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Hyaluronate and large molecular weight proteoglycans in synovial fluid from horses with various arthritides.

OBJECTIVE: To investigate the presence of large molecular weight (MW) proteoglycans (PG) and hyaluronate (HA) in synovial fluid (SF) from horses with various arthritides and from control joints. DESIGN: Horses with acute (< 2 weeks) or chronic (> 4 weeks) lameness were examined by clinical examination, intrasynovial anesthesia, radiography, arthroscopy, and SF analysis. Joints were grouped on the basis of diagnosis: acute traumatic arthritis, chronic traumatic arthritis (with a subgroup of degenerative joint disease), intra-articular fracture, and infectious arthritis. ANIMALS: 31 horses with arthritis and 9 control horses; altogether 43 SF samples were analyzed. PROCEDURE: High-performance liquid chromatography was used to assess HA and large MW PG in SF samples. RESULTS: A PG peak was identified in 8 of 23 SF samples of joints with chronic traumatic arthritis, 4 of which had no or minimal abnormal radiographic findings but mild articular cartilage fibrillation detected by arthroscopy, and in 3 joints with intra-articular fracture and 1 with resolving infectious arthritis, but not in joints with acute traumatic arthritis or in control joints. There was significant difference (P < 0.01) in mean (+/- SEM) HA concentration between control joints and joints with chronic traumatic arthritis (0.32 +/- 0.04 g/L; n = 9 vs 0.18 +/- 0.01 g/L; n = 23). CONCLUSION: Large MW PG fragments are released into equine SF in the course of articular disease and can be detected simultaneously with HA by high-performance liquid chromatography. CLINICAL RELEVANCE: The SF HA concentration can be used as diagnostic marker for chronic traumatic arthritis. However, SF PG or other marker cannot be used for diagnosing or monitoring degenerative joint disease.

Animals↗

[Usefulness of anti-cyclic citrullinate peptide antibody determination in synovial fluid analysis of patients with rheumatoid arthritis].

OBJECTIVE: To assess the role of anti-cyclic citrullinated peptide (CCP) antibody detection in synovial fluid (SF) of RA patients compared to OA patients. METHODS: We evaluated in 25 RA subjects and 14 OA patients, presenting a knee-joint effusion, the main clinical and laboratory parameters including the number of painful and/or swollen joints, Ritchie index, morning stiffness, ESR, CRP and analysis of SF obtained by therapeutic arthrocentesis. IgG anti-CCP (ELISA), rheumatoid factor (RF) and total IgG (nephelometry method) were measured in SF and paired serum samples. RESULTS: We found anti-CCP antibodies and RF in 64% (16/25) and 60% (15/25) of RA sera, respectively; 72% (18/25) of RA patients were positive for anti-CCP antibodies or RF. We found a higher SF/serum ratio for anti-CCP (p<0.004) compared to that for total IgG. The calculation of anti-CCP concentration as IgG anti-CCP (units)/total IgG (g L-1) revealed higher values in SF than in serum (p<0.046) in RA patients. Among these, correlation analysis showed that anti-CCP/total IgG values in SF correlated with the relative concentration of serum anti-CCP/total IgG (rs=0.842; p<0.00001) and serum anti-CCP antibody levels (rs=0.799; p<0.0001). We did not find any correlation between SF anti-CCP levels and the main characteristics of SF as well as the clinical or laboratory parameters. CONCLUSION: Our study give evidence for a preferential production of anti-CCP antibodies at RA joint level, confirming the pathogenetic role of these autoantibodies. Moreover, SF determination of anti-CCP, corrected for the total amount of the corresponding immunoglobulin, may be helpful as diagnostic tool in selected cases.

Adult↗

Pharmacokinetic studies of methotrexate in plasma and synovial fluid following i.v. bolus and topical routes of administration in dogs.

