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Diagnostic value of synovial fluid microscopy: a reassessment and rationalisation.

This study is in two parts. In the first synovial fluid from 1892 patients with 14 different arthropathies was examined microscopically. Crystals of different types were identified and the disease distribution of these and various cell types, including several not previously reported in synovial fluid, have been described. These features have been used to derive a series of microscopic diagnostic criteria for each arthropathy. The criteria have been used in the second part of the study to examine synovial fluids from 200 patients without knowledge of any clinical diagnosis. Cytological and clinical diagnoses were compared at the end of the study. Matching diagnoses were made in 71 (35.5%) and a short list of differential diagnoses (based on cytological criteria), which included the clinical diagnosis, was made in a further 43 (21.5%). Of the rest, 63 (31.5%) were correctly described as inflammatory or non-inflammatory and in five (2.5%) no diagnosis could be made. Only in seven cases (3.5%) was an inaccurate (false positive) cytological diagnosis made. The results indicate that synovial fluid microscopy is a potentially more important diagnostic screening test in rheumatological and orthopaedic practice than it would at first appear from published reports.

Adult↗

Serum and synovial fluid levels of chondroitin sulfate in patients with osteoarthritis of the knee joint.

We measured serum and synovial fluid levels of chondroitin sulfate (CS) in patients with osteoarthritis (OA) of the knee joint to clarify whether CS levels differ in various stages of OA, and whether measurement of CS levels is useful in evaluating the pathology of OA. The study population was 117 OA patients and 23 healthy young volunteers. Synovial fluid was obtained from 69 of 117 OA patients. The mean serum level of C4S in all of the OA patients was significantly higher than that in the controls (p< 0.05). However, there was little difference between respective OA stages. A small amount of serum C6S (0.2 to 0.6 nmol/ml) was detected in 31.6% of the OA patients and in 26.1%of the control subjects. The mean serum C0S level in all OA patients was significantly higher than that in the controls (p< 0.01). However no significant difference was found between respective OA stages. As for synovial fluid CS, the C4S level in the early stage of OA tended to be higher than that in the advanced stage. C6S levels showed a markedly decreasing trend with advancing OA stage. C0S was not detected in synovial fluid. The present study demonstrated that serum levels of C4S and C0S in OA patients are elevated compared with normal subjects, and that synovial fluid levels of C4S and C6S may provide useful in assessing the pathology of OA.

Adult↗

Concentration and degradation of hyaluronic acid in knee synovial fluid from carrageenin-induced rabbit arthritis.

The concentration and degradation of hyaluronic acid in the synovial fluid of carrageenin-induced arthritic joints of rabbits was studied. A 0.5-ml volume of 1% lambda-carrageenin was intra-articularly injected three times into a right knee joint, and saline into a left. After 5 d from the last injection, inflammatory changes were observed in the synovial membrane and synovial fluid, but not in the articular cartilage. In the inflammatory synovial fluid, lipid peroxide content, phosphatase activity and cell counts were significantly increased, but the copper concentration was not changed. Concentration of polymeric hyaluronic acid and total hyaluronic acid were determined by high-performance liquid chromatography using gel-permeation columns. Total hyaluronic acid was appreciably decreased in the inflammatory fluid. The polymeric hyaluronic acid determined was 38% of the total hyaluronic acid in the inflammatory fluid and 74% in the control fluid. This suggests that in the inflammatory fluid, molecular weights of hyaluronic acid are distributed in the broader range. The concentration of chondroitin sulphates was similar in both the inflammatory fluid and the control fluid, but the content ratio of chondroitin sulphates to hyaluronic acid was higher in the inflammatory fluid. In the inflamed synovial membrane, synthesis of hyaluronic acid as measured by incorporation of [14C]glucosamine into glycoconjugates was increased by about twice that in the control membrane.

Animals↗

Naproxen kinetics in synovial fluid of patients with osteoarthritis.

