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Cartilage macromolecules in knee joint synovial fluid. Markers of the disease course in patients with acute oligoarthritis.

OBJECTIVE: To investigate the development of chronic joint symptoms in patients presenting with acute oligoarthritis including knee joint synovitis with effusion and explore whether prognostic information can be derived from initial synovial fluid concentrations of aggrecan and cartilage oligomeric matrix protein (COMP) for development of chronic joint symptoms. METHODS: Retrospective follow up of 25 patients identified in a bank of knee joint synovial fluids collected consecutively from patients presenting with knee joint synovitis and symptoms from at most three additional joints and in whom no diagnosis could be established at presentation. RESULTS: The 10 patients who developed chronic joint symptoms were characterised by lower knee joint synovial fluid concentrations of aggrecan as well as lower aggrecan/COMP ratios (p < 0.001) than the 15 patients who had a transient arthritis. No other clinical or laboratory differences between the groups were apparent at the time of presentation. CONCLUSIONS: The synovial fluid content of aggrecan is a potential tool in acute arthritis for distinguishing patients with a benign disease course from those who will develop a chronic joint disorder.

Acute Disease↗

Vascular endothelial growth factor concentrations in synovial fluids of patients with symptomatic internal derangement of the temporomandibular joint.

BACKGROUND: Vascular endothelial growth factor (VEGF) is an inducer of angiogenesis and permeability of small blood vessels. We determined the concentrations of VEGF in synovial fluid of patients with symptomatic internal derangement of the temporomandibular joint (TMJ). METHODS: Diluted synovial fluid was collected by a pumping procedure from 22 TMJs of patients with internal derangement and 10 control TMJs. VEGF concentration was determined by an enzyme-linked immunosorbent assay. RESULTS: The VEGF was detected in 14 of the 22 joints (64%) of patients with internal derangement, at a mean concentration of 67 pg/ml, but in only one control joint (12.5 pg/ml) (P = 0.004 for the difference in concentration). There was a significant correlation between VEGF concentration and total protein concentration in the synovial fluid (P = 0.002). CONCLUSIONS: The increased concentration of VEGF in patients with symptomatic internal derangement suggests that this growth factor may be involved in the pathogenesis of this condition.

Adolescent↗

Regulatory effects of IL-13 on synovial fluid macrophages and blood monocytes from patients with inflammatory arthritis.

Activated macrophages are central to the destructive processes of chronic inflammatory arthritis. In this study, it was hypothesized that IL-13, a product predominantly of 'Th2-type' lymphocytes, may be used therapeutically to down-regulate monocyte/macrophage activities at sites of chronic inflammation. Synovial fluid mononuclear cells were isolated from 12 patients with chronic inflammatory arthritis. Peripheral blood mononuclear cells (PBMC) were isolated at the same time as synovial fluid cells from all 12 patients. IL-13 significantly inhibited lipopolysaccharide (LPS)-induced tumour necrosis factor-alpha (TNF-alpha) production by mononuclear cells from peripheral blood, but not synovial fluid. In contrast, IL-13 inhibited LPS-induced IL-1 beta production by all cells, and as a positive response to IL-13, CD23 expression was increased on both cell populations. Blood monocytes cultured for 7 days with granulocyte-macrophage colony-stimulating factor (GM-CSF) or M-CSF responded to IL-13 in a manner similar to that detected for synovial fluid-derived cells, with suppression of LPS-induced IL-1 beta, but not TNF-alpha, production. In all experiments, the responses to IL-13 were very similar to those detected to IL-4, but differed from those measured with IL-10. Thus, the responses to IL-13 by synovial fluid cells and cultured monocytes are not equal to those of blood monocytes. The similar responses to IL-4 and IL-13 support claims of a common element for signalling from the IL-4 and IL-13 receptors. Furthermore, the activity of a common receptor chain may be altered by monocyte activation and differentiation.

Antigens, CD↗

Determination of cytidine deaminase activity in synovial fluid by HPLC.