PURPOSE: The pharmacokinetic properties of methotrexate (MTX) in the plasma and synovial fluid (SF) after bolus i.v. and topical administration were studied in dogs to assess the feasibility of topical delivery of MTX for the treatment of rheumatoid arthritis. METHODS: A MTX gel in Poloxamer 407 containing an absorption enhancer was formulated and topically applied on the elbow and stifle joints of dogs. SF was collected by inserting a needle with syringe into the joint space. Drug concentrations in the plasma, SF and muscle tissues were determined using a HPLC method with fluorimetric detection. RESULTS: Peak MTX concentrations in SF occurred at 38 +/- 5 min following bolus i.v. dose, indicating the presence of a substantial diffusion barrier between the plasma and SF. The plasma/SF concentration ratios of 1.16 +/- 0.25 were maintained after the attainment of distribution equilibrium between the two compartments. The t1/2 values in the plasma (11.2 +/- 1.2 hr) and SF (12.7 +/- 3.7 hr) were similar during the elimination phase, while the MRT in SF (3.24 +/- 0.21 hr) was longer than that in plasma (2.56 +/- 0.20 hr), probably due to the slow distribution of MTX to SF. After topical dose, MTX concentrations in plasma reached the steady state at approximately 4 hr, lasting for approximately 20 hr. The bioavailability of MTX from the gel was 11.8 +/- 3.3% of the applied dose, but muscle tissues beneath the gel application site had significantly higher levels of MTX than untreated muscle tissues. There was no statistical difference in SF concentrations of MTX between drug treated and untreated joints 24 hr after topical dose. CONCLUSIONS: Topical delivery of MTX in a hydrophilic gel achieved a sustained C/t profile in plasma and higher drug levels in muscle tissues underneath the dosing site, implicating the potential therapeutic value of the topical formulation.

Administration, Topical↗

Identification of prostaglandin E2 and leukotriene B4 in the synovial fluid of painful, dysfunctional temporomandibular joints.

It has been hypothesized that prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) should be present in the synovial fluid of inflamed, dysfunctional temporomandibular joints. An assay to identify PGE2 and LTB4 and platelet-activating factor (PAF) was used, and a strong correlation between the levels of these lipid mediators of pain and inflammation and an index of clinical joint pathology was found.

Cartilage, Articular↗

Demonstration of CD13/aminopeptidase N on synovial fluid T cells from patients with different forms of joint effusions.

Cytofluorometric analysis was performed to characterize the immunophenotype of lymphocytes of the synovial fluid (SF) and the peripheral blood (PB) from patients suffering from juvenile chronic arthritis (JCA) or rheumatoid arthritis (RA). The most obvious difference could be found in expression of the surface protease aminopeptidase N (AP N/CD13). Whereas monoclonal antibodies specific to CD13 failed to reveal surface expression on lymphocytes of the PB; 63 +/- 15% of SF T cells gave positive staining for CD13 using Leu-M7. No correlation between CD13 expression and joint disease could be found in patients who had different types of inflammatory joint effusions. CD13 expression of T cells was also found in synovial tissue and inflammatory serous cavity effusions. Fixation of T cells revealed the presence of intracellular CD13 antigen already located in the PB T cells of healthy individuals. Induction of CD13 expression on PB T cells could be demonstrated after incubation with Con A/IL-2 or SF from patients with RA. Our findings suggest a role for AP N as a new activation-associated molecule of T lymphocytes.

Aminopeptidases↗

[Alpha-2 macroglobulin in serum and synovial fluid of patients with rheumatoid arthritis and osteoarthrosis].

AIM: To study concentrations of alpha-2-macroglobulin (A-2-MG) in blood serum and synovial fluid (SF) in patients with rheumatoid arthritis and formulation of basic clinical, laboratory and x-ray criteria of RA activity. MATERIALS AND METHODS: A-2-MG levels were measured in the serum and SF from 151 RA patients and in the serum of 20 patients with osteoarthrosis (OA) and 62 healthy donors. The serum concentration for RA patients was 166 +/- 65.3 mg%, for OA patients--175.26 +/- 36.99 mg% and for healthy donors--177.772 +/- 50 mg%. Mean concentration of SF A-2-MG in RA patients was 98.77 +/- 82.43 mg%. CONCLUSION: Changes in the concentration of A-2-MG are unrelated to inflammation activity in the joints of RA and OA patients. Serum and synovial concentration of this protein corresponds to changes in the concentration of IgM and rheumatoid factors in RA patients.

Arthritis, Rheumatoid↗

Significance of cathepsin B accumulation in synovial fluid of rheumatoid arthritis.