1. The kinetics of naproxen in synovial fluid were studied in 407 osteoarthritic outpatients with knee effusion requiring aspiration, following a single 1100 mg oral dose of naproxen sodium. 2. The drug concentration-time profiles were described by a biexponential function. Naproxen entered synovial fluid rapidly, reaching a maximum concentration of 36 mg l-1 (Cmax) at 7.5 h. The first order input rate constant (kOs) was 0.41 +/- 0.15 h-1 with a lag time (tlag) of 0.24 +/- 0.36 h. 3. Elimination from the fluid was slow (t1/2 = 31 +/- 12 h) and appreciable drug concentrations were still measurable (27 mg l-1) after 24 h. 4. During once daily dosing of naproxen sodium, naproxen should accumulate in synovial fluid, a steady-state being achieved within a week of treatment. The predicted accumulation ratio based on trough concentration was 2.4.

Adult↗

Cartilage glycosaminoglycan loss in the acute phase after an anterior cruciate ligament injury: delayed gadolinium-enhanced magnetic resonance imaging of cartilage and synovial fluid analysis.

OBJECTIVE: To examine the glycosaminoglycan (GAG) content in cartilage and that in synovial fluid and determine whether they are associated, in patients with an acute anterior cruciate ligament (ACL) injury. METHODS: Twenty-four patients (14 of whom were male) with a mean age of 27 years (range 17-40 years) were assessed with delayed gadolinium-enhanced magnetic resonance imaging (MRI) of cartilage an average of 3 weeks after an ACL rupture and compared with 24 healthy volunteers. Two hours after an intravenous injection of Gd-DTPA(2-) (0.3 mmoles/kg body weight), quantitative measurements of the T1 relaxation time (T1(Gd) [T1 relaxation time in the presence of Gd-DTPA]) were made in lateral and medial femoral weight-bearing cartilage. In the patients, synovial fluid was aspirated immediately before the MRI, and GAG was analyzed using dye precipitation with Alcian blue. RESULTS: Fifteen of the 24 patients had an isolated bone bruise in the lateral femoral condyle, where the cartilage T1(Gd) was shorter than that in the controls (mean +/- SD 385 +/- 83 msec and 445 +/- 41 msec, respectively; P = 0.004), consistent with decreased GAG content. However, the T1(Gd) was also decreased in the medial femoral cartilage, where bone bruises were rare (376 +/- 76 msec in patients versus 428 +/- 38 msec in controls; P = 0.006). The mean +/- SD synovial fluid GAG concentration in patients was 157 +/- 86 mug/ml and showed a positive correlation with the T1(Gd) (r = 0.49, P = 0.02). CONCLUSION: This study indicates that an ACL injury causes posttraumatic edema of the lateral femoral cartilage but initializes a generalized biochemical change within the knee that leads to GAG loss from both lateral and medial femoral cartilage. In cartilage with a high GAG content (long T1(Gd)), more GAG is released into the synovial fluid, suggesting that cartilage quality is a factor to consider when interpreting cartilage biomarkers of metabolism.

Adolescent↗

Induction of arthritic processes by synovial fluids of rheumatoid arthritis patients in long-term experiments with mice.

Diluted synovial fluids derived from three patients with rheumatoid arthritis (RA) and applied subcutaneously once as a single dose in Swiss mice caused arthritic processes after a long period of latency (10 months). The arthritis-inducing effect was enhanced by storing the original synovial fluids at 4 degrees C for three months. The effects were followed histologically for at least 18 months. In contrast to the theory that RA is based on immunological processes, these preliminary observations support the finding of other authors that a transmissible agent which is rather slow-acting exists in the synovial fluid of RA patients.

Animals↗

Vitamin B12 binding proteins (transcobalamin and haptocorrin) in serum and synovial fluid of patients with rheumatoid arthritis and traumatic synovitis.

