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Activation of the neutrophil myeloperoxidase-H2O2 system by synovial fluid isolated from patients with rheumatoid arthritis.

Synovial fluid isolated from 16 patients with rheumatoid arthritis activated luminol dependent chemiluminescence in bloodstream neutrophils, and the maximal activity stimulated varied over a 50-fold range. In contrast, these same fluids only activated a much lower range (two- to threefold) of maximal rates of lucigenin dependent chemiluminescence and cytochrome c reduction, two assays which only measure oxidant secretion which is independent of myeloperoxidase. Over 95% of the luminol dependent chemiluminescence activated by all samples was inhibited by azide (indicating its dependence upon myeloperoxidase), but anti-(myeloperoxidase) IgG (which specifically inhibits only the extracellular activity of this enzyme) only inhibited the response stimulated by some samples: those fluids which activated the highest luminol dependent chemiluminescence also stimulated the greatest activity of an extracellular myeloperoxidase-H2O2 system. A clear correlation was shown to exist between the activity of myeloperoxidase already present in the fluids (after its secretion from neutrophils in situ within the rheumatoid joint) and the ability of the fluid to activate luminol dependent chemiluminescence. It is concluded, therefore, that all synovial fluid samples tested possess almost equivalent levels of a factor(s) which activated O2-/H2O2 secretion and that the variations in the measured activity of the extracellular myeloperoxidase-H2O2 system are dependent upon the level of degranulation which had occurred within the joint.

Acridines↗

Amikacin penetration into synovial fluid during treatment of septic arthritis.

The concentration of amikacin from simultaneous synovial fluid and serum samples was measured on four separate occasions in a patient treated for Serratia marcescens septic arthritis. Synovial fluid levels were between 12.5 and 24.4 micrograms/ml, with concurrent serum levels of 12.1-21.0 micrograms/ml. Parenterally administered amikacin readily distributed into synovial fluid. Failure to eradicate the patient's Serratia septic arthritis with amikacin and daily arthrocentesis may have been a result of inactivation of the antibiotic arising from acidosis occurring in the synovial fluid.

Aged↗

The penetration of ceftriaxone into synovial fluid of the inflamed joint.

Fifteen patients with acute exacerbation of rheumatoid arthritis each received a 1 g bolus intravenous injection of ceftriaxone. Serum and synovial fluid was sampled at intervals between 1 h and 24 h later and assayed for ceftriaxone. Synovial fluid leucocyte counts and albumin content were measured concomitantly. Detectable levels of ceftriaxone were found in synovial fluid and serum 24 h after injection. Synovial fluid ceftriaxone concentration ranged between 66% and 100% of the concomitant serum levels. No correlation was observed between synovial fluid ceftriaxone concentration and synovial fluid leucocyte count and albumin concentration.

Arthritis, Rheumatoid↗

The chemoattractant activity of rheumatoid synovial fluid for human lymphocytes is due to multiple cytokines.

The majority of synovial fluids from 29 rheumatoid arthritis patients were strongly attractive for normal blood lymphocytes judged by assays of polarization and collagen gel invasion. While rheumatoid synovial fluids contained IL-15, IL-8, monocyte chemotactic protein-1 (MCP-1) and macrophage inflammatory protein-1 alpha (MIP-1 alpha) at levels sufficient to attract lymphocytes, inhibition of the activity of any single cytokine using specific antibody did not abolish the activity of the fluid. However combinations of anti-cytokine antibodies used together (anti-IL-15+anti-MCP-1; anti-IL-8+anti-MCP-1 or +anti-MIP-1 alpha) inhibited most of the activity, suggesting that attraction of lymphocytes by the fluids is due to a combination of attractants. Blood lymphocytes required activation by overnight culture to respond optimally, while rheumatoid synovial tissue lymphocytes responded to synovial fluids without a requirement for a period of culture. Lymphocytes derived from rheumatoid synovial fluids were poorly responsive to locomotor stimulants. Most of the responding cells from blood mononuclear cell fractions were T lymphocytes of the CD45RO isotype. Incubation in the presence of cyclosporin A or corticosteroids inhibited the response of lymphocytes to the fluids, but the presence of non-steroidal anti-inflammatory drugs (NSAIDs) and other agents used in therapy of the patients from whom the fluids were taken had no inhibitory effect.

Adult↗

Therapeutic effects on cartilage metabolism in arthritis as measured by release of proteoglycan structures into the synovial fluid.

