteoarthritic synovial fluids, using ultrafiltration through 20- and 100-kDa membranes, followed by a radioimmunoassay of each fraction. In addition, freshly obtained samples of normal and osteoarthritic cartilage were incubated for several days, at both 4 and 37 degrees C. The incubation media (desorbates) were analyzed the same way as the synovial fluid samples to yield the concentration of IGF-I in cartilage in situ. Our findings are (i) Free IGF-I content is extremely low in both human serum and synovial fluid and there is no significant difference between the two; (ii) The concentration of total IGF-I in normal human synovial fluid is an order of magnitude lower than that in serum due mainly to the decrease in the concentration of the large complex; (iii) Preliminary results show that the total IGF-I in osteoarthritic synovial fluids is twice as high as in normal fluids; (iv) In normal human cartilage the levels of IGF-I in all its forms are very low and are consistent with the expected exclusion of large molecules by the extracellular matrix; (v) By contrast, in osteoarthritic cartilage, the concentrations of all forms of IGF-I are high, probably due to increased permeability of the matrix and binding; (vi) The levels of IGF-I found in normal human cartilage are more than an order of magnitude lower than those which stimulate proteoglycan synthesis in human cartilage in culture, while the IGF-I levels in osteoarthritic cartilage lie in the range in which stimulation does occur.

Adult↗
PubMed1966

Studies on the physiological chemistry of the tibio-tarsal synovial fluid of healthy bovines.

The physical properties, chemical composition and cytological content of the synovial fluid of healthy native breed cattle, obtained by tibio-tarsal arthrocentesis, was studied. Normally the synovial fluid of bovines is a clear, colourless or straw coloured, viscous, non-coagulable liquid with a normal mucin precipitate quality. The fluid is alkaline in reaction and with a lower specific gravity than serum. Bovine tibio-tarsal fluid exhibits no erythrocytes and presents a low leucocytic count which is dominated by lymphocytes and monocytes. Compared to bovine serum, synovial fluid contains almost the same levels of sugar, urea, creatinine and phosphorous; slightly lower calcium concentration and rather higher chlorides. The fluid is devoid of any traces of bilirubin. Young calves showed a significantly lower total leucocyte number, chloride level and sugar content and a higher calcium and inorganic phosphorous than bulls.

Animals↗
PubMed2001

Demonstration of mast cell chemotactic activity in synovial fluid from rheumatoid patients.

OBJECTIVES: The significance of the mast cell in the pathogenesis of rheumatic diseases has become more evident. Although mast cell hyperplasia is a feature of rheumatoid arthritis, the nature of mast cell chemoattractants involved in the recruitment of mast cells in joint diseases has not been studied in any detail. In this study the presence of mast cell chemotactic activity in synovial fluids was examined. METHODS: Synovial fluids from seven rheumatoid patients were tested in a modified Boyden chamber, where a human mast cell line was used as responder. The presence of stem cell factor (SCF) and transforming growth factor beta (TGFbeta) was measured by enzyme linked immunosorbent assay (ELISA). RESULTS: Six of the seven synovial fluids tested exhibited mast cell chemotactic activity. Two characterised human mast cell chemotaxins, SCF and TGFbeta, were highly expressed in the synovium. Soluble SCF could be detected in all fluids analysed. Blocking antibodies against SCF or TGFbeta almost completely blocked the activity in one fluid, partially blocked the activity in three, and did not affect the activity in two. Treatment of the responder cells with pertussis toxin reduced the migratory response against seven fluids, indicating the presence of chemoattractants mediating their effect through G(i) coupled receptors. CONCLUSION: These data demonstrate the presence of multiple factors in synovial fluid acting as mast cell chemoattractants, two of which are SCF and TGFbeta that contribute to the effect. These findings may be of importance for developing new strategies to inhibit mast cell accumulation in rheumatic diseases.

Adult↗
PubMed2006

Synovial fluid macrophages are capable of osteoclast formation and resorption.

