chondroitin-sulfates and Arthritis--Rheumatoid

The rheumatic diseases have been associated with accelerated atherosclerosis. Rheumatoid arthritis (RA) is a systemic inflammatory disease characterized by persistent synovial inflammation which leads to disability and structural changes in joints. Epidemiological studies have demonstrated an increased cardiovascular mortality in patients with RA. In these patients, atherosclerotic plaque occurs earlier, and it has a faster evolution than in general population. Atherosclerosis (AT) is also an inflammatory disease partly mediated by cytokines, many of them involved on chronic synovitis. Our group has developed a rabbit experimental model of AT aggravated by chronic arthritis to study inflammatory mechanisms involved on the progression of vascular lesions and their response to drugs. A preliminary study using this model suggests a beneficial effect of chondroitin sulphate (CS), a drug recommended for the treatment of osteoarthritis, in controlling AT lesions. Yet clinical trials should be conducted with this compound to address the same hypothesis in human studies.

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Atherosclerosis; Chondroitin Sulfates; Humans; Osteoarthritis; Rabbits

Topics: Arthritis, Rheumatoid; Cartilage, Articular; Cathepsins; Chondroitin Sulfates; Collagen; Cyclooxygenase Inhibitors; Glycoproteins; Glycosaminoglycans; Humans; Osteoarthritis; Peptide Hydrolases; Prostaglandins; Protease Inhibitors; Proteoglycans; Water

Inter alpha inhibitor (IαI) is an abundant serum protein consisting of three polypeptides: two heavy chains (HC1 and HC2) and bikunin, a broad-specificity Kunitz-type proteinase inhibitor. The complex is covalently held together by chondroitin sulfate but during inflammation IαI may interact with TNF-stimulated gene 6 protein (TSG-6), which supports transesterification of heavy chains to hyaluronan. Recently, IαI was shown to inhibit mouse complement in vivo and to protect from complement-mediated lung injury but the mechanism of such activity was not elucidated. Using human serum depleted from IαI, we found that IαI is not an essential human complement inhibitor as was reported for mice and that such serum has unaltered hemolytic activity. However, purified human IαI inhibited classical, lectin and alternative complement pathways in vitro when added in excess to human serum. The inhibitory activity was dependent on heavy chains but not bikunin and detected at the level of initiating molecules (MBL, properdin) in the lectin/alternative pathways or C4b in the classical pathway. Furthermore, IαI affected formation and assembly of the C1 complex and prevented assembly of the classical pathway C3-convertase. Presence and putative interactions with TSG-6 did not affect the ability of IαI to inhibit complement thus implicating IαI as a potentially important complement inhibitor once enriched onto hyaluronan moieties in the course of local inflammatory processes. In support of this, we found a correlation between IαI/HC-containing proteins and hemolytic activity of synovial fluid from patients suffering from rheumatoid arthritis.

Topics: Alpha-Globulins; Animals; Arthritis, Rheumatoid; Cell Adhesion Molecules; Chondroitin Sulfates; Complement Activation; Complement System Proteins; Female; Humans; Hyaluronic Acid; Lung Injury; Male; Mice; Synovial Fluid

To study efficacy of the chondroprotector chondroitin sulphate (structum, Pier Fabr Medicament Production, France) in patients with rheumatoid arthritis (RA) and secondary osteoarthrosis of the knee joints.. 15 women with a long history of RA (mean duration 11.9 years) entered an open non-randomized trial of structum. The patients had a severe progressive highly active RA with a definite x-ray stage of the disease. 13 patients had a positive rheumatoid factor (1:80 to 1:1280) and involved knee joints which had been affected for 1 to 10 years (mean 5.3 years). The second x-ray stage was in 8 patients, the third stage of knee joints arthrosis was in 7 ones. A marked pain syndrome in the knee joints upon movement (mean 64.7 mm by VAS) was observed in all the examinees and at rest (mean 28 mm by VAS) in 13 of 15 patients. Structum was given according to a standard scheme: 500 mg 3 times a day for 3 weeks than 500 mg 2 times a day for up to 6 months. Basic drugs for RA were the same for all the observation period.. Structum noticeably improved knee joint function (mean Leken's index 12.8, 11.3 and 9.4 scores before the treatment, on treatment month 3 and 6. Movement pain syndrome VAS reduced from 64.7 mm at the start to 51 mm 3 months and 37.5 mm 6 months later, rest VAS--from 19 to 10.3 and 6.4 mm, respectively. The demand in intraarticular glucocorticoids went down from 52 injections at the start of therapy to 6 after 6 months. Side effects for 6 months were absent. Overall efficacy was good (73.3% and 80%) as judged by the doctors and patients, respectively. After 6 months of therapy control x-rays found no progression of destructive changes in the knee joints (by MRI--in 4 patients).. Structum has a marked positive therapeutic effect in patients with severe and long-term course of RA with associated pronounced secondary joint arthrosis.