Proteoglycans are molecules that are degraded and released from the articular cartilage into the synovial fluid early in an arthritic process. Such released proteoglycans were quantified by an enzyme linked immunosorbent assay (ELISA). The proteoglycan content in synovial fluid from patients with various knee joint arthritides was constant in two samples withdrawn five days apart. To determine if therapeutic measures were paralleled by effects directly on the articular cartilage the patients were treated with local injections of glucocorticoids. In all patients the glucocorticoids induced a reduction of the proteoglycan content in the synovial fluid, reflecting their effects on the cartilage. In two patients with spontaneous remission of their arthritis a reduction in the proteoglycan content in the synovial fluid was also noted. The quantification of proteoglycans in synovial fluid appears to have potential as a useful tool for monitoring the effects of therapy on cartilage metabolism.

Adult↗

Stimulation of osteoclast formation by inflammatory synovial fluid.

Peri-articular bone resorption is a feature of arthritis due to crystal deposition and rheumatoid disease. Under these conditions, the synovial fluid contains numerous inflammatory cells that produce cytokines and growth factors which promote osteoclast formation. The aim of this study was to determine whether inflammatory synovial fluid stimulates the formation of osteoclasts. Synovial fluid from rheumatoid arthritis (RA), pyrophosphate arthropathy (PPA) and osteoarthritis (OA) patients was added to cultures (n=8) of human peripheral blood mononuclear cells (PBMCs) in the presence and absence of macrophage colony-stimulating factor (M-CSF) and the receptor activator of NF-kappaB ligand (RANKL). Osteoclast formation was assessed by the formation of cells positive for tartrate-resistant acid phosphatase (TRAP) and vitronectin receptor (VNR) and the extent of lacunar resorption. The addition of 10% OA, RA and PPA synovial fluid to PBMC cultures resulted in the formation of numerous multinucleated or mononuclear TRAP(+) and VNR(+) cells which were capable of lacunar resorption. In contrast to PBMC cultures incubated with OA synovial fluid, there was marked stimulation of osteoclast formation and resorption in cultures containing inflammatory RA and PPA synovial fluid which contained high levels of tumour necrosis factor alpha, a factor which is known to stimulate RANKL-induced osteoclast formation.

Acid Phosphatase↗

Analysis of synovial fluid components of hydrarthrosis in long-term hemodialysis patients.

The synovial fluid components in long-term hemodialysis patients (HD; 43 knees in 43 patients) were investigated and compared with those in patients with osteoarthritis (OA; 21 knees in 21 patients) and rheumatoid arthritis (RA; 26 knees in 26 patients). The average ages in the three groups were, respectively, 60.7 years (range, 34-79 years), 63.2 years (range, 48-88 years), and 59.7 years (range, 37-76 years). The duration of hemodialysis in the HD group averaged 14.0 years (range, 4-24 years). The concentrations of hyaluronic acid, protein, and isomers of chondroitin sulfate (chondroitin 6-sulfate [C6S] and chondroitin 4-sulfate [C4S]) in the synovial fluid, and its viscosity were measured. Differences in each of the parameters were investigated according to disease clinical stage, roentgenological grade, and periods of dialysis in the HD group. The viscosity of the synovial fluid and the concentration of hyaluronic acid in HD patients were similar to those in OA patients; however, the C6S/C4S ratio in the synovial fluid of HD patients was similar to that in RA patients. The latter finding suggests that synovitis may be present in the hydrarthrosis of HD patients. The cause of this synovitis in HD patients remains to be elucidated.

Adult↗

Joint immobilization reduces synovial fluid hyaluronan concentration and is accompanied by changes in the synovial intimal cell populations.

OBJECTIVES: Synovial fluid (SF) of normal joints contains high hyaluronan (HA) concentrations. However, the mechanism by which these are controlled and how they are influenced by articulation and loading are not established. In this study, we have examined whether immobilization influences SF HA concentration and whether this is associated with alterations in the synovial lining. METHODS: Hock joints of five adult sheep were immobilized by external fixation. Twelve weeks later, SF and synovium samples were collected. The HA concentration in SF was assayed using an ELISA-based method. Non-specific esterase (NSE) and uridine diphosphoglucose dehydrogenase (UDPGD) activities were assessed in cryostat sections of snap-chilled synovial samples using cytochemical techniques, and UDPGD activity per cell was measured in synovial lining cells by scanning and integrating microdensitometry. RESULTS: We found that the SF HA concentration was decreased from 1.65+/-0.25 mg/ml in control joints to 0.68+/-0.16 mg/ml in immobilized joints. Synovial intimal cell UDPGD activity decreased from 18.0+/-2.7 U/cell in control joints to 12.2+/-1.5 after immobilization. There was also a decrease in UDPGD-positive intimal cell numbers. Intimal surfaces in controls contained numerous NSE-positive cells, which were rarely observed in intima from immobilized joints. CONCLUSIONS: These results suggest that immobilization decreases SF HA levels and that this is associated with reduced intimal cell UDPGD activity (essential for HA formation). Immobilization also decreased the prevalence of (NSE-positive) intimal macrophages. These findings suggest that mechanosensitive homeostatic mechanisms exist within the synovial intima.