To determine whether synovial fluid (SF) macrophages isolated from the SF of osteoarthritis (OA), rheumatoid arthritis (RA) and pyrophosphate arthropathy (PPA) joints are capable of osteoclast formation, and to investigate the cellular and humoral factors required for this to occur, SF macrophages (CD14+) were isolated from the knee joint SF from patients with OA, RA and PPA and cultured for up to 14 days with macrophage-colony stimulating factor (M-CSF) and soluble receptor activator for nuclear factor-kappaB ligand (RANKL) or tumour-necrosis factor-alpha (TNFalpha) and interleukin-1alpha (IL-1alpha). Osteoclast differentiation was assessed by expression of tartrate-resistant acid phosphatase (TRAP) and vitronectin receptor (VNR), F-actin ring formation and lacunar resorption. Osteoclast formation and lacunar resorption was seen in RANKL-treated cultures of SF macrophages isolated from OA, RA and PPA joints with the largest amount of resorption noted in RA and PPA SF macrophage cultures. In TNFalpha/IL-1alpha-treated RA and PPA SF macrophage cultures, osteoclasts capable of lacunar resorption were also formed. Lacunar resorption was more extensive in RANKL than TNFalpha/IL-1alpha-treated cultures. These findings indicate that SF macrophages are capable of differentiating into mature osteoclasts capable of lacunar resorption. M-CSF in combination with RANKL or TNFalpha/IL-1alpha was required for osteoclast formation. As inflammatory synovial fluids contain an increase in the number of macrophages and an increase in the amounts of RANKL, TNFalpha and IL-1alpha, these findings suggest that one means whereby bone erosions may form in rheumatoid or crystal arthritis is by differentiation of synovial fluid macrophages into osteoclasts.

Aged↗
PubMed2002

Urokinase, a constitutive component of the inflamed synovial fluid, induces arthritis.

Urokinase plasminogen activator (uPA) is an important regulator of fibrinolysis in synovial fluid. An increase of uPA activity and expression of its receptor have been reported in joints of patients with rheumatoid arthritis (RA). The aim of the present study was to assess the arthritogenic capacity of uPA and the mechanisms by which this effect is mediated. uPA was injected into the knee joints of healthy mice, and morphological signs of arthritis were assessed 4 days after the injection. The prerequisite of different leukocyte populations for the development of uPA-triggered arthritis was assessed by selective cell depletion. The inflammatory capacity of uPA was assessed in vitro. Finally, levels of uPA were measured in 67 paired blood and synovial fluid samples from RA patients. The synovial fluid from RA patients displayed higher levels of uPA compared with blood samples. Morphological signs of arthritis were found in 72% of uPA-injected joints compared with in only 18% of joints injected with PBS (P < 0.05). Synovitis was characterised by infiltration of CD4-Mac-1+ mononuclear cells, by the formation of pannus and by occasional cartilage destruction. The absence of monocytes and lymphocytes diminished the frequency of synovitis (P < 0.01), indicating an arthritogenic role of both these leukocyte populations. Synthetic uPA inhibitor downregulated the incidence of uPA-triggered arthritis by 50%. uPA induced arthritis, stimulating the release of proinflammatory cytokines IL-6, IL-1beta and tumour necrosis factor alpha. Accumulation of uPA locally in the joint cavity is a typical finding in erosive RA. uPA exerts potent arthritogenic properties and thus may be viewed as one of the essential mediators of joint inflammation.

Adult↗
PubMed2003

CD44H and the isoforms CD44v5 and CD44v6 in the synovial fluid of the osteoarthritic human knee joint.

OBJECTIVE: To determine whether the concentrations of CD44H and v5 and v6 in the synovial fluid are correlated with the presence of accompanying synovitis in the osteoarthritic joint and with the grade of osteoarthritis. DESIGN: Using antero-posterior and lateral X-rays of the knee joint and patellar view of 46 patients were graded with the Kellgren & Lawrence scale. Synovial fluid from these patients with different grades of primary osteoarthritis of the knee joint with and without synovial inflammation (synovitis) was collected during surgical procedures. A horseradish peroxidase conjugated anti-CD44H-, anti-sCD44v5- or anti-sCD44v6-antibody was added and labeled with tetramethylbenzidine. The absorbance was measured at wavelengths of 450/620 nm. Regression analysis was performed and the statistical significance was assessed, using the Student t-test for unequal variance. RESULTS: CD44H and v5 and v6 were detected in the synovial fluid of all 46 patients. Osteoarthritic patients with synovial inflammation showed significantly higher levels of CD44H and v6, but not v5, than osteoarthritic patients without synovial inflammation. With progression of osteoarthritis from Kellgren grade II to III, CD44v5 decreased significantly. All other isoform concentrations remained statistically unchanged. CONCLUSIONS: CD44H and the isoforms v5 and v6 were present in the synovial fluid of osteoarthritic patients. Their concentrations do not reflect the osteoarthritic stage in the Kellgren grading scale. CD44H and CD44v6, but not CD44v5, are significantly up-regulated in osteoarthritic synovial inflammation.