We measured and compared the activities of various kinds of proteinases, such as cysteine, serine, aspartic, and metalloproteinases, in synovial fluids of 16 patients with rheumatoid arthritis (RA) and 18 patients with osteoarthritis (OA). More than 19-fold higher activity of cathepsin B and about 6-fold higher activity of prolylendopeptidase, compared to those of OA, were accumulated in RA fluid. Moreover, levels of cathepsins B and S using the corresponding sandwich enzyme immunoassays were statistically higher in RA fluid than those in OA. Significant amounts of 41-kDa and 35-kDa procathepsin L were detected in RA fluid using gelatin zymography, while 41-kDa enzyme alone was detected in OA. Cathepsin B in RA fluid could degrade collagen, and this degradation was suppressed by the addition of CA-074, a specific inhibitor of cathepsin B. Therefore, cathepsin B may participate in joint destruction of RA, and its inhibitor may be effective for RA care.

Arthritis, Rheumatoid↗

Bleomycin-detectable iron in knee-joint synovial fluid from arthritic patients and its relationship to the extracellular antioxidant activities of caeruloplasmin, transferrin and lactoferrin.

Some 40% of knee-joint synovial fluids from arthritic patients show the presence of bleomycin-detectable iron. This is released from a protein component of the fluid to bleomycin at acidic pH values. Patients whose fluids release iron have lower contents of transferrin, lactoferrin and caeruloplasmin than do patients whose fluids do not release iron to bleomycin. These proteins are important extracellular antioxidants, and measured antioxidant activities are extremely low in the iron-releasing fluids. The propensity of some fluids to release iron at low pH values, characteristic of the microenvironment beneath adherent macrophages, coupled with their decreased antioxidant protection against iron-stimulated oxygen-radical damage, might explain previously reported correlations between clinical disease severity, lipid peroxide content and the presence of bleomycin-detectable iron [Rowley, Gutteridge, Blake, Farr & Halliwell (1984) Clin. Sci. 66, 691-695].

2,2'-Dipyridyl↗

Plasma and synovial fluid concentrations of salicylic acid and its metabolites in patients with joint effusions.

Equilibration of salicylic, salicyluric and gentisic acids between plasma and synovial fluid (SF) was measured in 36 patients receiving chronic salicylate therapy and from whom SF was required for diagnostic purposes. Gentisic and salicyluric acids equilibrated completely, while SF salicylic acid concentration was less than that in plasma. The presence of significant gentisic acid concentrations in SF could contribute to the therapeutic response to chronic salicylate therapy, since its antiinflammatory effect is even greater than that of acetylsalicylic acid.

Adult↗

Different ELR (+) angiogenic CXC chemokine profiles in synovial fluid of patients with Behçet's disease, familial Mediterranean fever, rheumatoid arthritis, and osteoarthritis.

The aim of the present study was to determine synovial levels of ELR (+) CXC chemokines, known to attract mainly neutrophils to inflamed tissues by binding the neutrophil chemokine receptors CXCR1 and CXCR2 and promoting neovascularization in patients with various inflammatory disorders. The study group consisted of 14 patients with Behçet's disease and nine with familial Mediterranean fever. Fourteen patients with rheumatoid arthritis and 16 with osteoarthritis served as controls. Synovial chemokine levels were measured by two-step sandwich enzyme-linked immunosorbent assay, and significant differences were found in the various chemokines studied. In addition to its angiogenic properties, increased synovial levels of interleukin-8 by attraction of more neutrophils to synovial fluids might also be responsible for the acute synovitis in patients with Behçet's disease. However, the absence of chronic changes with the eventual development of pannus and erosions might result from relatively lower expression of interleukin-8 and the transient, short-lived nature of the arthritis observed in these patients.

Adult↗

Measurement of beta-thromboglobulin connective tissue activating peptide-III platelet antigen concentrations in pathologic synovial fluids.

Concentrations of the human platelet proteins beta-thromboglobulin (beta-TG) and connective tissue activating peptide (CTAP)-III were measured in synovial fluid (SF) samples by radioimmunoassay of their common beta-T/CTAP antigen. beta-T/CTAP antigen concentrations were higher in rheumatoid SF than in samples from osteoarthritis patients. When concurrent SF and plasma samples were compared, SF antigen levels were weakly correlated with plasma antigen concentrations. Incubation of fresh polymorphonuclear leukocytes with antigen in SF resulted in apparent destruction of the antigen. Our study confirms the presence of a platelet-derived growth factor antigen in the pathologic joint, and suggests that leukocytes may participate in its clearance from inflamed joints.

