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PubMed1984

Morphologic, quantitative and cytoenzymologic studies of synoviocytic and monocytic cells in synovial fluid.

The findings are presented of a morphologic, quantitative, cytochemical and cytoenzymologic study of the mononucleated nonlymphoid cells in knee synovial fluids from osteoarthritis and various inflammatory diseases. The morphologic criteria allowed the identification of subtypes, including phagocytic subtypes, among synoviocytic and monocytic cells in the fluids. The quantitative study showed an important afflux of monocytes and a hyperexfoliation of synoviocytes in the inflammatory diseases. In fluids with intermediate cellularity, the ratio of monocytes to synoviocytes allowed the differential cytodiagnosis between osteoarthrosis and arthritis. All monocytic subtypes, especially the phagocytic one, were highly significantly increased in the inflammatory diseases. A lower increase was shown by the synoviocytic subtypes, except the phagocytic one, which was not changed. Giant multinucleated synoviocytes were found in every type of disease and thus do not constitute a cytodiagnostic marker. Alcian blue staining without hyaluronidase treatment showed hyaluronate in only a small percentage of the synoviocytes. Cytoenzymologic study showed that synoviocytes and monocytes were positive for all tested hydrolases (beta glucuronidase, acid phosphatase and alpha naphthyl acetate esterase), with the reactivities always higher in the synoviocytes. The synoviocytes were always negative with peroxidase, so this reaction, although it marks only a minority of the monocytic population, can be used as an extra cytologic criterion for the discrimination of mononucleated cells in synovial fluid. There was no significant quantitative difference at the cellular level between osteoarthrosis and arthritides in the reaction to these four enzymes. The lysosomal enzymatic activity in both monocytic and synoviocytic cells confirmed their heterophagic properties. However, synoviocytic heterophagy seems to be a physiologic process, either little or not affected by inflammatory events. On the other hand, monocytic heterophagy and then the macrophagic transformation of monocytes appears to be a major aspect of intrasynovial inflammatory reactions. The question remains as to why, if a large majority of exfoliated synoviocytes comes from type A synovial-lining cells and if they belong to mononuclear phagocytic system, do they so weakly, or not at all, participate as phagocytes in the inflammatory reaction.

Acid Phosphatase↗
PubMed1992

Pyrophosphate in synovial fluid and urine and its relationship to urinary risk factors for stone disease.

Inorganic pyrophosphate (PPi) measurement in urine and synovial fluid has been established using the PPi-dependent phosphorylation of fructose-6-phosphate and subsequent reduction of dihydroxyacetone phosphate by NADH. The assay is linear up to 200 mumol/L, easy to perform and gives results comparable to more complex methods. Daily urinary output of PPi was independently related to both age (P = 0.0014) and sex (P = 0.0002). Men had higher values than women and older individuals excreted greater amounts. Male stone formers, younger than 45 years, had lower values than age matched male controls (P = 0.012). Younger female stone formers also tended to have lower values. In stone formers' urine significant and independent correlations were found of PPi excretion with urine volume (P = 0.004) and with phosphate excretion (P = 0.008). Oxalate excretion and that of other urine constituents and the degree of supersaturation with common stone-forming salts were not correlated with PPi. PPi excretion was markedly elevated in the urine of two patients with hypophosphatasia. The PPi concentration in synovial fluid from painful, swollen knee joints was elevated, but unrelated to the presence or absence of PPi or urate crystals.

Adolescent↗
PubMed1996

Plasminogen activators and their inhibitors in synovial fluids from normal, osteoarthritis, and rheumatoid arthritis knees.

