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Effects of human knee synovial fluid on chondrogenesis in vitro.

To evaluate the healing environment of chondral defects, synovial fluid samples from knees of 25 patients with a traumatic chondral defect were evaluated. Effect of the synovial fluid on chondrogenesis was measured using a chick limb bud bioassay. Samples from 11 of 17 acutely injured knees stimulated chondrogenesis, samples from 4 were inhibitory, and samples from 2 showed no effect. Samples from six of eight chronically injured knees inhibited chondrogenesis; the other two samples stimulated chondrogenesis. The relationship between acute versus chronic injury and effect of synovial fluid on chondrogenesis was statistically significant. We conclude that synovial fluid contains factors that stimulate cartilage healing in the acute period following traumatic injury, but this effect can become inhibitory if the lesion is allowed to become chronic.

Adult↗

[Alizarin red S staining of calcium compound crystals in synovial fluid].

In order to study the clinical value of alizarin red S staining of calcium compound crystals in synovial fluid, the optimum conditions for staining and the detectable threshold of the crystals were examined. The staining of crystals was mainly affected by the concentration of the dye and pH of the solution. The best results were obtained with an alizarin red S concentration of 1.5-3.0% for hydroxyapatite (HA) and 2.0-3.0% for calcium pyrophosphate dihydrate (CPPD) crystal. The optimal pH of the solution was 4.0-6.0 for HA and 4.0-5.5 for CPPD crystal. For the detection of crystals phagocytosed by synovial fluid leukocytes, staining was enhanced by the addition of chloral hydrate to the dye solution which increased the permeability of the cell membrane. The detectable threshold levels of HA and CPPD crystal in synovial fluid by alizarin red S staining were 0.1 micrograms/ml and 0.5 micrograms/ml, respectively. These results suggest that alizarin red S staining is suitable for screening calcium compound crystals in synovial fluid, because it is more sensitive than other methods, such as polarized microscopy and X-ray diffraction. Alizarin red S staining was performed on 148 synovial fluids from patients with various joint diseases. The staining was positive in 100% of synovial fluids from patients with CPPD deposition disease, in 54% of fluids with osteoarthritis, and in 39% of fluids with rheumatoid arthritis. In osteoarthritis, the increase in the proportion of positive cases was found to be in accordance with the radiological grading of the joints.

Anthraquinones↗

Total and free ketoprofen in serum and synovial fluid after intramuscular injection.

Free and total ketoprofen levels in serum and synovial fluid were determined in 37 patients after a single intramuscular injection of ketoprofen, 100 mg. Free drug was separated by equilibrium dialysis. Ketoprofen was assayed by HPLC. Ketoprofen penetrated into the joints rapidly and significant concentrations were found at 15 minutes. The equilibrium time was about 3 1/2 hours. The AUC for total ketoprofen was greater in serum than in synovial fluid. On the other hand, the free fraction AUC in the serum and synovial fluid were quite similar. The mean residence time in the joint was about three times that in the systemic circulation. Ketoprofen was strongly bound to proteins and the percentage of free ketoprofen was not significantly different between serum and synovial fluid. These results provide a possible explanation for duration of the therapeutic effect of ketoprofen despite the short elimination half-life from the serum.

Adolescent↗

Apparent viscosity of the synovial fluid from mid-carpal, tibiotarsal, and distal interphalangeal joints of horses.

OBJECTIVE: To compare the apparent viscosity of normal synovial fluid of the mid-carpal, tibiotarsal, and interphalangeal joints of horses. DESIGN: Viscosity evaluation over a range of shear rates was used to characterize the apparent viscosity of synovial fluids from the 3 joints. ANIMALS: 60 clinically normal adult horses. PROCEDURE: Viscosity data for synovial fluid samples were obtained over a shear rate range of 10 to 250/s and apparent viscosity was calculated at 50, 100, 150, 200, and 250/s. Effect of shear rate on apparent viscosity was determined, using a two-way ANOVA, with significant differences tested, using a Tukey's test at a significance level of P < 0.05. RESULTS: Synovial fluid from all these joints indicated shear thinning behavior: decreased apparent viscosity with increased shear rate. Apparent viscosity of synovial fluid from the 3 joints was not significantly different over the shear rate range of 50 to 250/s. CONCLUSION: Results of this study indicate that the apparent viscosity of the distal interphalangeal joint is not less than that of other joints, as has been reported. CLINICAL RELEVANCE: The observation of decreased synovial fluid viscosity of distal interphalangeal joint fluid should be considered as suggestive of joint disease.

Analysis of Variance↗

Anti-Sm and anti-DNA antibodies in paired serum and synovial fluid samples from patients with SLE.