Antigens↗

Immunochemical and ultrastructural characterization of serum proteins associated with monosodium urate crystals (MSU) in synovial fluid cells from patients with gout.

Immunoelectron microscopic (IEM) analysis of the surface coats of intracellular and extracellular monosodium urate (MSU) crystals in synovial fluid (SF) in gouty arthritis was performed using the ferritin-bridge method. Cells from patients with acute gout were fixed in 1% glutaraldehyde containing 0.05% saponin to permeabilize membranes for access of immunochemicals to intracellular antigens. Intracellular MSU crystals were observed in phagosomes of greater than 75% of both polymorphonuclear (PMNs) and mononuclear cells. Coating of crystals with IgG was more prominent than with IgM or IgA. Other proteins such as C3, and fibrinogen were also found to a lesser extent. Albumin was not detected in appreciable amounts on MSU crystals. Extracellular crystals also showed IgG to be bound more prominently than other proteins. The various proteins, shown here for the first time to be clearly associated with intracellular crystals by EM, and other materials associated with MSU crystals may influence the phlogistic properties of these crystals.

Arthritis↗

Pathogen antigen- and superantigen-reactive synovial fluid T cells in reactive arthritis.

Analysis of pathogen-reactive T cell clones (CD3+4+8-TCR alpha beta +), isolated from the synovial fluid of 2 HLA-B27-positive patients with Yersinia enterocolitica-triggered reactive arthritis, has provided important information about the cellular immune response to this disease-inciting pathogen. This study demonstrates that the proteins secreted by Y. enterocolitica, including a protein with tyrosine phosphatase activity (YopH), are potent immunogens stimulating CD4+ cells within the inflamed joint. The pathogen-reactive T cell clones preferentially utilized a limited set of T cell receptor variable region gene segments. A purified Yersinia superantigen triggered a proliferative response in most of the antigen-reactive T cell clones tested. These results suggest that the activity of this pathogen's superantigen influences the cellular immune response to its antigens.

Adult↗

Local anti-type II collagen antibody production in rheumatoid arthritis synovial fluid. Evidence for an HLA-DR4-restricted IgG response.

OBJECTIVE: To investigate the production of type II collagen (CII) antibodies in the synovial fluid (SF) of rheumatoid arthritis (RA) patients, and to examine the HLA dependence of this local production. METHODS: The ELISPOT method was used for enumerating anti-CII-reactive cells. Serologic tissue typing was performed. RESULTS: Anti-CII-reactive cells were found in the SF of 16 of 31 patients, but not in any of the peripheral blood samples obtained in parallel. SF anti-CII antibody production showed no correlation with clinical parameters, but its frequency increased significantly with age. The IgG anti-CII response occurred exclusively in patients who were positive for HLA-DR4 and was significantly associated with DR4. CONCLUSION: Anti-CII production may be important in local immune complex formation. The indirect demonstration of a DR4-restricted T cell response to CII is an indication of a pathogenetic role of collagen autoimmunity in RA.

Adult↗

Phospholipase A2 activity in sera and synovial fluids in rheumatoid arthritis and osteoarthritis. Its possible role as a proinflammatory enzyme.

Phospholipase A2 (PLA2) activity was found in the sera and synovial fluids (SF) in rheumatoid arthritis (RA) and osteoarthritis (OA). PLA2 activity in RA SF was 6158 +/- 549 (SEM) U/ml (n = 48) and in RA sera 554 +/- 175 U/ml (normal sera-115 +/- 12 U/ml). In OA SF PLA2 activity was 5069 +/- 542 U/ml (n = 28), and in OA sera 268 +/- 55 U/ml. There was no significant difference between SF PLA2 activity in RA and OA. PLA2 activity in SF did not correlate with muramidase (lysozyme), beta-glucuronidase, total protein or white cell count, which were all significantly higher in RA SF than OA. A positive correlation between PLA2 in SF and matched sera was found in both RA and OA. It may be concluded that significant elevation of extracellular PLA2 occurs in both RA and OA, especially in the SF. The fact that high PLA2 did not correlate with other enzymes such as lysozyme and beta-glucuronidase, which are usually high in RA and low in OA SF, may mean that the handling of PLA2 in the joint space is different from other enzymes.

Arthritis, Rheumatoid↗