The vitamin B12 (cobalamin) binding proteins, transcobalamin and haptocorrin, were measured in serum and synovial fluid of 12 patients with rheumatoid arthritis (RA) and 11 patients with traumatic synovitis (TS). Levels of both the cobalamin-binding proteins of synovial fluid were markedly elevated in RA compared with TS. In serum, the haptocorrin concentrations were significantly increased in RA vis-à-vis TS. In synovial fluid of RA patients, the transcobalamin and haptocorrin concentrations were positively correlated to the concentrations of macrophages and polymorphonuclear granulocytes, respectively. The demonstration of increased synovial fluid transcobalamin/total protein and haptocorrin/total protein ratios in RA vis-à-vis TS provides further evidence of a local production or liberation of the cobalamin-binding proteins in synovial fluid of patients with RA.

Arthritis, Rheumatoid↗

Synovial fluid leukocyte apoptosis is inhibited in patients with very early rheumatoid arthritis.

Synovial leukocyte apoptosis is inhibited in established rheumatoid arthritis (RA). In contrast, high levels of leukocyte apoptosis are seen in self-limiting crystal arthritis. The phase in the development of RA at which the inhibition of leukocyte apoptosis is first apparent, and the relationship between leukocyte apoptosis in early RA and other early arthritides, has not been defined. We measured synovial fluid leukocyte apoptosis in very early arthritis and related this to clinical outcome. Synovial fluid was obtained at presentation from 81 patients with synovitis of < or = 3 months duration. The percentages of apoptotic neutrophils and lymphocytes were assessed on cytospin preparations. Patients were assigned to diagnostic groups after 18 months follow-up. The relationship between leukocyte apoptosis and patient outcome was assessed. Patients with early RA had significantly lower levels of neutrophil apoptosis than patients who developed non-RA persistent arthritis and those with a resolving disease course. Similarly, lymphocyte apoptosis was absent in patients with early RA whereas it was seen in patients with other early arthritides. The inhibition of synovial fluid leukocyte apoptosis in the earliest clinically apparent phase of RA distinguishes this from other early arthritides. The mechanisms for this inhibition may relate to the high levels of anti-apoptotic cytokines found in the early rheumatoid joint (e.g. IL-2, IL-4, IL-15 GMCSF, GCSF). It is likely that this process contributes to an accumulation of leukocytes in the early rheumatoid lesion and is involved in the development of the microenvironment required for persistent RA.

Adult↗

Co-expression of interleukin-1beta and tumor necrosis factor alpha in synovial tissues and synovial fluids of temporomandibular joint with internal derangement: comparison with histological grading of synovial inflammation.

BACKGROUND: It has been clarified that interleukin-1 (IL-1)beta and tumor necrosis factor (TNF)alpha play an important role in pathogenesis of various joint disease. The purpose of this study was to investigate the cellular source of IL-1beta and TNFalpha in temporomandibular joint (TMJ), and to analyze the relation between the expression of these cytokines and the intensity of TMJ synovial inflammation. METHODS: We examined 33 synovial biopsy specimens from patients with internal derangement of the TMJ by an immunohistochemical technique using specific antibodies to IL-1beta and TNFalpha. We also studied 20 synovial fluids from the patients by enzyme-linked immunosorbent assay method. These data are compared with histological grading of synovial inflammation by Gynther's system. RESULTS: Both IL-1beta and TNFalpha were predominantly localized in the synovial lining cell layer and the blood vessels of synovial biopsy specimens obtained from patients with TMJ internal derangement. A statistically significant correlation was found between the intensity of IL-1beta expression and that of TNFalpha. Additionally, the intensity of TNFalpha expression was statistically correlated with histological grading by Gynther's system. CONCLUSION: These results supported that IL-1beta and TNFalpha may be involved in the occurrence of TMJ internal derangement and that they coordinately play an role in pathogenesis of TMJ internal derangement.

Adult↗

[IgG RF analysis of synovial fluid of the knee joints in 35 patients with rheumatic disease].

