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PubMed1991

IL-1ra ELISA: reduction and alkylation of synovial fluid eliminates interference by IgM rheumatoid factors.

IL-1 and a specific receptor antagonist of IL-1, IL-1ra, may play important roles in the pathophysiology of rheumatoid arthritis and in other types of inflammatory synovitis. Measurement of IL-1ra in synovial fluids and in other body fluids may lead to a greater understanding of its possible activity as a modulator of the immune and inflammatory systems in vivo. Therefore, a modified sandwich ELISA was developed to measure IL-1ra protein concentration in synovial fluids. The antibodies used in this ELISA were polyclonal and derived from rabbits hyperimmunized with human recombinant IL-1ra. IgM rheumatoid factors within synovial fluid resulted in false elevation of determined IL-1ra by the sandwich ELISA through binding of the primary and secondary antibodies. Reduction and alkylation of synovial fluid samples before application to the ELISA plate eliminated the interference caused by greater than or equal to 2000 micrograms/ml IgM rheumatoid factor (latex agglutination titer of 1/5.120). This ELISA was specific for IL-1ra; there was no detection of IL-1 alpha, IL-1 beta, or lysozyme. The sensitivity of this ELISA was less than 200 pg/ml, making it a useful assay for the accurate measurement of synovial fluid IL-1ra protein concentration.

Alkylation↗
PubMed1984

Lipid peroxidation in rheumatoid arthritis: thiobarbituric acid-reactive material and catalytic iron salts in synovial fluid from rheumatoid patients.

Thiobarbituric acid (TBA)-reactive material is present in serum and knee joint synovial fluid from rheumatoid patients, consistent with lipid peroxidation occurring in vivo. The amount of TBA-reactive material in synovial fluid correlates with the concentration of iron salts present as determined by the bleomycin method, presumably because iron is an important catalyst of radical reactions in vivo. There appear to be significant correlations between the contents of TBA-reactive material and bleomycin-detectable iron in synovial fluid and the activity of rheumatoid arthritis as assessed with a clinical index of local inflammation and with various laboratory parameters.

Arthritis, Rheumatoid↗
PubMed1996

Plasma, urine, and synovial fluid disposition of methylprednisolone acetate and isoflupredone acetate after intra-articular administration in horses.

OBJECTIVE--To document plasma, urine, and synovial fluid disposition of 2 common intra-articularly administered steroid preparations, methylprednisolone acetate (MPA) and isoflupredone acetate (IPA). DESIGN--Descriptive investigation. SAMPLE POPULATION--100 mg of MPA or 4 mg of IPA was administered to 2 groups of 4 healthy sound radiographically normal female horses. PROCEDURE--Blood samples were collected at time 0 (before) and 2, 4, 6, 8, 10, 12, 24, 36, 48, 72, and 96 hours after administration of the designated steroid. Complete urine collection for measurement of designated steroid was accomplished by use of occluding 28-F balloon catheters. Synovial fluid samples were aseptically aspirated from the injected and contralateral uninjected tarsocrural joint at time 0 and 8, 24, 48, 240, and 672 hours after administration of the designated steroid. All samples were screened by ELISA to detect parent drug or metabolite equivalent, with a sensitivity of 2.5 ng/ml for MPA and 0.1 ng/ml for IPA. If drug was detected by ELISA in the plasma or synovial fluid, the samples were further quantified and specified, using HPLC with a lower limit of quantification (10 ng/ml). RESULTS--Between 2 and 12 hours after administration, plasma contained < 10 ng of MPA or IPA/ml (parent drug or metabolite equivalent), as intermittently detected by ELISA. Parent drug or metabolite equivalent was detected in the urine for 24 and 72 hours after injection of IPA and MPA, respectively. Synovial fluid from the contralateral joint contained no detectable MPA or IPA at any sample collection time. Median half-life for MPA, as detected by HPLC, was 10.3 hours (range, 6.1 to 10.6) in the synovial space. Median half-life for methylprednisolone, as detected by HPLC, was 10.4 (range, 9.9 to 32.1) hours. CONCLUSIONS--Both steroids appeared to be rapidly hydrolyzed to their respective ester forms, as detected by HPLC. The ELISA appeared to be a useful screening tool for detection of corticosteroids in this variety of body fluids.