Animals↗

Gonococcal arthritis-dermatitis syndrome. Study of serum and synovial fluid immune complex levels.

Immune complexes from serum and synovial fluid were detected by the C1q binding assay in 12 patients with disseminated gonococcal infection. Since immune complexes were regularly higher in synovial fluids than in paired sera and were not detected by the monoclonal rheumatoid factor radioimmunoassay, we suggest that IgM may be present in these complexes and that this represents a primary immune response. In contrast, only 2 of 10 patients with local gonococcal infection were slightly positive both with the C1q assay and the monoclonal rheumatoid factor assay. In patients with disseminated gonococcal infection, immune complexes closely paralleled the disease activity and negatively correlated with complement levels. Synovial fluid immune complexes seem to occur in the early and aseptic phase of polyarthritis and to aid the entrance of circulating gonococcal organisms. From the results of our study, it seems that immunologic processes initiate and/or sustain inflammation in disseminated gonococcal infections that appear, at least in part, as a form of reactive arthritis.

Adolescent↗

Do synovial fluid acute phase proteins from patients with rheumatoid arthritis originate from serum?

This study was performed in order to gain insight into the occurrence, glycosylation and the possible origin of the acute-phase proteins alpha1-acid glycoprotein (AGP) and alpha1-protease inhibitor (PI) in sera and synovial fluid from patients with rheumatoid arthritis (RA). Therefore paired sera and synovial fluid samples from patients with RA, and paired synovial fluid samples from right and left knees of patients with varying degrees of arthritis were studied. Crossed affinity immunoelectrophoresis (CAIE) was used with concanavalin A and Aleuria aurantia lectin for the detection of the degree of branching and fucosylation, respectively, and the monoclonal CSLEX-1 for the detection of Sialyl Lewis(X) (SLe(X)) groups on AGP. For PI, not only CAIE, but also high-pressure-anion-exchange chromatography with pulsed amperometric detection was used to study the glycosylation. It was established that the concentrations of AGP and PI were increased in the serum of RA patients compared to normal healthy controls, but that the concentration of both proteins, as well as albumin, was significantly lower in synovial fluid than in serum. Furthermore, the type of glycosylation of both AGP and PI found in RA was significantly different from that found in normals, with increased fucosylation, but there were no major differences in the degree of branching of AGP- or PI-glycans in RA, compared to normals. No differences in glycosylation could be established between serum and synovial fluid in RA. For PI an increased fucosylation was found, both in serum and synovial fluid, using both methods of detection, and it could be established that only the alpha1-->3- and not the alpha1-->6-fucosylation of PI was affected by RA. The increased fucosylation of AGP resulted in an increased expression of SLe(X) on AGP-glycans. Since the alpha1-->3-fucosylation of AGP was significantly increased in both serum and synovial fluid from RA patients, and this correlated with systemic but not with local disease parameters, it can be suggested that acute phase proteins in synovial fluid are most probably of hepatic origin.

Adult↗

Hyaluronan molecular weight and polydispersity in some commercial intra-articular injectable preparations and in synovial fluid.

OBJECTIVE AND DESIGN: Hyaluronan is the major non-proteinaceous component of joint synovial fluid and is responsible for the unique rheological and biological properties of this medium. In joint arthropathies the molecular weight and concentration of hyaluronan may change, thereby influencing joint physiology and function. Intra-articular administrated hyaluronan derived from a number of sources, has been used for the treatment of osteoarthritis, however, there is limited information on the molecular weight and polydispersity of these various commercial preparations. The objective of this study was to develop an accurate, convenient method by which the molecular weight and polydispersity of hyaluronan may be determined and then applied to characterise the hyaluronan in synovial fluid. MATERIALS AND METHODS: Characterisation of the molecular parameters of hyaluronan of different origins and in ovine synovial fluid was accomplished using a multi-angle laser-light scattering (MALLS) detector coupled to a gel permeation chromatography (GPC) system, fitted with an automatic sample injector. CONCLUSION: Seven commercially available hyaluronan preparations of reported molecular weight were analysed. The weight average molecular weight (Mw) and number average molecular weight (Mn) values obtained for 6 of the 7 preparations using the MALLS-GPC system were in good agreement with the reported values. The abnormally low values for the exception suggested that degradation of hyaluronan had occurred. The MALLS-GPC technique was then used to determine the molecular characteristics of the endogenous hyaluronan in normal ovine synovial fluids. While the Mws ranged from less than 1 x 10(6) Da to 7 x 10(6) Da the majority were between 1-3 x 10(6) Da. [mean Mw = 2.42 x 10(6), mean Mn = 2.21 x 10(6) Da]. The effects of freezing and thawing synovial fluid upon molecular weight of hyaluronan were also investigated and were found to diminish both Mz and Mw values.