A reversed-phase high-performance liquid chromatographic method for the determination of cytidine deaminase activity in synovial fluid is described. Diluted synovial fluid was incubated for 10 min at 56 degrees C with 0.4 mM cytidine. The protein in 5 vol of incubate was then precipitated using 1 vol of trichloroacetic acid (20% m/v) and the substrate, cytidine, and the product, uridine, were determined in the resultant supernatant. These substances were separated by reversed-phase HPLC using 0.05 M potassium dihydrogen orthophosphate (pH 6.5) containing methanol (3% v/v) and were detected at 280 nm. The enzyme activity was determined by measuring uridine formation. The effects of substrate concentration, pH and reaction temperature on uridine formation are described.

Ammonia↗

Normal human synovial fluid and articular cartilage contain similar intact proteoglycans.

This work, directed to characterization of proteoglycans present in normal human synovial fluid by Western blotting techniques, revealed an intimate relationship of these proteoglycans to those of articular cartilage. Analyses were performed on samples subjected to digestion with chondroitinase ABC, in the presence or absence of keratanase, yielding products containing core proteins with glycosaminoglycan side chain stubs. The proteoglycan core proteins contained epitopes reactive with monoclonal antibodies that distinguish between chondroitin sulfate-4 and chondroitin sulfate-6. Additionally, these products reacted with monoclonal antibody to keratan sulfate when keratanase was omitted from the digestion. The analysis of synovial fluid revealed that the proteoglycan core proteins expressed predominantly the chondroitin sulfate-6 epitope, with expression of the chondroitin sulfate-4 epitope demonstrable only in prepubertal individuals. There was coexpression of both chondroitin sulfate epitopes in all proteoglycan core proteins of prepubertal individuals. Coexpression of chondroitin sulfate and keratan sulfate epitopes occurred in all proteoglycan core proteins. Proteoglycan core proteins had M(r) similar to those obtained from articular cartilage. Hence, in individuals free of joint disease, most proteoglycans seem to be transferred from articular cartilage to the synovial fluid without major alteration in the apparent size of the proteoglycan core protein. Only a minor set of proteoglycan core proteins had no direct articular cartilage equivalent. As this set also contained keratan sulfate, it is likely to be of articular cartilage origin, but probably modified by proteolysis.

Adolescent↗

Leukocyte count in the synovial fluid of children with culture-proven brucellar arthritis.

Brucellosis is an important cause of paediatric septic arthritis in endemic areas. Because the Gram stain is frequently negative and culture results are unavailable at the time of the patient's admission, the diagnosis of brucellar arthritis is usually entertained on the bases of epidemiological considerations and cytological examination of the synovial fluid aspirate. The aim of this study was to assess the sensitivity of a synovial fluid leukocyte count >50 000 WBC/mm(3) for detecting culture-proven brucellar arthritis in children. The medical records of all children with brucellar arthritis diagnosed since 1994 in a hospital serving an endemic area for brucellosis in southern Israel were reviewed. Nine patients (six males and three females), aged 3-14 years, were identified. A single joint was affected in all patients. The median leukocyte count in the synovial fluid was 9500 WBC/mm(3) (range 300-61 500 WBC/mm(3)), and in eight of the nine patients it was less than 50 000 WBC/mm(3). Brucella melitensis was recovered from the synovial fluid culture in all patients. The diagnosis of brucellar septic arthritis cannot be excluded on the basis of a low leukocyte count in the joint aspirate. A high index of suspicion and use of modern culture techniques are recommended to improve the diagnosis of brucellar arthritis.

Adolescent↗

Immune complexes in sera and synovial fluids of patients with rheumatoid arthritis. Radioimmunoassay with monocylonal rheumatoid factor.