Animals↗
PubMed1998

Changes in sulfation patterns of chondroitin sulfate in equine articular cartilage and synovial fluid in response to aging and osteoarthritis.

OBJECTIVES: To determine effects of aging on sulfation of chondroitin sulfate (CS) in articular cartilage and synovial fluid from normal equine middle carpal joints, and to determine whether CS compositional analysis can be used to assess alterations in proteoglycan turnover in degenerative cartilage obtained from horses with carpal osteochondral fractures. SAMPLE POPULATION: Carpal articular cartilage and synovial fluid from 44 cadavers with normal joints and from 16 Thoroughbred racehorses during routine carpal arthroscopic surgery. PROCEDURE: After papain/chondroitinase digestion of cartilage, CS disaccharides (unsulfated disaccharide delta Di0S, and monosulfated disaccharides delta Di4S and delta Di6S) were quantified by capillary zone electrophoresis. The CS was purified from synovial fluid chondroitinase digested, and analyzed. The CS nonreducing terminal residues, N-acetylgalctosamine (galNAc) or glucuronic acid adjacent to a 4-sulfated or 6-sulfated galNAc, were quantified. RESULTS: In cartilage, the delta Di6S-to-delta Di4S ratio increased with age; in degenerative cartilage, this ratio was not significantly different from the normal value. Percentage of delta Di0S decreased with age and was significantly higher in degenerative than in normal cartilage. The galNAc4S and galNAc4,6S represented > or = 96% of the terminal residues. There was a significant decrease in 6-sulfation of the terminal residues in degenerative cartilage. CONCLUSIONS: 6-Sulfation of internal and terminal CS residues increased with age. Cartilage degeneration in racehorses was accompanied by deposition of CS chains with altered sulfation patterns, in normal and diseased joints of horses > 2 years old, synovial fluid CS was not indicative of cartilage CS and may represent turnover products of a subpopulation of proteoglycan within the matrix.

Aging↗
PubMed1995

Synovial fluid L-lactic acid in acute arthritis of the adult knee joint.

Determination of synovial fluid (SF) lactic acid has been suggested to be an unspecific indicator of SF leukocytosis, and it is not recommended for differential diagnosis of bacterial arthritis. We analyzed the L-lactic acid content by enzymatic UV-method in 65 SF samples obtained from adult patients with acute knee arthritides. The concentration of L-lactic acid was not high in any SFs with intensive leukocytosis. The mean concentration of L-lactic acid was 13.5 mmol/l (95% confidence intervals 9.4; 17.6 mmol/l) in the synovial-fluid samples from culture-positive arthritis and 5.5 mmol/l (4.9; 6.2 mmol/l) in the synovial-fluid samples from culture-negative arthritis. Determination of SF L-lactic acid is an important part of the diagnostic setup for acute arthritis. Values > 9 mmol/l strongly support occurrence of bacterial arthritis and indicates an immediate onset of the treatment.

Acute Disease↗
PubMed2006

The diffusion of nimesulide gel into synovial fluid: a comparison between administration routes.

BACKGROUND: Nimesulide is available in gel formulation and applied mainly for topical pain management. However, its passage to the synovial fluid is not yet clear. The aim of this study was to evaluate if topical administered nimesulide passes into the synovial fluid and to compare its concentration with the oral nimesulide administration regimen. METHODS: Synovia and plasma nimesulide concentrations were investigated in patients after topical (Sulidin gel 1%) and oral (Mesulid tablet) drug administration. 34 adult outpatients who were scheduled to have an arthroscopic knee examination for mainly meniscal tears repair and who had knee pain during this period were enrolled in the first part of the study. One group received topical nimesulide gel to the skin of the knee whereas the second group received oral 2 x 100 mg nimesulide tablets, 4-7 days before the planned arthroscopy. Synovial fluid and plasma samples were taken simultaneously during the arthroscopy and analyzed using HPLC. In addition, an open-label pilot study was performed to investigate the efficacy and safety of 1-week administration of nimesulide gel. 63 knee osteoarthritis patients were asked to complete the WOMAC Osteoarthritis Index questionnaire before and 1 week after use of Sulidin gel applied 3 times daily. RESULTS: Synovia and plasma nimesulide concentrations were 19.7 +/- 8.6, 11.8 +/- 3.0 and 1958.8 +/- 397.5, 3631.9 +/- 799.3 ng/ ml for topical and oral administration groups, respectively. There was a significant (paired Student's t-test) improvement after 1 week nimesulide treatment in all WOMAC Osteoarthritis Index parameters measured. CONCLUSION: Nimesulide passes into the synovial fluid after topical administration and may have potential benefits in knee osteoarthritis treatment. The actual efficacy and safety of topical nimesulide gel administration should be investigated in a long-term, randomized, placebo-controlled clinical trial.