Antibodies to nuclear antigens (ANAs) are frequently found in the serum of patients with connective tissue diseases (CTDs). Particularly systemic lupus erythematosus (SLE), and have been implicated in the immune-complex mediated pathogenesis of these diseases. In this study we have compared the occurrence of precipitating ANAs in paired samples of serum and synovial fluid from patients with different CTDs. Of the 30 patients examined 3 had precipitating ANAs in their serum only, 1 in the synovial fluid only, and 3 had antibodies in both serum and synovial fluid. Precipitating ANAs in synovial fluid were found in 3/6 patients with SLE, 1 patient with RA/Sjogren's syndrome overlap, and one patient with RA/SLE overlap. Of the other 15 patients with RA, 2 had precipitating antibodies only in their serum. Two of the SLE patients had anti-Sm antibody, one in serum only and the other in both serum and synovial fluid. Detection by ELISA of class specific anti-Sm antibodies in serum or synovial fluid paralleled the occurrence of antidenatured DNA antibodies when both specificities occurred together. One SLE patient did show evidence in synovial fluids of elevated concentrations of specific antibody classes to individual antigens; however, elevated levels were more frequently found in serum. Local production of ANAs does not, therefore, appear to be a feature of synovial fluids from SLE patients.

Antibodies↗

Lysozyme concentrations in synovial fluid, pleural fluid and thoracic duct lymph in rheumatoid arthritis.

Lysozyme (LZM) concentrations in synovial fluid were determined in patients with seropositive and seronegative rheumatoid arthritis (RA) and in patients whose arthritic exudates had been caused by Reiter's disease, a Yersinia enterocolitica infection, osteoarthritis, or trauma. Patients with rheumatoid disease had significantly higher levels of lysozyme in synovial fluid than patients with non-rheumatic diseases. The concentration of lysozyme correlated with the number of polymorphonuclear leukocytes in synovial fluid in seronegative--but not in seropositive--rheumatoid arthritis. In patients with rheumatic arthritis the lysozyme level correlated inversely with the concentration of glucose in synovial fluid. In patients with rheumatoid pleural effusion, lysozyme levels in pleural fluid were comparable to those in serum. The concentration of LZM in thoracic duct lymph was roughly the same as in serum. During drainage of thoracic duct lymph, the lysozyme level in serum decreased.

Adult↗

[An immunochemical study of extracts from synovial fluid cells in rheumatoid arthritis].

Extracts were obtained by ultrasonic vibration from synovial fluid cells of patients with rheumatoid arthritis and rabbit antisera against the extracts were prepared. The antisera were absorbed with normal human plasma. Double immunodiffusion revealed two precipitin lines between the extracts and the antisera. These antigens were called P1 and P2. P1 and P2 antigens were detected in 17 (94%) and 10 (56%), respectively, out of 18 rheumatoid synovial fluids. In osteoarthritic fluids, P1 antigen was detected in only one (4%) of 25 cases but P2 antigen was never detected. The P1 antigen was not inactivated by trypsin or deoxyribonuclease but P2 antigen was digested by trypsin suggesting its protein character. Though it is not clear that these antigens are specific for rheumatoid arthritis, these antigens in synovial fluid cells seem to be associated with the rheumatoid synovial fluid.

Adult↗

Peripheral blood but not synovial fluid natural killer T cells are biased towards a Th1-like phenotype in rheumatoid arthritis.

Natural killer T (NKT) cells have been implicated in the regulatory immune mechanisms that control autoimmunity. However, their precise role in the pathogenesis of rheumatoid arthritis (RA) remains unclear. The frequency, cytokine profile and heterogeneity of NKT cells were studied in peripheral blood mononuclear cells (PBMCs) from 23 RA patients and 22 healthy control individuals, including paired PBMC-synovial fluid samples from seven and paired PBMC-synovial tissue samples from four RA patients. Flow cytometry revealed a decreased frequency of NKT cells in PBMCs from RA patients. NKT cells were present in paired synovial fluid and synovial tissue samples. Based on the reactivity of PBMC-derived NKT cells toward alpha-galactosylceramide, RA patients could be divided into responders (53.8%) and nonresponders (46.2%). However, NKT cells isolated from synovial fluid from both responders and nonresponders expanded upon stimulation with alpha-galactosylceramide. Analysis of the cytokine profile of CD4+ and CD4- PBMC derived NKT cell lines from RA patients revealed a significantly reduced number of IL-4 producing cells. In contrast, synovial fluid derived NKT cell lines exhibited a Th0-like phenotype, which was comparable to that in healthy control individuals. This suggests that synovial fluid NKT cells are functional, even in patients with nonresponding NKT cells in their blood. We conclude that, because the number of Valpha24+Vbeta11+CD3+ NKT cells is decreased and the cytokine profile of blood-derived NKT cells is biased toward a Th1-like phenotype in RA patients, NKT cells might be functionally related to resistance or progression of RA. Providing a local boost to the regulatory potential of NKT cells might represent a useful candidate therapy for RA.