OBJECTIVES: To establish baseline concentrations of plasminogen activators and their inhibitors in normal knee synovial fluids, and to compare them with well characterised osteoarthritis (OA) and rheumatoid arthritis (RA) knee fluids. METHODS: A total of 26 normal subjects, 71 patients with OA, and 17 patients with RA underwent knee aspiration. Patients with OA were subclassified according to presence of nodal generalised OA (NGOA) and synovial fluid calcium pyrophosphate crystals. Clinical assessment of inflammation (graded 0-6) was undertaken in OA and RA patients. Plasminogen activator (PA), plasminogen activator inhibitor (PAI), and urokinase-type PA receptor (uPAR) antigen concentrations were determined by enzyme linked immunosorbent assay. The species of PAs present were determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. RESULTS: Concentrations of all antigens (uPA, tissue-type PA (tPA), uPAR, and PAI-1), were significantly greater in RA than OA; those in OA were significantly greater than normal. The concentrations showed no direct association with clinically assessed inflammation of the knee. In normal fluids, no associations with age were observed. Antigen concentrations (uPA, tPA, and uPAR) in NGOA differed from those in other subclasses of OA, but the species of PA present did not appear to vary between disease groups. The predominant PA appeared to have identity with uPA. CONCLUSION: Because of the greater concentrations of these antigens in OA compared with normal fluids, OA cannot be used as a surrogate normal control in studies of the PA/PAI system. Alteration of the PA/PAI system was confirmed in RA and OA knee fluids, with greater changes evident in RA. The finding of different concentrations of PA antigens in NGOA compared with other OA fluids further supports a different pathogenic mechanism in this subset.

Adult↗
PubMed2002

Aberrant responsiveness to RANTES in synovial fluid T cells from patients with rheumatoid arthritis.

OBJECTIVE: To study expression and function of the chemokine receptor CCR5 in synovial fluid (SF) T cells from patients with rheumatoid arthritis (RA). METHODS: Expression of CCR5 was studied by flow cytometry and immunoblotting. The chemotactic response of T cells to chemokines was studied in cell migration assay. Tyrosine phosphorylation of Crk-associated substrate lymphocyte-type (CasL) was evaluated in immunoprecipitation and immunoblotting. RESULTS: SF T cells showed an increase in the population of CCR5, CXCR4, and CD45RO positive cells and exhibited an increase in chemotactic activity, which was not augmented with RANTES but stromal cell-derived factor-1alpha. Tyrosine phosphorylation per CasL molecule was markedly enhanced in SF T cells. In H9 cells, tyrosine phosphorylation of not only focal adhesion kinase but also CasL was induced after treatment with RANTES. Downmodulation of CCR5 by RANTES was decreased and recycling of CCR5 was accelerated in SF T cells when compared with peripheral blood (PB) T cells. When CD45RO positive PB T cells were cultured with interleukin 2, blunted responsiveness to RANTES-induced chemotaxis was reproduced as well as spontaneous chemotaxis, increased expression of CCR5, and aberrant receptor dynamics, after RANTES stimulation as observed in SF T cells. CONCLUSION: Synovial fluid T cells highly positive for CCR5 show aberrant characteristics; resistant to RANTES in terms of migration, but responsive in terms of dynamics of CCR5.

Arthritis, Rheumatoid↗
PubMed1986

Histamine levels in human synovial fluid.

Histamine levels have been measured in plasma and synovial fluid (SF) obtained from patients with various arthritides, using a radioenzymatic assay procedure. The concentration of the amine in the plasma of these patients was, in general, found to be considerably greater than that seen in a group of control patients without inflammatory joint disease. SF always contained higher levels of histamine than corresponding plasma samples. The results of our preliminary study suggest that the production of histamine within the synovium may be an important factor in the genesis of joint effusions.

Arthritis↗
PubMed1975

Predominance of synovial fluid lymphocytes in early rheumatoid arthritis.

Evaluation of 158 synovial fluids, mainly from patients with rheumatoid arthritis (RA) and the crystal deposition fiseases (CDD), revealed differences in the differential white blood count and numbers of cytoplasmic inclusion bodies (CIB) present. Lymphocytes and CIB were increased in RA, suggesting a different pathologic mechanism from that of CDD. Lymphocyte-like cells contained CIB implying phagocytic capability. Lymphocytes were the initial predominant cell type in RA of less than six weeks disease duration. Later polymorphonuclear leukocytes increased, possibily representing a secondary non-specific inflammatory response similar to that seen in CDD.

Arthritis↗
PubMed1990

Interleukin-2 inhibitor in rheumatoid arthritis synovial fluid does not inhibit mononuclear cell responses to mitogens.