Administration, Cutaneous↗
PubMed1991

Early diagnosis of osteoarthrosis of the temporomandibular joint: correlation between arthroscopic diagnosis and keratan sulfate levels in the synovial fluid.

The role of osteoarthrosis (OA) and proteoglycan degradation in the pathogenesis of temporomandibular joint (TMJ) disorders has not been well established. The orthopaedic literature has demonstrated that proteoglycan degradation plays a significant role in the pathology of many joints. The purpose of this investigation was to determine if levels of immunoreactive keratan sulfate (an important component of cartilage proteoglycans) present in synovial fluid aspirates from TMJs correlated with arthroscopically demonstrated OA. Temporomandibular joint arthroscopy was performed on 25 joints in 20 patients and synovial fluid aspirates were obtained just prior to the insertion of arthroscopic cannulas. The results showed that synovial fluid aspirates from joints that arthroscopically demonstrated OA had significantly higher levels of keratan sulfate than synovial fluid aspirates from those joints that showed no evidence of OA (NON-OA). This study gives support to the theory that the pathogenesis of OA of the TMJ is similar to that of chondromalacia of other synovial joints. The combination of TMJ arthroscopy and synovial fluid analysis is an important model that can be used for investigation of the pathogenesis of TMJ disorders.

Adult↗
PubMed1980

Collagenase activity in human synovial fluids from joint diseases of diverse etiology.

The content of collagenase in latent as well as in apparent active form has been determined in synovial fluids from 21 patients with rheumatoid arthritis, 16 with non-rheumatoid inflammatory joint diseases, and 15 with degenerative joint disease. Collagenase activity was measured before and after activation of the latent enzyme by NaSCN treatment. Before activation, collagenase activity was present in the synovial fluids from 1 case of rheumatoid arthritis, 3 cases of degenerative joint disease, and 1 case of Behçet's syndrome. An excess of inhibitor was present in inactive synovial fluids. The incidence of collagenolytic activity markedly increased after treatment with NaSCN. When NaSCN-dependent collagenolytic activity was present, its value was of the same order of magnitude in all patients regardless of disease type. The diagnostic value of the finding of collagenase activity in synovial fluid, and the physiological meaning of the enzyme with reference to joint diseases, are discussed.

Arthritis↗
PubMed1987

Variation in synovial fluid analysis by hospital laboratories.

A survey of 42 hospitals revealed that the frequency of synovial fluid analysis was low (mean 3.9 per month). The mean was skewed upward by a higher frequency in 3 teaching hospitals, such that the overall median was 1.5 per month, with 15 hospitals studying 1 or less per month. There did not appear to be any consensus regarding what constitutes a routine synovial fluid analysis. Technical errors occurred, including dilution with acetic acid for cell counts and inappropriate use of automated cell counters. Twenty-six laboratories studied 4 "unknown" fluids for crystals. In 50 opportunities to detect each crystal type, calcium pyrophosphate was detected only 6 times, and sodium urate was detected 39 times. The white blood cell count reported for a single fluid ranged from 2,467-12,000/mm3 (mean +/- SD 6,683 +/- 1,992). The proportion of neutrophils identified varied from 0-30% (mean +/- SD 13.8 +/- 8.6). The majority of respondents believed that training and reference materials related to synovial fluid analysis should be improved.

Diagnostic Tests, Routine↗
PubMed1998

Proteoglycans in the synovial fluid of the temporomandibular joint as an indicator of changes in cartilage metabolism during primary and secondary osteoarthritis.