Rheumatoid arthritis (RA) is an autoimmune disease. IgG RF in the synovial fluid is a significant indicator for the pathogenesis and differential diagnosis of this disease. To now, RF tested by latex agglutination belongs to the IgM type whereas, in the rheumatoid synovial fluid, most of immunocomplex molecules are IgG which bind into a complex. IgG RF was determined by ELISA in the synovial fluid of 35 cases of joint disorders, including 12 cases of RA, 7 cases of reactive arthritis (ReA), 5 cases of osteoarthritis (OA) and 11 cases of non-synovitis (non-S) which were regarded as negative control. The upper limit of OD value +2s was 0.5. Using this standard 7 out of 12 cases of RA were positive. 2 of 5 cases of OA were weakly positive, and only one case of ReA was positive. This suggest that this test is valuable in the diagnosis of RA. We have also observed the relation between IgG RF in synovial fluid and X-ray and arthroscopic findings of the same knee joint. In 5 of 7 cases of positive RA, II-III X-ray changes and II-III cartilage destruction arthroscopic findings were noted, suggesting a relationship between IgG RF in synovial fluid and articular damage.

Adolescent↗

Detection of interleukin 8 biological activity in synovial fluids from patients with rheumatoid arthritis and production of interleukin 8 mRNA by isolated synovial cells.

The presence of neutrophils in the synovial joint of patients with rheumatoid arthritis (RA) is thought to be due to the activity of chemotactic factors released by activated cells in the joint. We have shown in this report, for the first time, the abundance of one such factor, interleukin 8 (IL 8), in the synovial fluid of patients both with RA and other non-RA joint diseases, and the spontaneous production of IL 8 mRNA by RA synovial cells in culture. There was no correlation between the levels of chemotactic activity and IL 8 protein, suggesting that other factors with similar neutrophil chemotactic activity are also present in the synovial fluid exudate. In support of this concept neither the level of chemotactic activity nor IL 8 protein levels correlated with neutrophil or leukocyte infiltration, indicating that the mechanism of migration into the inflammatory environment of the joint is complex. Such migration is likely to be due to a number of chemotactic signals in addition to IL 8, which may either synergize with, or inhibit, the action of IL 8.

Adult↗

[Immunological findings in serum and synovial fluid in patients with rheumatoid arthritis (author's transl)].

Sera and synovial fluid were investigated in 45 patients with rheumatoid Arthritis and 50 patients with osteoarthritis in inflammatory exacerbation (control group). The following tests were performed: IgG, IgM, IgA determinations, complement components C3, C3, C4, C3-proactivator, ceruloplasmin, electrophoresis, LDH and total acid phosphatase. 1. Serum levels of the ceruloplasmin, alpha 1, alpha 2 and gamma fractions of electrophoresis are significantly higher in patients with rheumatoid arthritis than in patients with osteoarthritis. 2. Synovial fluid: a) There is a significantly higher concentration of IgG, IgM, IgA, C3-proactivator and total acid phosphatase in the synovial fluid of patients with rheumatoid arthritis. b) C4 is significantly lower in patients with rheumatoid arthritis. c) Both groups were also compared with the help of a point system. Every patient received a plus point when the following criteria were seen: IgM greater than 150 mg/100 ml, C3 greater than 50 mg/100 ml, ceruloplasmin greater than 35 mg/100 ml, alpha 1 greater than 0.21 g%, alpha 2 greater than 0.44 g%, beta greater than 0.60 g% and gamma fraction on electrophoresis greater than 0.90 g%. Another point was added if the criteria ceruloplasmin greater than 22 mg/100 ml and C4 less than 17 mg/100 ml were simultaneously seen. With the help of this points system 48 out of the 50 osteoarthritis patients (96%) received zero points, one received 1 point and one 2 points, as opposed to the patients with rheumatoid arthritis where 35 out of 45 (78%) received one or more points. d) The differentation is not improved through additional testing of the rheumatic factors.

Acid Phosphatase↗

Structure of T cell antigen receptor beta chain in synovial fluid cells from patients with rheumatoid arthritis.