Synovial fluid (SF) from rheumatoid arthritis (RA) patients were tested for their ability to inhibit the proliferative responses of normal peripheral blood mononuclear cells (PBM) to mitogens and interleukin-2 (IL-2). SF significantly inhibited the responses to concanavalin A (CON A) and phytohaemagglutinin (PHA), but significantly enhanced the responses to IL-2. Similarly, SF mononuclear cells (SFM) were hyporesponsive to CON A and PHA compared with autologous PBM, but hyper-responsive to IL-2. It is concluded that an IL-2 inhibitor in RA SF is unlikely to be the cause of SFM hyporesponsiveness to mitogens.

Arthritis, Rheumatoid↗
PubMed1993

Metalloproteinases, tissue inhibitor, and proteoglycan fragments in knee synovial fluid in human osteoarthritis.

OBJECTIVE: To determine the concentrations of human stromelysin-1, collagenase, tissue inhibitor of metalloproteinases (TIMP), and proteoglycan fragments in knee synovial fluid in patients with injury to the meniscus or anterior cruciate ligament, posttraumatic osteoarthritis, primary osteoarthritis, or pyrophosphate arthritis. METHODS: Synovial fluid samples were collected from patients with knee disease diagnosed arthroscopically and radiologically. Concentrations of stromelysin-1, collagenase, and TIMP-1 were determined by sandwich immunoassay, using monoclonal and polyclonal antibodies. Fragments of cartilage proteoglycan containing the chondroitin sulfate-binding region were determined by immunoassay with a polyclonal antibody. RESULTS: Average concentrations of metalloproteinases, TIMP, and proteoglycan fragments in joint fluid were significantly elevated in patients from all disease groups as compared with volunteers with healthy knees (reference group). Stromelysin concentrations in disease groups averaged 15-45 times that of the reference group. The molar ratios between stromelysin and collagenase varied between 10 and 150. The molar ratio between total stromelysin and free TIMP was 0.5 in the reference group and between 1.6 and 5.3 in the disease groups. CONCLUSION: Stromelysin concentration in joint fluid is a parameter that distinguishes diseased joints from healthy joints, with a sensitivity of 84% and a specificity of 90%. The high concentrations of metalloproteinase relative to TIMP in joint fluid from patients with the conditions studied may be associated with cartilage matrix degradation in these arthritides.

Adult↗
PubMed1984

Elastase activity in serum and synovial fluid of patients with connective tissue disorders.

Sera and synovial fluids (SF) from patients with connective tissue disorders had significantly lower levels of elastase than sera from healthy controls. Patients with rheumatoid arthritis had significantly lower elastase activity in SF compared with serum. Elastase activity in serial serum samples from patients with systemic lupus erythematosus undergoing plasmapheresis showed some relationship with the clinical condition but little correlation with levels of circulating immune complexes. It was concluded that free elastase in the serum or SF was unlikely to be of much pathological significance in connective tissue disease.

Adult↗
PubMed1984

The effect of juvenile inflammatory synovial fluid on in vitro cartilage.

Inflammatory and noninflammatory juvenile synovial fluid (SF) samples were examined in vitro for their effect on cartilage. SF from 3 of 6 patients with pauciarticular juvenile arthritis (JA), 7 of 7 patients with polyarticular JA and 2 of 2 patients with ankylosing spondylitis had significantly increased proteoglycan (PG) releasing activity in 4 day cultures of living but not of freeze-killed cartilage. Noninflammatory SF did not cause increased PG release. Cartilage collagen content was not significantly altered by either inflammatory or noninflammatory SF. Our in vitro results showed PG releasing activity was present in inflammatory but not in normal juvenile SF.

Animals↗
PubMed2001

Expression of matrix metalloproteinase-2 and -9 in synovial fluid of the temporomandibular joint accompanied by anterior disc displacement.

The purpose of this study was to determine whether or not matrix metalloproteinases (MMPs) in synovial fluid are helpful in the biochemical diagnosis of temporomandibular joint (TMJ) disorder (TMD). We examined the synovial fluid from 38 TMD patients with disc displacement and 20 volunteers by gelatin zymography and immunoblotting analysis to clarify the involvement of the joint pathology from the viewpoint of expression of MMPs. Two gelatinolytic enzymes, MMP-2 and -9, were detected in the samples. The incidences of expression, except for pro-MMP-2, in anterior disc displacement (ADD) without reduction (ADD w/o R) were significantly higher than in ADD with reduction (ADD w R) (P<0.05). Quantitative analysis showed that the degree of MMP-2 and -9 expression in ADD w/ o R were higher than in ADD w R. These data suggest that the presence or absence of disc reduction is a major turning point in the process of joint destruction, and these MMPs are useful as biochemical markers for TMD diagnosis.