PURPOSE: The specific aim of this investigation was to assess differences between primary and secondary osteoarthritis (OA) of the temporomandibular joint (TMJ) using clinical evaluation and synovial fluid analysis for proteoglycans. MATERIALS AND METHODS: Arthroscopic surgery was performed on 101 TMJs from patients with significant pain or dysfunction and who had failed to respond to treatment. Joints were assessed for primary and secondary osteoarthritis. Synovial fluid aspirates were obtained and analyzed to determine the levels of keratan sulfate (KS) epitope and a novel 3B3(-) epitope by enzyme-linked immunosorbent assay (ELISA). RESULTS: Fifty-four patients and 67 joints had OA diagnosed by both clinical examination and arthroscopy. Primary OA was diagnosed in 14 joints (20%), and the remaining 53 joints were regarded as having secondary OA. No differences were detected in the levels of KS in the synovial fluid from the primary and secondary OA joints. Furthermore, the 3B3(-) epitope was not detectable in the synovial fluid aspirates of any TMJ. CONCLUSION: Secondary OA is a common disorder of the TMJ. However, there is no apparent difference in the metabolism of the joints with primary and secondary OA as assessed by proteoglycans in the synovial fluid. The apparent absence of the 3b3(-) epitope, in contrast to its presence in OA of other major synovial joints, suggests that there are some differences between the cartilage metabolism of the TMJ and these other joints during OA.

Adult↗
PubMed1993

Immuno-reactive human epidermal growth factor (h-EGF) in rheumatoid synovial fluids.

The immuno-reactive human epidermal growth factor (h-EGF) level, measured by a sensitive enzyme immunoassay, was significantly higher in synovial fluids or synovial tissue extracts from 89 patients with rheumatoid arthritis (RA) than in those from 53 patients with osteoarthritis. RA synovial immuno-reactive h-EGF was predominantly of a low molecular weight form (LMW h-EGF) on gel filtration chromatography. Furthermore, in the RA group, the synovial immuno-reactive h-EGF correlated positively with C-reactive protein, an acute-phase reactant, in the blood examination and with synovial immunoglobulin M. These results suggest that synovial h-EGF is specifically produced by RA synovium from the initial stage of arthritis, and that the measurement of synovial h-EGF serves as an early indicator to appraise the RA activity. A pathogenesis of RA including growth factors and growth inhibitory factors are discussed.

Adult↗
PubMed2002

Temporomandibular joint synovial fluid sampling: estimation of dilution factor using calcium ion concentration.

Saline aspirates have been commonly used in the biochemical investigations of temporomandibular joint (TMJ) pathology. However, due to presence of adhesions in the diseased temporomandibular joint, full equilibration between the injected saline and the synovial fluid may not be achieved in all cases. We measured calcium ion concentration in the saline aspirates and the plasma to assess the degree of dilution of the synovial fluid by the injected media. Saline aspirates obtained prior to the arthroscopic examination of 17 patients with painful TMJs not responding to 3 months of conservative treatment were analysed for their calcium content by a highly sensitive spectrophotometric autoanalyser. In 10 patients with unilateral symptoms, the contralateral asymptomatic side was used as a control. Using a concentration volume equation the amount of the synovial fluid in the saline aspirates was calculated. The yield of the saline aspirates was variable ranging from 330 to 1000 microl. The mean calcium level was 0.787 mg/dl in the symptomatic group (C.I. 95% 0.337-1.237 mg/dl) and 0.512 mg/dl (C.I. 95% 0.235-0.797) in the asymptomatic group. Using a Student t-test there was no significant difference between the two groups. Furthermore, there was no demonstrable correlation between the volume of the aspirate and its synovial fluid content. This study confirms that the saline aspirate may not be a representative sample of the TMJ synovial fluid, and that expression of the results of the biochemical assays per volume of the aspirate may be misleading.

Adult↗
PubMedDate not supplied

Effects of synovial fluid on the respiratory burst of granulocytes in rheumatoid arthritis.