Adult↗
PubMed1992

The B cell repertoire in rheumatoid arthritis. III. Preferential homing of rheumatoid factor-producing B cell precursors in the synovial fluid.

OBJECTIVE: We sought to compare the frequencies of precursors producing IgM rheumatoid factors (IgM-RFs) in synovial fluid and peripheral blood B cells from patients with rheumatoid arthritis (RA). METHODS: We used limiting-dilution analysis of Epstein-Barr virus-activated B cells from seropositive and seronegative patients. B cell precursors producing IgM against insulin, an irrelevant autoantigen, were also assessed for comparison. RESULTS: On average, IgM-RF-producing precursors were 15-fold higher in the synovial fluid than in the peripheral blood of seropositive RA patients, but not in seronegative RA patients. The frequency of B cell precursors producing IgM against insulin was lower in the synovial fluid than in the peripheral blood of both the seropositive and the seronegative patient groups; moreover, the concentrations were similar in both groups. CONCLUSION: The findings provide evidence against a nonspecific accumulation of IgM-producing cells in the synovial fluid, and suggest that there is an active attraction of the RF-producing B cell precursors toward sites of inflammation in RA.

Adult↗
PubMed1986

C1 activation, with C1q in excess of functional C1 in synovial fluid from patients with rheumatoid arthritis.

Free Clq, in functionally active form was present in increased amounts in the synovial fluid of patients with rheumatoid arthritis. The presence of free Clq was associated with low concentrations of hemolytic C1, low C4 and raised amounts of C3dg/d fragments in the synovial fluid. The findings suggested intra-articular C1 activation with dissociation of C1 into free C1q and complexes containing C1r, C1s, and C1 inactivator. However, the immunochemical properties of synovial fluid C1r-C1s-C1 inactivator complexes appeared to differ from those of the complexes formed in serum, which hampered quantification with the assay used. Control patients with osteoarthritis or spondylarthritic syndromes did not show evidence of intra-articular complement activation, even though 1 patient with Reiter's disease had unexplained low concentrations of synovial fluid C4 and C3. The concentrations of circulating complement components were largely normal in the patients. Slightly increased concentrations of free C1q and C1r-C1s-C1 inactivator complexes in serum and C3dg/d fragments in EDTA plasma were observed, particularly in the patients with rheumatoid arthritis.

Adult↗
PubMed1999

[Detection of Borrelia DNA in synovial fluid for diagnosis of Lyme arthritis].

AIM: To test sensitivity and specificity of a polymerase chain reaction (PCR) targeting the Borrelia specific outer surface protein (Osp) A gene in synovial fluid for the diagnosis of Lyme arthritis, and thus permit an earlier start to treatment. PATIENTS AND METHODS: Prospectively we examined the synovial fluid of 37 patients with the clinical diagnosis of Lyme arthritis or with other arthropathies of known or unknown origin, searching for the presence of detectable borrelial DNA in both arms of the study. Retrospectively we examined the stored synovial fluid from 50 patients of the Department of Rheumatology of the University Hospital, Berne, with the clinical diagnosis of monarthritis or oligoarthritis of unknown etiology, juvenile chronic arthritis or rheumatoid arthritis. The laboratory biologist was unaware of the clinical diagnosis. RESULTS: In the prospective study no true false positive results were found: of the 28 patients without strong clinical suspicion of Lyme arthritis 27 were PCR negative. In one case with positive PCR for borrelial DNA the diagnosis could not be clarified, Lyme arthritis remaining a possibility. Therefore the specificity in the prospective study was at least 96%. Borrelial DNA in the synovial fluid was found in 5 out of 9 patients with strong clinical suspicion of Lyme arthritis. All 7 patients in this group were new, untreated cases. All the 5 PCR positive results belonged to this group, thus the "sensitivity" of the tested method was 71% in untreated cases of Lyme arthritis. In the retrospective study we found borrelial DNA in the synovial fluid of 2 patients. These 2 patients had gonarthritis of unknown origin. Retrospectively these 2 cases could be diagnosed as Lyme arthritis. CONCLUSION: In cases with clinical suspicion of Lyme arthritis the PCR method targeting a borrelial Osp A gene fragment common to all 3 European genospecies shows very good specificity and in untreated cases acceptable sensitivity. Introduction of the method studied into clinical practice is justified.