Adolescent↗
PubMed1986

Methylprednisolone infusion therapy in rheumatoid arthritis patients. The effect on synovial fluid lymphocyte subsets and inflammatory indices.

Paired samples of synovial fluid (SF) and blood were obtained prior to and at 4 and 24 hours following high-dose methylprednisolone infusion therapy in a group of patients with refractory rheumatoid arthritis. After therapy there was a significant decrease in numbers of polymorphonuclear leukocytes, lymphocytes, immune complexes, and C-reactive protein in the SF. Measurement of lymphocyte subsets, using monoclonal antibodies, revealed that at 4 hours postinfusion, there was a disproportionate decrease in the percentage of SF lymphocytes expressing class II antigens (HLA-DR or Ia-like). These data suggest that glucocorticoids induce rapid changes in SF indices of disease activity and may directly influence T cell activation within the rheumatoid joint.

Aged↗
PubMed1992

Treatment with colchicine decreases white cell counts in synovial fluid of asymptomatic knees that contain monosodium urate crystals.

Synovial fluid (SF) samples, all containing monosodium urate crystals, obtained from 18 asymptomatic knees of 12 patients with gout, were serially analyzed before and one month after treatment with 1 mg/day of oral colchicine. Before treatment, the SF contained 612 [confidence limits (CL) = 282, 942] cells/mm3, while after treatment these values decreased to 274 (CL = 73, 471) cells/mm3 (p less than 0.005). In all cases, pretreatment cell counts were higher than the corresponding posttreatment values. The percentage of polymorphonuclear leukocytes also diminished from a pretreatment value of 13.0 (CL = 7, 19) to 4.5 (CL = 2, 7) after colchicine administration (p less than 0.005). These results strongly suggest the presence of baseline subclinical joint inflammation which is reduced by colchicine. The preventive effect of this drug on the occurrence of new attacks may relate to its effect in reducing the baseline inflammation of the asymptomatic joints.

Colchicine↗
PubMed2000

Differences in the concentration of various synovial fluid constituents between the distal interphalangeal joint, the metacarpophalangeal joint and the navicular bursa in normal horses.

As a prerequisite for the identification of navicular disease markers, the concentrations of cartilage oligomeric matrix protein (COMP), total glycosaminoglycans (GAG), hyaluronan, metalloproteinases (MMP) 2 and 9 and total protein were measured in synovial fluid samples obtained from the distal interphalangeal joint (DIP), the metacarpophalangeal joint (MCP) and the navicular bursa of 24 horses. Mean GAG, COMP and total protein levels were significantly higher in the DIP joint and in the navicular bursa compared to the MCP joint. Hyaluronan content was lower. MMP -2 activity was present in all fluids measured and had similar levels in different joints. MMP -9 was present in 42 per cent of MCP joint samples and 58 per cent of DIP joint samples and of navicular bursal samples. In relation to the constituents measured, the composition of navicular bursal fluid was similar to the articular synovial fluids, in particular that obtained from the DIP joint. Correlation between the constituents of DIP joint fluid and navicular bursal fluid obtained from the same legs was statistically significant for all the parameters measured.

Animals↗
PubMed1988

[Study of the synovial fluid in rheumatoid arthritis].

A comparative study of the synovial fluid (SF) composition was conducted in 72 patients with rheumatoid arthritis (RA) with relation to the presence of the rheumatoid factor (RF) and clinical manifestations of this disease. A statistically significant difference in the SF composition (the number of leukocytes, neutrophils and ragocytes) was shown in seropositive and seronegative RA. The content of total protein and glucose in SF did not practically differ in both cases. In the common group of the RA patients correlation was established between the number of neutrophils, the level of total protein, percentage of ragocytes, the level of complement in SF and a degree of general inflammatory activity of disease. A subgroup of RA patients (32%) was identified with the prevalence of lymphocytes, a slight rise of the level of total protein, a low percentage of ragocytes and rather a high level of complement. In 87% of the above patients noticeable osteochondrous destruction (III-IV roentgenological stage) was noted against a background of low inflammatory activity. The data obtained confirmed an important role of SF investigation for the elucidation of the mechanisms of pathogenesis, differential diagnosis and prognosis.