Neutrophil infiltration in the synovia is an important feature of the local inflammatory process associated with rheumatoid arthritis. The present study is focused on the effects exerted in vitro by the synovial fluid versus serum on the respiratory burst of granulocytes isolated either from blood or synovial fluid of rheumatoid arthritis patients. The respiratory burst was evaluated as superoxide anion release, by lucigenin-amplified chemiluminescence. Our data show that the respiratory burst of granulocytes isolated from rheumatoid arthritis patients might trigger a significant oxidative stress both in periphery and the inflamed joint. These cells show no pathological pattern when activated in vitro by the chemotactic peptide fMLP, heterologous synovial fluid or serum. Acellular synovial fluid amplifies the superoxide anion release induced by fMLP more than the corresponding serum, indicating that a bacterial infection in the joint might enhance the oxidative damage in the inflamed synovium.

Arthritis, Rheumatoid↗
PubMed1990

Synovial fluid concentration of five different cytokines in rheumatic diseases.

Interleukin-1 beta, interleukin-2, tumour necrosis factor alpha, and the interferons, alfa and gamma, were measured concurrently in synovial fluid samples from 68 patients with rheumatic diseases. Mean interleukin-1 beta concentrations (130.3 (SD 22) pg/ml) were higher in synovial fluids from patients with rheumatoid arthritis (RA) than in those from patients with osteoarthritis (27.8(4.5)pg/ml), while measurements in synovial fluids from patients with seronegative spondarthritis were intermediate (72.7 (32) pg/ml). Interleukin-2 and tumour necrosis factor alpha concentrations were lower in the inflammatory arthropathies (RA: 4.5 (0.6) U/ml, 0.39 (0.04) ng/ml; seronegative spondarthritis: 3.1 (0.3) U/ml, 0.33 (0.03) ng/ml respectively) than those in patients with osteoarthritis (5.2 (0.6) U/ml; 0.05 (0.04) ng/ml). Interleukin-2 and tumour necrosis factor alpha concentrations correlated in all groups (r = 0.7), as did the interferons alfa and gamma (r = 0.7). There was no relation between interleukin-1 beta and either interleukin-2 or tumour necrosis factor alpha, or between the interferons and any other cytokine. Several distinct cytokine patterns were noted. Synovial fluids from two non-arthritic subjects were also examined: interleukin-1 beta concentrations were low, but concentrations of the other cytokines were higher than those seen in most arthritic fluids.

Adult↗
PubMed1991

Comparison of high-performance liquid chromatography with a radiometric assay for determination of the effect of intra-articular administration of corticosteroid and saline solution on synovial fluid hyaluronate concentration in horses.

Two recently developed direct methods, radioassay-125I-labeled hyaluronic acid binding protein (125I-HABP)- and high-performance liquid chromatography (HPLC), were used to assess and compare the concentration of hyaluronate (HA) in synovial fluid of horses. Also determined were changes in the HA concentration in an experimental treatment model involving physiologic saline solution (PSS)-irrigated or methylprednisolone acetate-injected tarsocrural joints of clinically normal horses. Serum HA concentration was determined simultaneously, using the 125I-HABP assay. Synovial fluid HA concentration values obtained by use of the HPLC method were approximately double the values obtained by use of 125I-HABP assay. Correlation (r = 0.819) between the 2 methods was highly significant (P less than 0.001; linear regression analysis) for all samples studied and for various experimental subgroups. When pure HA standards were used, correlation between the 2 methods was close to 1 (r = 0.965; P less than 0.001), with higher values obtained by use of the 125I-HABP assay. It is suggested that the HA binding protein derived from endogenous cartilage proteoglycan interferes with the 125I-HABP assay on synovial fluid, resulting in excessively low values, compared with those obtained using the HPLC procedure. Intra-articular injection of methylprednisolone acetate significantly (P less than 0.01) increased synovial fluid HA concentration at 24 hours after injection. Increase was also detected after PSS irrigation, but owing to wide intersubject variation, this increase was not significant. The HPLC procedure, which provides simultaneous information about the concentration and degree of polymerization of HA, is recommended for the study of synovial fluid, whereas the 125I-HABP assay is more suitable for serum HA analysis.

Animals↗
PubMed2005

Increased pentosidine, an advanced glycation end product, in serum and synovial fluid from patients with knee osteoarthritis and its relation with cartilage oligomeric matrix protein.

