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PubMed2002

Phenotypic characterization of inflammatory cells from osteoarthritic synovium and synovial fluids.

Osteoarthritis (OA) is considered a degenerative joint disorder caused by mechanical wear to the articular surface. However, while joint injury, obesity, and mutations in collagen increase the risk of developing OA, evidence implicates inflammatory mechanisms in disease progression and chronicity. To address this question we used FACS analysis, immunohistochemistry, and in vitro cell culture to evaluate inflammatory mechanisms in synovial fluids and joint tissues obtained after arthrocentesis or knee replacement surgery. Immunohistochemistry revealed a significant T cell infiltrate in six of nine tissue specimens. T cells were present throughout the synovial membrane and were particularly localized around vasculature and in large cellular aggregates. Cells within the aggregates expressed markers associated with immune activation and antigen presentation. T cells from OA synovial fluids expressed an activated phenotype and synthesized interferon-gamma following in vitro stimulation. These data support the hypothesis that inflammatory cells play a significant role in OA disease progression and chronicity.

Aged↗
PubMed1990

Primary inflammatory reaction in synovial fluid and tissue in rabbit immobilization osteoarthritis.

The kinetics and composition of the primary cellular inflammatory process were studied in the synovial fluid (SF) and synovial tissue (ST) compartments of a rabbit knee immobilization osteoarthritis model. Immobilization induced rapid migration of neutrophils (59% +/- 26% of all cells) into SF in three days, which was accompanied by nonspecific esterase-positive monocytes (71% +/- 8% of all mononuclear cells). This finding suggests that non-specific inflammation mediated by phagocytic leukocytes predominates the cellular response in the SF compartment. In contrast, morphometric analysis of ST proper showed an inflammatory mononuclear cell response, the intensity of which diminished over time during the study period from Day 3 (416 +/- 59 cells per 0.049 mm2 ST tissue) through Day 10 (305 +/- 32 cells) to Day 35 (174 +/- 36 cells). A dotlike T-pattern alpha-naphthyl acetate esterase (ANAE) was found in the T-cell-dependent areas of secondary lymphatic tissue in the spleen, enabling immunocytologic ANAE marker studies. The ST response in situ was predominated by tissue macrophage, though infiltrates rich in T lymphocytes were present in the immediate sublining stroma. There was a significant correlation between the intensity of the SF cell response (total recovery) and the percentage of neutrophils, but there was no correlation between the intensity of the ST response and the proportion of T lymphocytes. These T-cell accumulations together with the local proliferation of fibroblastlike lining cells and stromal fibroblasts suggest that the primary inflammatory cell response is not caused by either wear and tear or mechanically by cartilage fragments.

Animals↗
PubMed1993

The structure of aggrecan fragments in human synovial fluid. Evidence that aggrecanase mediates cartilage degradation in inflammatory joint disease, joint injury, and osteoarthritis.

OBJECTIVE: To determine the proteolytic fragmentation patterns and N-terminal sequence of aggrecan fragments in human synovial fluid from patients with inflammatory arthritides, joint injury, or osteoarthritis (OA). METHODS: Knee synovial fluid was obtained from patients with joint injury, OA, acute pyrophosphate arthritis (pseudogout), reactive arthritis, psoriatic arthritis, or juvenile rheumatoid arthritis. Chondroitin sulfate-substituted aggrecan fragments present in the fluid were purified by cesium chloride gradient centrifugation and enzymatically deglycosylated. Core protein species were determined by N-terminal analysis and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with electroblotting and detection with monoclonal antibody 3B3. RESULTS: Samples from patients with joint injury, OA, and inflammatory joint disease all showed a similar 3-band pattern, with core sizes of approximately 200 kd, 170 kd, and 135 kd. In all samples, diffuse immunoreactive products were also seen, with an apparent size of > 250 kd. N-terminal analysis of core preparations of all samples showed a consistent single predominant sequence, beginning at alanine 374 of the human aggrecan core protein. CONCLUSION: The aggrecan fragments present in joint fluids from patients with various inflammatory arthritides, joint injury, or OA result from a predominant cleavage of the human aggrecan core protein at the glutamate 373-alanine 374 bond within the interglobular domain, between the G1 and G2 domains. The consistent pattern of fragments seen on SDS-PAGE and the single predominant N-terminal sequence suggest a common degradative mechanism of aggrecan in these different joint conditions. The identity of the proteolytic agent (aggrecanase), however, remains unknown. These results appear to have important implications with regard to the development of therapies to protect cartilage from degradation in patients with joint disease.

Adolescent↗
PubMed2002

Synovial fluid levels of E-selectin and intercellular adhesion molecule-1: relationship to joint inflammation in children with chronic arthritis.