Evidence for the presence of immune complexes in blood, synovial fluid, and tisues of patients with rheumatoid arthritis (RA) includes low complement levels in blood and effusions, deposition of immunoreactants in tissues and vessel walls, precipitate formation after addition of monoclonal rheumatoid factor (mRF) to serum or synovial fluid. To quantitate immune complex-like material in RA patients, we developed a radioimmunoassay based on inhibition by test samples of the interaction of (125I)aggregated IgG (agg IgG) and mRF coupled to cellulose. This method could measure immune complexes of human antibody with hemocyanine prepared in vitro. The assay was not influenced by presence of polyclonal RF in test samples, nor by freezing and thawing. Normal levels of immune complex-like material in serum were less than 25 mug agg IgG EQ/ML. 12 of 51 RA sera examined (26%) contained more than 25 mug/ml. The presence of this material in RA sera was found to correlate with severity of disease, as measured by anatomical stage and functional class. There was an inverse correlation of the material with serum C4 level. Rheumatoid synovial fluids generally contained higher levels than serum, and five of 23 contained very much higher levels. The frequency of elevated levels of immune complex-like material in sera of patients with systemic lupus erythematosus (2 of 29) and with miscellaneous vasculitides (2 of 21 was much lower than in RA, suggesting that mRF exhibits a specificity for only certain kinds of immune complexes. The reason for this apparent specificity may explain such distinctive features of RA as the high frequency of polyclonal RF, the lack of immune complex nephritis, and the generally normal levels of serum complement.

Adult↗

Immunological characteristics of lymphocytes in synovial fluid and peripheral blood in patients with rheumatoid arthritis.

The investigation has been carried out on 30 patients with rheumatoid arthritis. Rosettes (E, EA, EAC) technique and immunofluorescence test were used to detect the surface markers of lymphocytes. Cytotoxic activity of T and K cells in vitro was established. It has been proved that cells occurred to a larger extend in synovial fluid than in the peripheral blood. B cells and lymphocytes with Fc receptors were only a few in synovial fluid. The percentage ofFc receptors bearing cells was higher in the peripheral blood of patients with rheumatoid arthritis than that in peripheral blood of the control group. Cytotoxic activity of K-cell was low in synovial fluid. PHA-cell mediated cytotoxicity was low in peripheral blood as well as in synovial fluid of patients with rheumatoid arthritis.

Antibody-Dependent Cell Cytotoxicity↗

[Artificial synovial fluid for the joints].

The author presents the results of his biomechanical, experimental and clinical studies to produce an artificial synovial fluid for therapeutic correction when administered intraarticularly in inflammatory and degenerative diseases, traumas and operations on the joints. According to the results of the rheologic investigations, the 15% aqueous solution of mean molecular polyvinylpyrrolidone (PVP) and its complex with a biopolymer (PVP-hyaluronate) turned out to be most similar to the natural synovial fluid of the joints. The experimental studies (201 rabbits) revealed a suppression of the inflammatory process in the joints and an antiadhesive action of the artificial synovia which improved the metabolism of the acetabular cartilage and the functions of the joints. In the clinical part of the study (514 patients) the preparations of the artificial synovial fluid and its complexes with other drugs were administered intraarticularly to the patients with rheumatoid arthritis and deforming osteoarthrosis. An improvement in many values of the function of the joints and of the laboratory and biochemical data has been revealed. There have been no side effects in the patients under study; an improvement in the working ability has been noted, including the evaluation of the long-term results (1 to 10 years).

Animals↗

Synthesis of the active metabolite of vitamin D, 1,25(OH)2D3, by synovial fluid macrophages in arthritic diseases.

Synthesis of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) has been shown in cells from knee joint synovial fluid of 20 patients with inflammatory rheumatoid disease, reactive or psoriatic arthritis, or gout, all of which had high synovial fluid cell counts, and by cells from a patient with aseptic necrosis of a femoral condyle after short term (less than 24 hours) or long term (seven days) primary culture. Cells from 18 patients with inflammatory arthritis, five of which had low synovial fluid cell counts and cells from six patients with osteoarthritis were unable to synthesise this metabolite from 25-hydroxyvitamin D3 (25(OH)D3). Macrophages are believed to be the cells responsible for synthesising 1,25(OH)2D3 because these were significantly more numerous in samples that formed 1,25(OH)2D3; they were also the predominant cell type present in the aseptic necrosis sample and the only cell type present in preparations maintained for one week in monolayer culture.

Arthritis↗

Biochemical markers in synovial fluid identify early osteoarthritis of the glenohumeral joint.