Adolescent↗
PubMed1986

Influence of C3 level on the determination of C3d in plasma and synovial fluid by radial immunodiffusion.

The influence of C3 levels on the determination of C3d in plasma and synovial fluid by radial immunodiffusion was investigated. In the method used, C3 is precipitated by 11% polyethylene glycol (PEG), and C3d is measured in the supernatant. In 51 healthy donors, a weak though significant correlation between C3 and C3d levels was found. The mean concentration of C3d was 1.6% of that in aged serum from healthy donors. So, small amounts of C3 (i.e., 1-2% of the normal plasma level) in the 11% PEG supernatants may contribute significantly to the C3d levels measured. A radioimmunoassay that detects C3, C3b, iC3b and C3c was used to measure C3 levels in the PEG supernatants. In PEG supernatants of 4 plasma samples, 0.3-0.6% of the C3 level in normal plasma was found, whereas in those of 2 synovial fluids much higher levels were found (4-10% of the normal plasma level). When purified 125I-labeled antibodies against C3c were added to the gel of the radial immunodiffusion, C3c antigen was detected in the precipitation rings obtained with all PEG supernatants of plasma samples from patients. Therefore, the quantitative contribution of C3 to the precipitation rings in the C3d radial immunodiffusion was analyzed after the addition of an excess of anti-C3c antibodies to the gel. No effect on the size of the C3d-precipitation rings obtained with plasma samples from patients was observed. However, the C3d precipitation rings obtained with synovial fluids were significantly smaller when the gel used in the radial immunodiffusion contained an excess of anti-C3c antibodies together with the anti-C3d serum. We conclude that it is necessary to add an excess of anti-C3c antibodies to the gel used for the radial immunodiffusion, for the determination of C3d levels in synovial fluid. An antiserum against human C3b, which contains both anti-C3c and anti-C3d antibodies, can be used for this purpose.

Blood Donors↗
PubMed1989

Characterisation of soluble and cell associated phospholipase A2 from rheumatoid synovial fluid.

The hydrolysis of radiolabelled Escherichia coli phospholipids, and micellar dispersions of phosphatidylethanolamine and phosphatidylcholine, were used to characterise the phospholipase A2 activity in synovial fluid from patients with rheumatoid arthritis. Cell free fractions of synovial fluid contain a phospholipase A2 enzyme that preferentially releases [14C]oleic acid from E coli biomembranes (specific activity 291.3 (SEM 27.6) pmol/min/mg). This enzyme requires calcium and is optimally active at neutral pH. Purified dispersions of phosphatidylethanolamine are also readily degraded by the soluble enzyme, but it is not active against phosphatidylcholine. The substitution of [14C]oleic acid by [3H]arachidonic acid for the labelling of E coli allowed differentiation between the soluble phospholipase A2 and the cell associated phospholipase A2 present in sonicates of mononuclear cells and neutrophils from peripheral blood and synovial fluid. The cell associated phospholipase A2 preferentially releases [3H]arachidonic acid from E coli cardiolipin. In this paper the phospholipid substrate specificity of phospholipase A2 from rheumatoid synovial fluid, the optimal assay conditions for its detection, and a standardised expression of activity in terms of pmol per minute per mg of protein are established.

Arthritis, Rheumatoid↗