E-selectin and intercellular adhesion molecule (ICAM)-1 are crucial to the inflammatory response in chronic inflammatory arthritis. Soluble (s) levels of these molecules in sera and synovial fluid (SF) correlate with some clinical parameters and synovial tissue expression of the same molecules in rheumatoid arthritis. Studies of sera from children with chronic inflammatory arthritis corroborate this information; corresponding SF data are relatively lacking. We thus studied SF sE-selectin and sICAM-1 in 28 children with active juvenile rheumatoid arthritis or a spondyloarthropathy. Levels were correlated with erythrocyte sedimentation rate (ESR), SF leukocyte counts, duration of disease, and duration of response to concomitant intra-articular corticosteroid injection. Levels were compared according to use of methotrexate and/or sulfasalazine. Synovial fluid sE-selectin correlated with ESR and SF leukocyte counts. There was a trend toward lower sICAM-1 in patients treated with sulfasalazine and/or methotrexate. We conclude that SF levels of sE-selectin accurately reflect intra-synovial inflammation. Soluble ICAM-1 levels may reflect the effects of disease-modifying agents.

Adolescent↗
PubMed1987

Carboxypeptidase N (kininase I) activity in blood and synovial fluid from patients with arthritis.

Carboxypeptidase N (CPN, kininase I) and kininase II (angiotensin converting enzyme) activities were measured simultaneously in blood plasma and synovial fluid in patients suffering from rheumatoid arthritis (RA), psoriatic arthritis (PA) and osteoarthritis (OA) and in the plasma of normal volunteers. CPN levels (defined as the rate of hydrolysis of furylacryloyl-Ala-Lys) in blood were modestly increased and correlated with erythrocyte sedimentation rate in RA and PA. Based on the hydrolysis of synthetic substrates, CPN activity was much higher than kininase II activity in synovial fluid (SF). SF kininase activities were always inferior to the blood levels in all patients and were correlated with the logarithm of SF leukocyte counts, an indicator of the intensity of inflammation. In addition, CPN and albumin levels in SF were highly correlated when expressed as a percent of the plasma concentrations. Biochemical properties of CPN in crude SF confirmed its similarity to blood CPN. Polymorphonuclear leukocytes derived from inflammatory SF did not release CPN. It is concluded that kininases diffuse from the blood into SF through increased vascular permeability and that CPN could be a major metabolic pathway for kinins in this form of exudate. CPN leads to the formation of des-Arg kinins, selective agonists of the B1 receptors for kinins.

Arthritis↗
PubMed1990

Salmonella-reactive synovial fluid T-cell clones in a patient with post-infectious Salmonella arthritis.

From a patient with reactive arthritis following Salmonella typhimurium enteritis, synovial fluid T-lymphocytes were cloned and expanded in vitro. Seven out of 74 clones showed a marked proliferative response to antigens of heat-killed Salmonella typhimurium with autologous T-cell-depleted peripheral blood mononuclear cells as antigen-presenting cells. The Salmonella-reactive clones were of the CD4+ phenotype, antigen-induced proliferation could be inhibited by a monoclonal antibody to HLA class II. One clone recognized both Salmonella and Campylobacter jejuni antigens in the proliferation assay. The multiclonality of Salmonella-reactive synovial fluid T-cells indicates that the microorganisms have been present, at least transiently, within the affected joint and thus recruited specific T-lymphocytes that might initiate synovitis.

Adult↗
PubMed1975

[Rheumatoid synovial fluid].

In making a comparative study of different parameters in the examination of synovial fluid (rheumatoid factors, ragocytes, leucocytes, synovial complement, immunoglobulins) the authors show that rheumatoid fluid has characteristics that make differentiation from other inflammatory arthropathies possible on a statistical basis, thanks to the test for the homogeneity of the means. The presence of these characteristics seems to constitute a diagnostic argument which can be measured by mathematical methods.

Arthritis↗
PubMedDate not supplied

Concentrations of prostaglandins D2, E2, F2 alpha, 6-keto-F1 alpha and thromboxane B2 in synovial fluid from patients with inflammatory joint disorders and osteoarthritis.

The concentrations of PGD2, PGE2, PGF2 alpha, 6-keto-PGF1 alpha and TXB2 in synovial fluid from patients with rheumatoid arthritis (RA), Reiter's disease (RD), acute gouty arthritis (GA) and osteoarthritis (OA) were measured by radioimmunoassay. PGE2 was found to be the most predominant prostanoid (pg/ml; Mean +/- S.E.M.): RA 887 +/- 85, RD 870 +/- 71, GA 1064 +/- 155 and OA 665 +/- 71. In patients with OA lower mean levels of all the prostanoids were found than compared to the other groups of patients. Only in patients with RA a slight correlation between PGD2/PGF2 alpha, PGE2/PGF2 alpha and PGE2/6-keto-PGF1 alpha could be demonstrated. No significant correlations between the leucocyte cell counts in the synovial fluid and the prostanoid concentrations were found. In patients with RA developing recurrent knee joint effusions within four weeks after the first sampling significantly lower levels of PGE2 and TXB2 were found in the recurrent samples (PGE2 792 +/- 183; TXB2 179 +/- 33) than compared with the original samples (PGE2 984 +/- 146; TXB2 239 +/- 32).