Topics: Adult; Arthritis, Rheumatoid; Chondroitin Sulfates; Drug Therapy, Combination; Female; Glucocorticoids; Humans; Injections, Intra-Articular; Knee Joint; Middle Aged; Osteoarthritis, Knee; Radiography; Rheumatoid Factor

Long-term oral tofacitinib (TOF) administration has been linked to serious side effects majorly immunological suppression. The aim of this work was to enhance the therapeutic efficacy of TOF by chondroitin sulphate (CS) coated proglycosomes through the anchoring of high-affinity CS to CD44 receptors on immune cells in the inflammatory region. The CS was coated onto the TOF-loaded proglycosomes (CS-TOF-PG) formulations and they were evaluated for in vitro drug release, ex vivo (permeation, dermatokinetics) studies. In vivo efficacy studies were carried out in Freund's complete adjuvant (CFA) induced arthritis model. The optimized CS-TOF-PG showed particle sizes of 181.13 ± 7.21 nm with an entrapment efficiency of 78.85 ± 3.65 %. Ex-vivo studies of CS-TOF-PG gel exhibited 1.5-fold high flux and 1.4-fold dermal retention compared to FD-gel. The efficacy study revealed that CS-TOF-PG showed a significant (P < 0.001) reduction in inflammation in arthritic rat paws compared to the TOF oral and FD gel. The current study ensured that the CS-TOF-PG topical gel system would provide a safe and effective formulation for localization and site-specific delivery of TOF at the RA site and overcome the adverse effects associated with the TOF.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Chondroitin Sulfates; Piperidines; Rats

Rheumatoid arthritis is a chronic autoimmune disease characterised by joint synovial inflammation, along with cartilage and bone tissue destruction. Dendrimers can offer new opportunities as drug delivery systems of molecules of interest. Herein we aimed to develop poly(amidoamine) dendrimers (PAMAM), functionalised with chondroitin sulphate (CS), lined with anti-TNF α antibodies (Abs) to provide anti-inflammatory properties. Physicochemical characterisation demonstrated that anti-TNFα Abs-CS/PAMAM dendrimer NPs were successfully produced. The in vitro studies revealed that CS/PAMAM dendrimer NPs did not affect the ATDC5 and THP-1 cell lines' metabolic activity and proliferation, presenting good cytocompatibility and hemocompatibility. Moreover, anti-TNFα Abs-CS/PAMAM dendrimer NPs showed suitable TNF α capture capacity, making them appealing for new immunotherapies in RA patients.

Topics: Arthritis, Rheumatoid; Chondroitin Sulfates; Dendrimers; Humans; Tumor Necrosis Factor-alpha

This research aims to coat Teriflunomide (TEF) loaded conventional nanoliposomes (CON-TEF-LIPO) with Chondroitin sulphate (CS) to produce CS-TEF-LIPO for the effective treatment of Rheumatoid arthritis (RA). Both CON-TEF-LIPO and CS-TEF-LIPO were produced, characterized and evaluated for their active targeting potential towards CD44 receptors. Cell cytotoxicity, cell viability and intracellular uptake study on differentiated U937 and MG-63 cells demonstrated the active targeting of CS-TEF-LIPO towards CD44 receptors. Furthermore, in vivo pharmacodynamic, biochemical, radiological and histopathological studies performed in adjuvant induced arthritic (AIA) rat model showed a significant (P < 0.05) reduction in inflammation in arthritic rat paw in CS-TEF-LIPO group compared to TEF and CON-TEF-LIPO groups. Moreover, liver toxicity study revealed that CS-TEF-LIPO showed no signs of toxicity and biodistribution study revealed the accumulation of CS-TEF-LIPO in synovial region of arthritic rat. Taken together, results suggest that CS-TEF-LIPO could provide a new insight for an effective treatment of RA.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Chondroitin Sulfates; Crotonates; Glioma; Humans; Hydroxybutyrates; Liposomes; Male; Nanoparticles; Nitriles; Rats; Rats, Wistar; Tissue Distribution; Toluidines; Tumor Cells, Cultured

To investigate selenium (Se) concentrations in serum of patients with rheumatoid arthritis (RA), osteoarthritis (OA), and Kashin-Beck disease (KBD), together with the effect of Se supplement (chondroitin sulfate [CS] nano-Se [SeCS]) on CS structure-modifying sulfotransferases in KBD chondrocyte. Fifty serum samples from each group with aged-matched (40-60 years), normal control (N), RA, OA, and KBD (25 males and females, respectively) were collected to determine Se concentrations. Furthermore, the KBD chondrocytes were divided into two groups following the intervention for 24 h: (a) non-treated KBD group and (b) SeCS-treated KBD group (100 ng/mL SeCS). The ultrastructural changes in chondrocytes were observed by transmission electron microscopy (TEM). Live/dead staining was used to observe cell viability. The expression of CS-modifying sulfotransferases including carbohydrate sulfotransferase 12, 13, and 15 (CHST-12, CHST-13, and CHST-15, respectively), and uronyl 2-O-sulfotransferase (UST) were examined by quantitative real-time polymerase chain reaction and western blotting analysis after SeCS intervention. The Se concentrations in serum of KBD, OA, and RA patients were lower than those in control. In OA, RA, and control, Se concentrations were higher in male than in female, while it is opposite in KBD. In the cell experiment, cell survival rate and mitochondrial density were increased in SeCS-treated KBD groups. Expressions of CHST-15, or CHST-12, and CHST-15 on the mRNA or protein level were significantly increased. Expression of UST slightly increased on the mRNA level, but no change was visible on the protein level. Se deficiency in serum of RA, OA, and KBD was observed. SeCS supplemented in KBD chondrocytes improved their survival rate, ameliorated their ultrastructure, and increased the expression of CS structure-modifying sulfotransferases.