Adult↗

Assessment of the effects of age and joint disease on hydroxyproline and glycosaminoglycan concentrations in synovial fluid from the metacarpophalangeal joint of horses.

OBJECTIVE: To assess the effects of age and joint disease on hydroxyproline and glycosaminoglycan (GAG) concentrations in synovial fluid from the metacarpophalangeal joint of horses and evaluate the association of those concentrations with severity of osteoarthritis and general matrix metalloproteinase (MMP) activity. SAMPLE POPULATION: Synovial fluid was collected from the metacarpophalangeal joints of foals at birth (n = 10), 5-month-old foals (10), 11-month-old foals (5), and adult horses (73). PROCEDURE: Hydroxyproline and GAG concentrations were determined in synovial fluid samples. The severity of osteoarthritis in adult joints was quantified by use of a cartilage degeneration index (CDI) and assessment of general MMP-activity via a fluorogenic assay. RESULTS: Hydroxyproline and GAG concentrations in synovial fluid were highest in neonates and decreased with age. Concentrations reached a plateau in adults by 4 years and remained constant in healthy joints. In synovial fluid from osteoarthritic joints, hydroxyproline and GAG concentrations were not increased, compared with unaffected joints, but hydroxyproline were significantly correlated with the CDI and general MMP activity. There was no significant correlation between GAG concentration and CDI value or MMP activity. CONCLUSIONS AND CLINICAL RELEVANCE: Changes in hydroxyproline concentration in synovial fluid appeared to indicate damage to collagen of the articular cartilage. In joints with osteoarthritis, the lack of high GAG concentration in synovial fluid and the absence of a significant correlation between GAG concentration and CDI values or MMP activity may severely limit the usefulness of this marker for monitoring equine joint disease.

Age Factors↗

Alteration of tryptophan metabolism in the synovial fluid of patients with rheumatoid arthritis and osteoarthritis.

Tryptophan, its metabolites and related enzyme activity in synovial fluid, blood and urine in patients with rheumatoid arthritis (RA) and osteoarthritis (OA) were measured. The levels of tryptophan, kynurenine and anthranilic acid in the synovial fluid higher in RA were than in OA, whereas the xanthurenic acid level was equal in RA and OA. Indoleamine 2, 3-dioxygenase activity in the synovial membrane was higher in RA than in OA. 5-Hydroxytryptamine (5-HT) levels in the synovial fluid and the blood and the 5-hydroxyindole acetic acid (5-HIAA) level in the synovial fluid were essentially the same for both diseases. However, the 5-HIAA level in the serum of RA patients was higher than in those with OA, and the 5-HIAA level in the urine of RA patients was lower than in those with OA. In addition, monoamine oxidase-A and B activity in the synovial fluid of RA patients was decreased than in those with OA. These findings suggest that metabolism of tryptophan is altered in patients with RA.

Arthritis, Rheumatoid↗

Lymphocytes in rheumatoid and nonrheumatoid synovial fluids. Nonspecificity of high T-cell and low B-cell percentages.

Lymphocytes were studied in paired peripheral blood and synovial fluid samples from patients with various forms of arthritis, including rheumatoid arthritis (group I) and other polyarthritides of unknown origin (group II), as well as arthritides generally considered not to be immunologically mediated, such as crystal synovitis, traumatic arthritis, osteoarthrosis, and pigmented villonodular synovitis (group III). In all 3 groups the percentages of T lymphocytes were significantly higher in synovial fluids than in the peripheral blood, whereas those of the synovial fluid B lymphocytes were consistently very low and occasionally nil. Absolute numbers of synovial fluid lymphocytes were significantly higher in groups I and II as compared with group III, and in the peripheral blood absolute numbers of lymphocytes in groups I and II were significantly lower than in controls. No correlation was found between absolute numbers of lymphocytes and complement activity in the synovial fluid. The characteristic pattern of high T-cell and very low B-cell percentages in synovial fluids is a general feature of inflammatory exudates and cannot be considered an expression of cell-mediated immunity in itself.

Arthritis↗

Plasma pharmacokinetics and synovial fluid concentrations after oral administration of single and multiple doses of celecoxib in greyhounds.