Arthritis↗
PubMed1985

The effect of synovial fluid proteins in the degradation of hyaluronic acid induced by ascorbic acid.

The degradation of hyaluronic acid induced by ascorbic acid and the effect of synovial fluid proteins, such as ceruloplasmin, transferrin, and albumin, were investigated on the basis of the elution volume and the molecular weight of hyaluronic acid using high-performance gel permeation chromatography. Hyaluronic acid was degraded to less than one-third of the original molecular weight in the range of the physiological concentrations of ascorbic acid. Synovial fluid proteins protected against the ascorbate-dependent degradation of hyaluronic acid at their physiological concentrations. It is suggested that the inhibitory activity of ceruloplasmin mainly depends on the ferroxidase activity and that of transferrin is probably due to iron binding property.

Albumins↗
PubMed1991

Activated fibrinolytic enzymes in the synovial fluid during acute arthritis induced by urate crystal injection in dogs.

To identify the relationship of the severity of inflammation and fibrinolytic activity in arthritis, the fibrinolytic activity of synovial fluid was studied in acute experimental arthritis induced by injecting monosodium urate crystals into dogs' knee joints. The maximum activity in the synovial fluid was observed 6 h after crystal injection. It was inferred that the fibrinolytic activity was mainly due to plasminogen activator based on fibrin plate assays, substrate specificity, inhibitor effects and zymography. On the other hand, the activity of lysosomal enzymes (beta-glucuronidase and cathepsin G) reached a peak in the synovia after 12 h. Histological examination of the synovial membrane after 12 h also showed greater inflammation than at 6 h. The peak in fibrinolytic activity preceded the peak of lysosomal enzymes and histological changes. These results suggest that an increase in fibrinolytic activity by plasminogen activator may contribute to the development of an acute inflammatory response.

Acute Disease↗
PubMed1983

The demonstration of an interleukin-2 like activity in the synovial fluids of rheumatoid arthritis patients.

It has been shown that synovial tissues and fluids contain activated T lymphocytes. We examined the synovial fluids of rheumatoid arthritis patients and control (noninflammatory) fluids for the presence of interleukin 2 (IL-2) activity, a lymphokine essential to maintaining cells in an activated state. Using an IL-2 dependent T cell line it was found that 10/14 (71%) of rheumatoid synovial fluids contained IL-2 like activity while 0/8 control fluids demonstrated activity. The active material was resistant to heat (56 degrees C for 1h), was less than 50,000 molecular weight by gel filtration fractionation and was not precipitated by 50% saturated ammonium sulphate. It is therefore not immune complex in nature.

Arthritis, Rheumatoid↗
PubMed1994

[Tumor necrosis factor in synovial fluids of temporomandibular joint dysfunction syndrome].

With the MTT tetrazolium WEHI 164 clone 13 cell cytotoxicity assay, we measured TNF alpha (tumor necrosis factor) activity in synovial fluids of TMJDS (Temporomandibular joint dysfunction syndrome). We found no detected TNF alpha level from 5 patients with muscle dysfunction, raised TNF alpha levels from 5 of 11 patients with internal derangement and from 9 of 11 patients with organic destruction (osteoarthritis). The findings of biologically active TNF alpha in synovial fluids of TMJDS suggest that TNF alpha may play a role in the pathogenesis of TMJDS.

Adolescent↗
PubMed1997

Biochemical and immunochemical evaluation of tissues and synovial fluid from patients undergoing total joint arthroplasty.

Cytokines are inflammatory mediators responsible for numerous clinical conditions, and are thought to lead to the resorption of bone. Understanding the nature of the cells producing these factors which control the resorption of bone will ultimately lead to a better understanding of why implants fail or integrate. In this study, synovial tissues and synovial fluids were processed for biochemical as well as histochemical and immunohistochemical determination cytokines responsible for bone resorption. The results from this study showed by both quantitative enzyme linked immunoassay (ELISA) and qualitatively by immunohistology a marked increase (twofold) in interleukin-1 (IL-1), and tumor necrosis factor-beta (TNF beta) in synovial tissues in comparison to control tissues of cartilage, ligament and meniscus. Evaluation of tissues both immunochemically and by Hematoxylin and Eosin demonstrated the presence of fibroblast and cells such as macrophages, and multinucleated giant cells in the synovium that are capable of producing bone resorption. Synovial fluid from primary and revision patients were evaluated for TNF beta and IL-1 were not statistically different. Overall, the results indicate that the inflammatory cells of the synovium are secreting factors which may act to mediate aseptic loosening of implants.