BACKGROUND: Pentosidine, an advanced glycation end product, increasingly accumulates in articular cartilage with age, and contributes to the pathogenesis of osteoarthritis (OA). Increased pentosidine concentrations are associated with inflammatory disorders-for example, rheumatoid arthritis. OBJECTIVE: To compare pentosidine serum concentrations in patients with knee OA and in healthy volunteers and to determine a relationship between pentosidine and cartilage oligomeric matrix protein (COMP)-a marker of articular cartilage destruction. METHODS: Paired serum and synovial fluid samples were obtained by arthrocentesis from 38 patients with knee OA and from 38 healthy volunteers. Pentosidine concentration was measured by reverse phase high performance liquid chromatography with fluorescent detection and COMP was determined by sandwich ELISA. RESULTS: Significantly increased serum pentosidine (p<0.01) and COMP (p<0.05) levels were detected in the patients with OA compared with the control group. Serum pentosidine correlated significantly with synovial fluid pentosidine (p<0.001). Pentosidine in synovial fluid (p<0.05) and in serum (p<0.05) correlated significantly with synovial fluid COMP. Pentosidine and COMP concentrations did not correlate significantly with the radiological stage of the disease. CONCLUSION: Increased pentosidine serum concentration in patients with OA and its correlation with the cartilage destruction marker COMP in synovial fluid suggests that pentosidine may be important in OA pathology and is a new potential OA marker.

Aged↗
PubMedDate not supplied

Synovial fluid and clinical changes after arthroscopic partial synovectomy of the equine middle carpal joint.

Changes in synovial fluid and clinical variables after arthroscopic partial synovectomy of the middle carpal joint were studied in 12 normal horses. A 7 mm motorized synovial resector was inserted into each middle carpal joint; one middle carpal joint of each horse was randomly selected to have arthroscopic synovectomy (treated) and the opposite joint was lavaged (control). Lameness examinations and synovial fluid analyses were performed before operation and at 8, 14, 21, and 28 days after operation. Lameness variables did not differ between treated and control legs. Middle carpal and carpometacarpal joint circumference measurements were increased for 4 weeks. Synovial fluid specific gravity, pH, total protein, albumin concentration, and alpha-1-, beta- and gamma-globulin concentrations, at 8 and 14 days were significantly higher than before operation in both treated and control middle carpal joints. No significant differences were found between treated and control middle carpal joints at any time for color, clarity, pH, mucin clot formation, total protein, albumin, and globulin fractions. Arthroscopic partial synovectomy and lavage did not cause significant lameness and resulted in a synovitis indistinguishable from synovitis related to arthroscopic lavage alone.

Albumins↗
PubMed1992

Activation of synovial fluid T lymphocytes by 60-kd heat-shock proteins in patients with inflammatory synovitis.

OBJECTIVE: Synovial fluid lymphocytes from patients with rheumatoid arthritis and with other forms of inflammatory synovitis demonstrate enhanced proliferative responses to Mycobacterium tuberculosis antigens, in particular, the 65-kd heat-shock protein. There is a high degree of homology between the human and the mycobacterial 60-kd family of heat-shock proteins. These studies were performed to determine if the enhanced response to the mycobacterial 65-kd heat-shock protein was due to cross-reactivity of an immune response generated against the human homolog. METHODS: These studies were performed by in vitro culture of isolated synovial fluid mononuclear cells with crude and purified antigens. RESULTS: The synovial fluid lymphocytes of a majority of patients with rheumatoid arthritis recognized the mycobacterial 65-kd heat-shock protein, as evidenced by T cell proliferation. In contrast, only 18% of all samples tested responded to a highly purified recombinant human 60-kd heat-shock protein. With only one exception, proliferative responses to the mycobacterial antigen were stronger than those to the human homolog. The proliferative responses generated against mycobacterial 65-kd heat-shock proteins from different sources were highly correlated. CONCLUSION: The findings suggest that the enhanced proliferative response to the mycobacterial 65-kd heat-shock protein noted in most patients with rheumatoid arthritis and other forms of inflammatory synovitis is not due to cross-reactivity of an immune response directed against the human heat-shock protein.

Antigens, Bacterial↗