Animals↗

Abnormalities in synovial fluid of patients with septic arthritis detected by gas-liquid chromatography.

Gas liquid chromatography was performed on synovial fluid of 94 patients with exudative arthritis. A relatively constant pattern of peaks was obtained in all synovial fluid samples. Lactic acid was increased in synovial fluid of patients with septic arthritis excluding gonococcal arthritis. In addition 2 constant peaks with retention time of 546 and 848 seconds in the chromatogram usually showed a significant increase in the synovial fluids of patients with septic arthritis (including gonococcal arthritis) but not in those from patients with sterile inflammatory or degenerative arthritis. In 1 patient with pseudomonas arthritis an increase in the peaks was noted with clinical and bacteriological relapse and subsequently there was a gradual return to normal levels during clinical improvement. Increase in synovial fluid lactic acid is useful in the diagnosis of septic arthritis. Identification of the compounds represented by the 2 peaks, which presumptively correspond to n-valeric and n-hexanoic acid, may provide further information on diagnosis, prognosis and pathogenesis of arthritis and may help in the differentiation between gonococcal and nongonococcal arthritis.

Adolescent↗

Cartilage oligomeric matrix protein: a novel marker of cartilage turnover detectable in synovial fluid and blood.

Cartilage oligomeric matrix protein (COMP) is a tissue specific non-collagenous matrix protein. We have developed an enzyme-linked immunosorbent assay for the detection of this protein in synovial fluid and serum. The protein has been quantified in these fluids in patients with rheumatoid arthritis (RA), reactive arthritis, juvenile chronic arthritis, osteoarthritis and in sera of control subjects. The protein was detectable in all fluids and the synovial fluid levels were always higher than in serum in paired samples. The highest knee joint synovial fluid levels were found in reactive arthritis patients and the lowest in RA patients with advanced destruction of the knee joint. However, the relative synovial fluid content of COMP was higher in these RA patients than in patients with advanced osteoarthritis. In patients with long-standing reactive synovitis the concentrations decreased. This decrease, however, was less marked than for proteoglycan concentrations. The serum concentrations were low in patients with juvenile chronic arthritis and in patients with RA with advanced cartilage destruction of the studied knee joint. In the other groups serum levels did not differ between groups or from controls.

Adult↗

Inhibition of myeloperoxidase by synovial fluid and serum.

An inhibitor of myeloperoxidase has been identified in the synovial fluids and sera from patients with rheumatoid arthritis and sera from normal subjects. Initially, these fluids were found to inhibit stimulus induced degranulation of polymorphonuclear leucocytes independently of the stimulating agent. Subsequently, the fluids were shown to inhibit the released enzyme rather than the degranulation response of polymorphonuclear leucocytes. Both rheumatoid and normal serum samples contained high concentrations of the inhibitor but the concentrations were lower in rheumatoid synovial fluids. The inhibitory activity seemed to be specific for peroxidase as the fluids did not inhibit beta-glucuronidase activity. A protein of relative molecular mass (Mr) 150 kd was purified from synovial fluid by affinity chromatography on myeloperoxidase-Sepharose. It is concluded that serum and synovial fluid contain a novel myeloperoxidase inhibitor, which acts by binding to myeloperoxidase and thereby prevents myeloperoxidase releasing oxidative products in serum.

Adult↗

In vitro and in vivo spin echo diffusion imaging characteristics of synovial fluid: potential non-invasive differentiation of inflammatory and degenerative arthritis.