Arthroplasty, Replacement, Knee↗
PubMed1996

The clinical significance of cytoplasmic inclusions(CPI) in synovial fluid examination.

The clinical significance of cytoplasmic inclusions(CPI) in synovial fluid(SF) examination was evaluated. We examined SF specimens collected from major rheumatology clinics in the Philadelphia area during the period of January to December 1995. Among 759 patients in the initial study group, 419 cases with established diagnoses and full synovial analyses were included. Their diagnoses and SF analysis results including leukocyte counts, differential counts and wet preparations were collected and analysed. Ninety seven of the 419 SF specimens were found to have CPI. CPI were found in SF from almost all rheumatic diseases. They were most likely to be found in inflammatory arthropathy including rheumatoid arthritis(RA, 46%), juvenile rheumatoid arthritis(JRA, 78%) and psoriatic arthritis(55%). On the contrary, CPI were least common in crystal-induced arthropathy among the inflammatory arthropathy. CPI were found 8 out of 98 gout cases(8%) and 2 among 53 calcium pyrophosphate dihydrate(CPPD) deposition disease(4%). In noninflammatory arthropathy, CPI were found in only 6 cases(6%) out of the 103 osteoarthritis(OA). In RA cases with non-inflammatory SF, 4 of the 20 SF(20%) had CPI while only 6% of OA SF had CPI. OA SF with CPI were all noninflammatory SF. In summary, CPI were a common finding on SF examination. CPI were more likely to be found in inflammatory arthropathy than noninflammatory. Among inflammatory arthropathy, CPI can favor non-crystal arthropathy than crystal arthropathy. Awareness of the presence of CPI is suggested as an addendum to routine SF analysis. Renewed investigation of the several types of CPI may add further to the understanding of joint disease.

Inclusion Bodies↗
PubMed1988

Human parvovirus B19 DNA in synovial fluid.

We describe a 33-year-old woman with a serologically proven human parvovirus B19 infection, who developed synovitis. Using a dot-blot hybridization technique, we detected B19 DNA in her synovial fluid. To our knowledge, this is the first report of the isolation of B19 from synovial fluid.

Adult↗
PubMed1989

Sickled cells in synovial fluid: clue to unsuspected hemoglobinopathy.

In this report, we have described three patients who had hemarthroses, with sickled red blood cells discovered by analysis of synovial fluid. On the basis of this observation, each patient was evaluated for the presence of abnormal hemoglobins, and each was found to have a hemoglobinopathy that was previously unsuspected. These patients differ from those in other reports in that two of the three had no associated arthritic condition that could readily explain synovitis or a condition that predisposed them to bleeding into a joint. Although the accumulated evidence suggests that heterozygous hemoglobinopathies do not produce arthritic syndromes, these reports again raise that question. We cannot conclude, however, that the hemarthroses were definitively caused by the underlying hematologic abnormality. Important when synovial fluid is mixed with blood, since other medical conditions can be diagnosed if abnormal findings are detected.

Aged↗
PubMedDate not supplied

Mononuclear cells in human synovial fluid. Identification of lymphoblasts in rheumatoid arthritis.

A simple method was developed to identify large mononuclear (LMN) cells in human synovial fluid based on morphology and staining with Sudan black B. All cells were classified as monocyte-derived macrophages (MDM), lymphoblasts (LB), or synovial lining cells (SLC). Lymphoblasts were seen in 58 of 60 rheumatoid fluids (mean: 69 +/- 18% LB per LMN cells). However lymphoblasts were rarely seen in synovial fluids from patients with crystal-induced synovitis or bacterial infections.

Arthritis, Rheumatoid↗
PubMed1990

Interleukin 2 (IL 2) inhibitor in rheumatoid synovial fluid: correlation with prognosis and soluble IL 2 receptor levels.

A soluble activity inhibiting over 50% of the CTLL-2 cell line response to recombinant human interleukin 2 (IL 2) was found in 17 of 29 (59%) rheumatoid synovial fluids. To study the prognosis value of this activity, 16 rheumatoid synovial fluids were collected before a radiation synovectomy of the knee with 7 mCi of 90Yt. Patients with a good clinical result after the synovectomy had a lower IL 2 inhibitory activity than those with a bad or incomplete result (P less than 0.01). Levels of inhibitory activity and of soluble IL 2 receptors were correlated with each other and with the response of the synovitis to the radiation synovectomy. These results extend the clinical usefulness of soluble IL 2 receptor measurements and indicate a correlation between the immune activation of the rheumatoid synovitis and its clinical activity.

Adult↗