We attempted to identify a clonal proliferation of T cells from synovial fluid samples from patients with rheumatoid arthritis, using techniques of restriction fragment length polymorphism. We used probes for the beta chain of the T cell receptor to analyze restriction fragments prepared from the genomic DNA of synovial fluid mononuclear cells from 10 patients and synovial fluid T cell preparations from 5 additional patients. The results demonstrated unarranged (germline) T cell receptor gene fragments of DNA in all cell preparations, indicating the lack of clonality of rheumatoid arthritis synovial fluid T cells.

Arthritis, Rheumatoid↗

Activation of progelatinase B in synovial fluids of patients with rheumatoid arthritis, with reference to stromelysin-1 and tissue inhibitor of matrix metalloproteinase-1.

OBJECTIVE: To clarify whether stromelysin-1 and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are involved in the modulation of activation of progelatinase B in the synovial fluid of patients with rheumatoid arthritis (RA). METHODS: Gelatinases in the synovial fluid of patients with RA were analyzed by gelatin zymography. Concentrations of stromelysin-1 and TIMP-1 were measured using a specific sandwich enzyme linked immunosorbent assay. RESULTS: Forty-three rheumatoid synovial fluids containing progelatinase B were examined to clarify whether the enzyme was activated by incubation. Incubation at 37 degrees C caused the conversion of progelatinase B to the active form in 22 of the 43 synovial fluids. The levels of both stromelysin-1 and TIMP-1 were determined for each group and the concentration ratio of stromelysin-1/TIMP-1 in the synovial fluids of each group was highly correlated to the activation of progelatinase B. CONCLUSION: The balance between the concentrations of stromelysin-1 and of TIMP-1 in the synovial fluid appears to determine whether the progelatinase B molecule causes conversion into the active form.

Arthritis, Rheumatoid↗

The characterisation and function of the polysaccharidases of human synovial fluid in rheumatoid and osteoarthritis.

A potential enzymic mechanism for the degradation of glycosaminogly cans was characterised using enzymes found in rheumatoid synovial fluid from the knee joint. This mechanism involves a true hyluronidase together with the concerted action of beta-glucuronidase and beta-N-acetylhexosaminidase. The contribution of the exopolysaccharidases to hyaluronate degradation was demonstrated by the use of specific inhibitors, while the distinct identity of a true hyaluronidase was shown by ammonium sulphate and agarose gel column fractionations. Only the hyluronidase fraction was capable of degrading high molecular weight hyaluronate. The exopolysaccharidase activities were shown to be markedly elevated in rheumatoid as compared to osteoarthritic synovial fluid and also normal serum. On the other hand, hyluronidase was similarly active in rheumatoid and osteoarthritic synovial fluids; both these levels were lower than that of normal human serum. Hyaluronidase in synovial fluid may thus be derived by diffusion from serum, since it is of relatively low molecular weight (60 000). The pH requirements of this enzyme system and the strong inhibition of hyaluronidase by synovial fluid make it unlikely that the mechanism operates extracellularly. It is proposed that as a lysosomal mechanism, however, it is an important contributing factor in the chronic erosion process characteristic of rheumatoid arthritis.

Acetates↗

Cartilage-derived biomarkers of osteoarthritis in synovial fluid of dogs with naturally acquired rupture of the cranial cruciate ligament.