OBJECTIVE: This study was undertaken to analyse the diffusion characteristics of synovial fluid in degenerative and inflammatory arthropathies. DESIGN AND PATIENTS: Ten in vitro specimens of synovial fluid from patients with both degenerative and inflammatory arthropathy were studied at body temperature with a navigator-corrected spin echo diffusion sequence (B values 0-512 s/mm2), on a Philips 1.5-T Gyroscan. Subsequently synovial fluid from knee joint effusions of 25 patients (10 patients with osteoarthritis, 10 patients with effusions following trauma and 5 patients with effusions secondary to inflammatory arthritis) was evaluated with the same navigator-corrected spin echo diffusion sequence. RESULTS: Both in vitro and in vivo study demonstrated decreased diffusion in patients with effusions secondary to degenerative joint disease (less than 2.40 x 10(-5) cm2/s) relative to patients with effusions accompanying knee trauma (greater than 2.75 x 10(-5) cm2/s) and inflammatory arthritis (in vitro and in vivo greater than 3.00 x 10(-5) cm2/s). CONCLUSION: Synovial fluid in degenerative arthritis shows less diffusion or free water movement than synovial fluid in inflammatory arthritis. Diffusion characteristics of synovial fluid may be used to predict the nature of the underlying form of arthritis in patients presenting with knee joint effusions.

Diagnosis, Differential↗

Evidence for clonal expansion of T cell receptor V gamma II+ T cells in the synovial fluid of patients with arthritis.

We have demonstrated among synovial fluid T cells a unique profile of V gamma II sequences likely arising from clonally expanded T cells. We have determined the junctional diversity associated with each expressed V gamma family by resolving amplified fragments of cDNA into component parts on large denaturing gels. Among synovial fluid T cells we frequently find dominant fragments of a unique size clearly smaller than the dominant band observed with peripheral blood T lymphocytes. In some cases the dominant bands are 12 or 15 nucleotides smaller than the corresponding most abundant band from peripheral blood T lymphocytes. Patterns of lower m.w. species not typical of a polyclonal population argues that clones of T cells expressing the V gamma II family are expanding in the joint and that a high proportion of these cells do not express the V gamma IIJP sequence typical of peripheral blood but rather express V gamma II in combination with a shorter J fragment, JP1, JP2, J1, or J2. In addition by examining joint effusions from the left and right knees from the same individual we have shown that the profiles of V gamma II sequences derived from the fluids are identical to each other but clearly distinct from that of peripheral blood. We have, in addition, quantitated with a series of synthetic internal standards the relative usage of each V gamma family expressed by T cells in the synovial fluid and peripheral blood of seven patients with arthritis including six patients who were either children or adolescents and one adult patient. All patients showed a reduction in the relative expression of V gamma II in synovial T cells relative to peripheral blood T lymphocytes and a corresponding increase in the expression of V gamma I or V gamma III or both. We did not detect expression of V gamma IV in either lymphocyte population.

Arthritis↗

The sulphated glycosaminoglycan levels in synovial fluid aspirates in patients with acute and chronic joint disease.

Proteoglycan levels were measured in a series of synovial fluid samples from patients with acute and chronic joint diseases using a modified chemical dye binding method. Levels found in 50 miscellaneous inflammatory arthritis fluids (mean = 173.2 +/- 90.9 micrograms/ml) were higher than found in either 50 with rheumatoid arthritis (96.3 +/- 31.3 micrograms/ml) or 50 with osteoarthritis (83.8 +/- 27.3 micrograms/ml). For comparison, proteoglycan levels were measured in 15 cadaver synovial fluids (98.9 +/- 44.2 micrograms/ml) and 12 synovial fluids from patients with sports injury (163.7 +/- 79.4 micrograms/ml). Patients were recruited into a trial where synovial fluid was aspirated as often as possible over a 6-month period during which the patients were followed using a number of well proven clinical parameters. No correlation was found between the degree of joint destruction as measured by X-ray damage and the concentration or total amount of proteoglycan in the synovial fluid. In addition, there was no correlation between the level or total amount of proteoglycan and any clinical parameter of disease activity.

Acute Disease↗

Complement activation in seropositive and seronegative rheumatoid arthritis. 125I-C1q binding capacity and complement breakdown products in serum and synovial fluid.

1. The detection and quantitation of immune complex-like material in synovial fluid and in serum from patients with joint diseases was done through the measurement of the capacity to bind radiolabeled C1q. It was found that 65% of synovial fluid samples from seropositive or seronegative RA patients had a high C1q binding capacity as compared to other joint diseases. Immune complex-like material was also detected in 63% of serum samples from seropositive RA patients. 2. The existence of C3 or C3PA breakdown products in synovial fluid from most of the synovial fluids from RA patients probably reflects an activation of the complement system occurring in both forms of the disease. C3PA breakdown products were never found in degenerative or post-traumatic joint diseases and only occasionally in other inflammatory arthritis. Apart from their pathogenic significance, these results may have some interest for the clinical investigation of patients with joint diseases.

Arthritis, Rheumatoid↗