The objective of this study on the glenohumeral joint was to assess the (1) accuracy of clinical diagnosis of osteoarthritis compared with arthroscopic diagnosis, and (2) the ability of biochemical markers in synovial fluid to detect osteoarthritis. Patients (96) were examined clinically and the preoperative diagnosis of osteoarthritis was recorded. At surgery (arthroscopy or arthroplasty), the glenohumeral joint was inspected for signs of osteoarthritis, and the joint osteoarthritis grade (I-IV) was recorded. At surgery, synovial fluid lavage was obtained from the joint, and later analyzed to determine levels of aggrecan components: total sulfated glycosaminoglycan and keratan sulfate epitope, link protein and the chondroitin sulfate epitope recognized by antibody 3B3 (3B3(-)). Compared with arthroscopic diagnosis of osteoarthritis, the results showed that the clinical diagnosis did not wrongly identify joints without osteoarthritis, and was always able to identify joints with advanced (Grade IV) osteoarthritis. Grade II osteoarthritis was rarely identified (10% of the time), and Grade III osteoarthritis was identified 50% of the time. Biochemical assessment of the synovial fluid showed that the catabolic markers (sulfated glycosaminoglycan, keratan sulfate and link protein) were elevated in fluids from joints with moderate (Grade III) and advanced osteoarthritis (Grade IV), and the 3B3(-) epitope was elevated in Grades II, III, and IV. These results show that arthroscopic diagnosis for osteoarthritis, of the glenohumeral joint is particularly useful for early and moderate osteoarthritis, where clinical (nonarthroscopic) diagnosis is poor, and that biochemical analysis of the synovial fluids corresponds well to arthroscopic diagnosis of shoulder osteoarthritis.

Arthroscopy↗

Measuring synovial fluid viscosity with a white blood cell diluting pipette. A simple, rapid,and reproducible method.

A convenient and accurate way to measure the relative viscosity of synovial fluid is to use a white blood cell diluting pipette as a simple viscometer. The technique is reproducible with 2% accuracy and adds little to the time required for routine synovial fluid analysis. The magnitude of the variation of synovial fluid viscosity with temperature is demonstrated, and it is suggested that routine clinical measurements be made at room temperature. Clinical data confirm that a single viscosity measurement has little or no diagnostic value.

Hematology↗

Comparison of serial synovial fluid cytology in rheumatoid arthritis: delineation of subgroups with prognostic implications.

Serial synovial fluid samples were obtained from patients with rheumatoid arthritis and analysed cytologically. Positive correlations in the serial samples between proportions of polymorphs and ragocytes were found. Based upon the serial synovial fluid cytology, distinct subgroups of rheumatoid joints emerged. Comparison of clinical findings in these subgroups showed that the joints with high ragocyte proportion had a significantly poorer outcome. This study therefore identifies two distinct subgroups of rheumatoid joints on the basis of the nature of the cellular reaction within the joint fluid, which usually holds true during subsequent clinical relapse. Identification of these groups may have prognostic significance.

Adolescent↗

Clonal heterogeneity of synovial fluid T lymphocytes in inflammatory synovitis.

Several studies have examined patients with rheumatoid arthritis for the presence of oligoclonal populations of synovial T lymphocytes. The results of these studies have been conflicting. In this study one patient with rheumatoid arthritis and two with other forms of inflammatory synovitis were examined by Southern blot analysis of T cell clones generated from synovial fluid by primary limiting dilution. Evidence of oligoclonality was documented only in a patient with psoriatic arthritis. The distinguishing characteristics of this patient, in addition to the diagnosis, included the fact that only one joint was involved, the synovitis in the affected joint was of recent onset, and the synovial fluid lymphocytes from which the T cells were cloned responded strongly to soluble antigens. Because of the association with the strong response to soluble antigens, synovial fluid T lymphocytes from another patient with rheumatoid arthritis were cloned in response to a crude mycobacterial antigenic mixture. Three of the seven clones examined were identical by Southern blot analysis. These observations suggest that the presence of oligoclonality is limited in patients with inflammatory arthritis. The relationship of a specific antigen-driven response within the joint to the detection of oligoclonal T cells within that joint remains to be determined.

Adult↗

The entrapment of corrosion products from CoCr implant alloys in the deposits of calcium phosphate: a comparison of serum, synovial fluid, albumin, EDTA, and water.