OBJECTIVE: To compare synovial fluid biomarkers of cartilage metabolism in joints with naturally acquired or experimentally induced cranial cruciate ligament (CCL) rupture and determine correlations with stage and severity of disease in dogs. ANIMALS: 95 dogs with ruptured CCL, 8 dogs with experimentally ruptured CCL, and 24 healthy dogs. PROCEDURES: Synovial fluid was assayed for chondroitin sulfate neo-epitopes 3B3(-) and 7D4 and glycosaminoglycan (GAG) concentration. Results were correlated with demographic data, duration of lameness, radiographic osteoarthritis score, and intra-articular lesions. RESULTS: The 7D4 concentrations and 7D4:GAG in synovial fluid from joints with naturally acquired CCL rupture and experimental CCL transection were similar and significantly greater than values for healthy control joints. The 3B3(-) concentrations in the CCL-deficient groups were not significantly different, although only values in the naturally acquired CCL rupture group were significantly greater than those in the healthy control group. Within the naturally acquired CCL rupture group there was a significant correlation between 3B3(-) and 7D4 concentrations. However, there were no significant correlations between biomarker concentrations and continuous demographic or disease-related variables or differences in biomarker concentrations with different categories of disease. CONCLUSIONS AND CLINICAL RELEVANCE: Synovial fluid biomarker concentrations were significantly increased in joints with secondary osteoarthritis associated with naturally acquired or experimental CCL rupture; however, lack of apparently simple relationships with demographic variables or stage or severity of disease limits their clinical usefulness.

Animals↗

Measurement of synovial fluid and serum concentrations of the 846 epitope of chondroitin sulfate and of carboxy propeptides of type II procollagen for diagnosis of osteochondral fragmentation in horses.

OBJECTIVE: To determine whether serum or synovial fluid concentrations of chondroitin sulfate epitope 846 and carboxy propeptides of type II collagen (CPII) can be used to diagnose osteochondral fragmentation (OC) in horses. ANIMALS: 38 horses with unilateral OC of the radiocarpal (n = 31) or intercarpal (33) joints and 8 clinically and radiographically normal horses. Procedures-For horses with OC, serum and synovial fluid concentrations of epitope 846, CPII, and keratan sulfate (KS) were determined, along with synovial fluid WBC counts and total protein concentrations. Serum epitope 846, CPII, and KS concentrations were measured in control horses. RESULTS: Synovial fluid epitope 846 and total protein concentrations were significantly higher in the joints with OC than in unaffected joints, but CPII and KS concentrations and WBC counts were not. Synovial fluid total protein and 846 epitope concentrations were linearly related to grade of OC. Serum epitope 846 and CPII concentrations were significantly higher in horses with OC than in control horses. Discriminant analysis allowed 27 of 34 (79%) horses to be correctly classified as having or not having OC on the basis of serum epitope 846 and CPII concentrations. CONCLUSIONS: Results suggest that serum and synovial fluid concentrations of epitope 846 and CPII are associated with OC. Increases in concentrations of epitope 846 and CPII suggest that increased synthesis of cartilage aggrecan and type II procollagen may be associated with OC. CLINICAL RELEVANCE: Measurement of serum epitope 846 and CPII concentrations may be useful in the diagnosis of OC in horses.

Animals↗

Effects of non-steroidal anti-inflammatory drugs and prednisolone on synovial fluid white cells, prostaglandin E2, leukotriene B4 and cyclic AMP in patients with rheumatoid arthritis.

Altogether 53 patients (31 women, 22 men) with definite rheumatoid arthritis were randomly divided into groups of 5-6 patients and treated for one day only with one of the following non-steroidal anti-inflammatory drugs (NSAIDs): acetylsalicylic acid, carprofen, diclofenac, indomethacin, naproxen, proquazone, timegadine, tolfenamic acid or paracetamol, and with prednisolone, in recommended doses. Synovial fluid samples were collected before and after the treatment. White cell count and its differentiation as well as the concentrations of protein, cyclic adenosine-3',5'-monophosphate (cAMP), prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) were measured from the synovial fluid. Synovial fluid leukocyte counts correlated with PGE2 concentrations, but showed no correlation with LTB4 levels before treatment. Significant changes were seen in the form of lowered PGE2 values following treatment with the clinically and experimentally most potent NSAIDs, and as depressed LTB4 levels following prednisolone treatment. The other markers of inflammation are obviously more resistant, changing only slowly during prolonged treatment, and may thus be, at least in part, secondary to the changes in prostanoids.

Anti-Inflammatory Agents, Non-Steroidal↗