OBJECTIVE: To determine the plasma pharmacokinetics and synovial fluid concentrations after oral administration of single and multiple doses of celecoxib in Greyhounds. ANIMALS: 7 adult Greyhounds. PROCEDURES: Dogs received celecoxib (median dose, 11.8 mg/kg [range, 11.5 to 13.6 mg/kg], PO, q 24 h) for 10 days. Blood samples were collected prior to administration of celecoxib and serially for 24 hours after the 1st and 10th doses were administered. A synovial joint catheter was placed into a stifle joint in each dog for collection of synovial fluid samples. Concentrations of celecoxib in plasma and synovial fluid were quantified by use of a validated liquid chromatography/mass spectrometry method. Identification of hydroxy- and carboxyl-celecoxib in plasma and synovial fluid was also performed. Pharmacokinetic parameters were determined by use of noncompartmental analysis. RESULTS: Administration of multiple doses of celecoxib resulted in a significant decrease (40%) in median area under the curve (AUC) values and a corresponding decrease in median maximum concentrations (Cmax; 2,620 to 2,032 ng/mL) between the 1st and 10th doses. Synovial fluid concentrations were less than the corresponding plasma concentrations at all times except 24 hours after administration of the 10th dose of celecoxib. CONCLUSIONS AND CLINICAL RELEVANCE: Celecoxib distributes into the synovial fluid of Greyhounds. Although the exact mechanism for the decreases in AUC and Cmax is not known, results suggested that the plasma pharmacokinetics of celecoxib are different after administration of multiple doses in Greyhounds. These findings warrant further investigation on the absorption, distribution, metabolism, and elimination of celecoxib in Greyhounds and other breeds of dogs.

Animals↗

Platelets in the synovial fluid of patients with rheumatoid arthritis.

In a study of synovial fluid from 110 patients with various forms of arthritis, platelets were identified in the synovial fluid of all the 50 rheumatoids, in 18 out of the 25 (72%) with osteoarthritis and in all 35 of those with other forms of inflammatory osteoarthrosis. Identification of platelets by light microscopy was confirmed by electron microscopy. Platelet counts were significantly higher in rheumatoid fluid (mean 14 988/mm3; range 1000-65 000/mm3) compared with fluid from patients with osteoarthrosis (mean 1 592/Mm3; 0-10 000/mm3). In addition, significantly higher platelet counts were found in the synovial fluid (SF) of inflamed joints. There was a positive correlation between the SF platelet count and the total white cell count, polymorph count, hydrogen ion concentration, knee score, acid phosphatase and 5-nucleotidase activity and a negative correlation with the glucose level. All these factors indicate joint activity. Finally, platelet numbers correlated with SF levels of immunoglobulin M, and seropositive patients had significantly higher platelet counts in the SF compared with seronegative patients. Rheumatoid patients with thrombocytosis also had higher SF platelet counts. The close relationship of the SF platelet count to other indices of inflammation supports the concept that platelets may directly contribute to synovial inflammation by a variety of pathways.

Arthritis↗

Comparison of the concentrations of pentosidine in the synovial fluid, serum and urine of patients with rheumatoid arthritis and osteoarthritis.

OBJECTIVE: Pentosidine, an advanced glycation end product (AGE), has recently been observed to be elevated in rheumatoid arthritis (RA). The aim was to elucidate which pentosidine levels, i.e. in serum, synovial fluid or urine, are more related to the disease status of RA. METHODS: We measured levels of pentosidine in serum, synovial fluid or urine in RA compared with osteoarthritis (OA), and examined the relationship between pentosidine and RA disease activity. Subjects were 20 patients with RA and 22 patients with OA. RESULTS: In total RA and OA patients combined, there was a significant correlation between pentosidine in serum, synovial fluid and urine. Pentosidine in serum and synovial fluid was significantly higher in RA than in OA. In RA, there were significant correlations between pentosidine in serum and synovial fluid and C-reactive protein, Lansbury index (LI) and erythrocyte sedimentation rate. CONCLUSIONS: These results demonstrate that pentosidine levels in body fluids correlated with each other, and pentosidine in serum and in synovial fluid is associated with the systemic inflammatory activity of RA. Higher or similar concentrations of pentosidine in serum compared with synovial fluids indicate that the elevated pentosidine levels in serum in RA are not derived from the synovial fluid, but from an increase in the formation of pentosidine in the whole body in RA. Among body fluids, serum pentosidine was the superior indicator for RA disease status.

Adult↗

In vitro mitogen stimulation of synovial fluid lymphocytes from rheumatoid arthritis and juvenile rheumatoid arthritis patients: dissociation between the response to antigens and polyclonal mitogens.