Adult↗
PubMed1987

An analysis of the levels of complement components in the synovial fluid in rheumatic diseases.

A linear relationship between the synovial fluid to serum concentration ratios and log molecular weight was found for six plasma proteins, which are largely synthesized by the liver. Production or utilization of a given protein in the joint can, therefore, be determined by its deviation from the calculated diffusion line. Based on this diffusion model the role of the complement system was investigated in the joint effusions of 48 patients with rheumatoid arthritis (RA), 6 patients with osteoarthritis (OA) and 7 patients with meniscus lesions (ML). Among these three groups quantitative differences were found in the metabolism or utilization of several complement components, based on the fact that the ratios were lower than expected for diffusion of proteins of similar molecular weight. The ratios for the RA group were the lowest. In the three patient groups, results showed increased consumption mainly of C3 and C4 locally in the joint. The existence of a real complement activation in the joints of the three different patient groups was further proved by the elevated levels of C3 breakdown products (C3d). Overall this kind of calculation provides us with a method for studying the role of other proteins which may be important in the inflammatory process of the joint.

Arthritis, Rheumatoid↗
PubMed1989

Characterization of the IL-2-receptor on rheumatoid arthritis synovial fluid T cells.

We studied the hypoproliferative response of synovial fluid (SF) T cells in rheumatoid arthritis (RA) using a mitogenic monoclonal antibody (MoAb) specific for the T-cell antigen receptor-associated CD3 complex. RASF T cells are defective in their proliferative response and in the induction of the Tac (p55) component of the IL-2-receptor (IL-2-R) when stimulated with anti-CD3 monoclonal antibody (MoAb). However, fresh RASF T cells bear demonstrable IL-2-R in cross-linking experiments which are not seen in unstimulated peripheral blood (PB). These receptors are functional since RASF T cells proliferate in response to recombinant IL-2 (rIL-2) better than fresh PB T cells from either normal or RA patients. Scatchard analysis indicates increased (4-fold) numbers of high affinity IL-2-R on (phytohaemagglutinin) PHA-activated RASF T cells as compared with comparably activated RAPB T cells. Phorbol myristate acetate (PMA) induces Tac antigen expression in RASF but does not lead to proliferation. The hyporesponsiveness of RASF T cells does not appear to result from lack of IL-2-R, lack of IL-2-R inducibility, or proliferative potential.

Adult↗
PubMed1997

Cartilage markers in synovial fluid in symptomatic knee osteoarthritis.

OBJECTIVE: To investigate if the relative content of aggrecan and cartilage oligomeric matrix protein (COMP) in synovial fluid lavage samples differs between individuals with knee pain with or without evidence of radiological knee osteoarthritis (joint space narrowing). METHODS: In a community based cohort of 204 individuals aged 35-54 years with chronic (> 3 months duration) knee pain, a subgroup of 45 subjects with radiographic osteoarthritis, grade I according to the Ahlbäck classification, was randomly chosen; 45 individuals, age and sex matched from the same cohort with chronic knee pain but with normal radiographs, served as controls, Knee joint fluid was obtained by a standardised lavage procedure. The concentrations of aggrecan core protein epitopes and a cartilage matrix protein (COMP) were determined by immunoassays and the aggrecan/COMP concentration ratio was calculated. RESULTS: The aggrecan/COMP ratio was higher (P < 0.001) in the group with radiographic osteoarthritis than in the control group. CONCLUSIONS: The higher aggrecan/COMP ratios in osteoarthritis could reflect increased cartilage matrix turnover in osteoarthritis with predominant release of aggrecan fragments. Synovial fluid analysis of cartilage markers holds promise as a useful means of monitoring changes in the cartilage turnover in studies of pathogenic mechanisms in osteoarthritis.

Adult↗