Physical wear of orthopedic implants is inevitable. CoCr alloy samples, typically used in joint reconstruction, corrode rapidly after removal of the protective oxide layer. The behavior of CoCr pellets immersed in human serum, foetal bovine serum (FBS), synovial fluid, albumin in phosphate-buffered saline (PBS), EDTA in PBS, and water were studied using X-ray Photoelectron Spectroscopy (XPS) and Time-of-Flight Secondary Ion Mass Spectroscopy (ToF-SIMS). The difference in the corrosive nature of human serum, water, albumin in PBS and synovial fluid after 5 days of immersion was highlighted by the oxide layer, which was respectively 15, 3.5, 1.5, and 1.5 nm thick. The thickness of an additional calcium phosphate deposit from human serum and synovial fluid was 40 and 2 nm, respectively. Co and Cr ions migrated from the bulk metal surface and were trapped in this deposit by the phosphate anion. This may account for the composition of wear debris from CoCr orthopedic implants, which is known to consist predominantly of hydroxy-phosphate compounds. Known components of synovial fluid including proteoglycans, pyrophosphates, phospholipids, lubricin, and superficial zone protein (SZP), have been identified as possible causes for the lack of significant calcium phosphate deposition in this environment. Circulation of these compounds around the whole implant may inhibit calcium phosphate deposition.

Albumins↗

Differential metabolism of 25-hydroxyvitamin D3 by cultured synovial fluid macrophages and fibroblast-like cells from patients with arthritis.

Differential metabolism of 25-hydroxyvitamin D3 (25(OH)D3) has been shown for macrophages and fibroblast-like cells (possibly synoviocytes) cultured for two to 50 days after isolation from the synovial fluid of 12 patients with various forms of inflammatory arthritis. Macrophages synthesised the active metabolite of vitamin D3, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), the synthesis of which was increased by bacterial lipopolysaccharide, a known macrophage activating factor. In contrast, fibroblast-like cells formed 24, 25-dihydroxyvitamin D3 (24,25(OH)2D3), synthesis of which was stimulated by 1,25(OH)2D3 and inhibited by lipopolysaccharide. The synthesis of 1,25(OH)2D3 and 24,25(OH)2D3 by macrophages and fibroblast-like cells respectively was inhibited by ketoconazole, indicating that both hydroxylases are dependent on cytochrome P-450. Mean (SEM) synovial fluid and serum 25(OH)D3 concentrations were 16.7 (1.7) and 22.2 (2.6) ng/ml and those of 1,25(OH)2D3 were 29.4 (4.8) and 43.3 (4.0) pg/ml respectively. In most cases concentrations were lower in synovial fluid than in paired serum samples, but in two patients 1,25(OH)2D3 concentrations were greater in synovial fluid than in serum, suggesting local synthesis within the affected joints.

24,25-Dihydroxyvitamin D 3↗

Recombinant equine growth hormone administration: effects on synovial fluid biomarkers and cartilage metabolism in horses.

REASONS FOR PERFORMING STUDY: Recombinant equine growth hormone (reGH) has recently been evaluated for effects on body condition and wound healing. It has the potential to influence articular cartilage via stimulation of IGF-1. OBJECTIVES: To investigate effects of administration on synovial joint metabolism. METHODS: Six mature horses were given 20 microg/kg bwt reGH daily for 8 weeks by i.m. injection. Three control horses were injected with sterile water. Serum and synovial fluid samples were collected at 6, 8, 11 and 16 weeks for GH and IGF-1 assays. Articular cartilage harvested at week 16 was evaluated by Western analysis using monoclonal antibodies BC-13, BC-4, 8-A-4 and CH-3. RESULTS: Concentrations of IGF-1 in serum and synovial fluid were significantly elevated (P < 0.05) at 6 and 8 weeks in the reGH group. Glycosaminoglycan concentrations in synovial fluid were significantly less than controls at these time points, suggesting that reGH may modulate proteoglycan metabolism in articular cartilage. In the reGH group, there were not any alterations in synovial fluid content of 3B3(-) epitope or aggrecan metabolite, or in aggrecan or link protein catabolites retained within cartilage, that might be expected with development of osteoarthritis. CONCLUSIONS: Intramuscular administration of reGH may be a more efficient means of delivery of IGF-1 to joints for cartilage resurfacing initiatives. POTENTIAL RELEVANCE: We found no alterations in cartilage metabolism indicative of development of osteoarthritis.

Animals↗