The in vitro responses to mitogens of synovial fluid lymphocytes obtained from eight patients with rheumatoid arthritis (RA) and eight patients with juvenile rheumatoid arthritis (JRA) were studied. The results were compared to the transformation of the patient's peripheral blood lymphocytes stimulated with the same mitogens. Both RA and JRA synovial fluid lymphocytes showed a low transformation to the polyclonal mitogens PHA and PWM with a low ratio PHA-response/PWM-response. The stimulatory effect of purified protein derivative of tuberculin (PPD) was high, whereas a Candida albicans antigen preparation gave a more variable stimulation of the synovial fluid lymphocytes. In some patients the complete mitogen transformation profile of lymphocytes obtained from synovial fluid, synovial tissue and peripheral blood was studied. The results of the synovial fluid and tissue lymphocytes were similar.

Antigens↗

Differential expression in rheumatoid synovium and synovial fluid of alpha 4 beta 7 integrin. A novel receptor for fibronectin and vascular cell adhesion molecule-1.

T lymphocyte adhesion to vascular endothelium plays an important role in the immunopathogenesis of rheumatoid arthritis. The migration of T lymphocytes into the synovium is mediated by a variety of adhesion molecules, notably the integrins. We have prepared Act I, a murine mAb that identifies a novel integrin termed alpha 4 beta 7. The natural ligands for alpha 4 beta 7 are vascular cell adhesion molecule-1 and fibronectin; both molecules are up-regulated in the rheumatoid synovium. We investigated the expression of alpha 4 beta 7 in the three compartments of rheumatoid arthritis, the peripheral blood, synovial fluid, and synovial membrane, utilizing the FACS and immunoperoxidase microscopy of frozen tissues. The results of our experiments show a striking differential expression of alpha 4 beta 7 integrin in rheumatoid arthritis. Sixty-two percent of synovial membrane T cells expressed high density alpha 4 beta 7, in contrast to only 4.7% of synovial fluid and 9.1% of PBL. These data suggest that the expression of alpha 4 beta 7 integrin may provide a mechanism whereby certain T cells adhere to rheumatoid synovium while others remain in the synovial fluid. The augmented expression of alpha 4 beta 7 in the synovial membrane T cells may contribute to the development and perpetuation of rheumatoid arthritis.

Adult↗

[Activated C3 in the synovial fluid of patients with rheumatoid arthritis and arthrosis (correlations with C3, C4 and C3 proactivator/modification caused by storage)].

Cleavage products of C3 in synovial fluid were determined by the method of 2-dimensional electrophoresis (Laurell). Statistically significant differences were observed in rheumatoid synovial fluid compared with the osteoarthrosis group. Storage is of crucial importance for the in vitro activation of C3. In fresh synovial fluid there was a significant positive correlation between breakdown products of C3 and C3, C4 and C3-proactivator. After storage (even in -70 degrees C) this positive correlation in the rheumatoid synovial fluid changes into a negative one. This phenomenon is not observed in synovial fluid from patients with osteoarthrosis. There is evidence that in the rheumatoid synovial fluid C3-activating fractions in 40S, 19S and 2,5S are responsible for the generation of C3 cleavage products.

Arthritis, Rheumatoid↗

Inhibition of Fc-rosette formation by synovial fluids from patients with rheumatoid arthritis.

Human red cells sensitized with one of the Rh antisera (Ripley) form rosettes (Fc-rosette) with human B lymphocytes and the rosettes are well inhibited by aggregated human IgG. As synovial fluids from patients with rheumatoid arthritis are usually hypocomplementaemic and frequently contain immune complexes, they were used for the inhibition of Fc-rosette formations in this study. The Fc-rosette inhibitory rates were inversely correlated with the levels of CH50 and C4 in the synovial fluids. Fractionation studies of the synovial fluids by ultracentrifugation and Sephadex G200 gel filtration indicated that main inhibitory activities were present in macromolecular fractions. Reduction and alkylation of the synovial fluids resulted in marked reductions of inhibitory activities. The tested synovial fluids did not contain anti-HLA activity as assessed by the microcytotoxicity test. The Fc-rosette inhibitory rates were not correlated with the levels of C3 and immunoglobulins, nor with the activity of rheumatoid factors in the synovial fluids. These results indicate that the Fc-rosette formations are inhibited by the immune complexes contained in the synovial fluids and it was confirmed that the Fc-rosette inhibition test is a simple and sensitive method for the detection of circulating immune complexes.

Arthritis, Rheumatoid↗