Topics: Adult; Arthritis, Rheumatoid; Chondrocytes; Chondroitin Sulfates; Female; Humans; Kashin-Beck Disease; Male; Middle Aged; Osteoarthritis; Selenium

Oral treatment of rheumatoid arthritis (RA) with the immunomodulator, leflunomide (LEF), is associated with systemic side effects namely immunosuppression and hepatotoxicity. Herein, attempts to improve LEF therapeutic outcomes via nanostructured lipid carriers (NLCs) targeting inflamed rheumatic joints were executed. LEF-NLCs coated with either chondroitin sulphate (CHS) or chitosan (CS) were around 250 nm in size with negative or positive charge, respectively. Particle coating was evidenced by TEM and FTIR analysis. NLCs generally ensured sustained release profile up to 21 days, particularly extended in coated formulations. In vivo pharmacokinetic study of LEF suspension, uncoated NLCs, CHS- and CS-coated NLCs was carried out. Following oral administration in RA-induced rat model, joint diameter, paw inflammation, liver functions were measured, in addition to histological examination of liver, kidney and joints. Results revealed improved joint healing and reduced hepatotoxicity following treatment with nanoencapsulated LEF compared to LEF suspension, whereby CHS-NLCs ensued the highest C

Topics: Administration, Oral; Animals; Arthritis, Rheumatoid; Chitosan; Chondroitin Sulfates; Drug Carriers; Drug Liberation; Immunosuppressive Agents; Joints; Leflunomide; Lipids; Male; Nanostructures; Rats, Sprague-Dawley

The study aimed to investigate the effect of the oral administration for 15 days of either Echinacea (E) or genuphil (a composite of chondroitin sulphate, glucosamine and methyl sulfonyl methane [GCM]) nutraceutical supplements on female rat model of acute or chronic arthritis induced by bacterial outer membrane protein (OMP) from faecal flora of healthy and rheumatic humans. Anti-cyclic citrullinated peptide (anti-CCP2), C-reactive protein (CRP) and rheumatoid factor (RF) values increased (p < 0.05) in both arthritic groups as compared to normal values. The rheumatic markers anti-CCP2, CRP and RF values decreased significantly in E- and GCM-treated groups compared to arthritic none-treated acute or chronic groups. The results of RF values of GCM-treated groups in acute and chronic models decreased exhibiting no statistical difference compared with the normal value. Histological examinations of the hind paw sections revealed moderate inflammation, oedema and mild proliferation of synovial cells in acute arthritic rats and more damage to cartilage and bone with severe inflammation in chronic ones. Echinacea acute treated group showed edema with proliferated synovial membrane and partial damage in cartilage and bone. While in the E-chronic treated group, rough edge with destructed cartilage and bone existed. However, the acute GCM group revealed mild cartilage damage. But the chronic GCM group showed mild synovial cells proliferation and revealed no inflammation with mild cartilage damage edge. Results demonstrated the OMP arthropathic property and through promising light on arthritis treatment using E- or GCM, with the advantage of GMC results over that of E-. The composite GCM is needed for further studies over the dose and duration to assess its preventive effects against the bacterial OMP arthrogenicity.

Topics: Animals; Antirheumatic Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Chondroitin Sulfates; Complex Mixtures; Dietary Supplements; Dimethyl Sulfoxide; Disease Models, Animal; Echinacea; Female; Glucosamine; Lysosomes; Plant Extracts; Rats; Stifle; Sulfones

Mesenchymal stem cells (MSCs) have the potential to differentiate into distinct mesenchymal tissues; including cartilage and bone, they can be an attractive cell source for cartilage tissue engineering approaches. Our objective here was to compare the in vitro chondrogenic potential of MSCs isolated from patients with rheumatoid arthritis (RA) and osteoarthritis (OA) with cells from normal donors.. Marrow samples were removed during bone surgery and adherent cell cultures were established. The cells were then passed into a newly developed microaggregate culture system in a medium containing transforming growth factor beta3, insulin, dexamethasone and/or demineralized bone matrix. In vitro chondrogenic activity was measured as metabolic sulfate incorporation and type II collagen expression in pellet cultures.. Culture-expanded MSCs from RA and OA patients did not differ significantly from the normal population with respect to their chondrogenic potential in vitro. Capability of total protein and proteoglycan synthesis as well as collagen II mRNA expression by cell aggregates was similar for all cell preparations in the presence of the appropriate growth and differentiation factors. Chondroprotective drugs such as chondroitin sulfate and glucosamine enhanced, whereas chloroquine inhibited chondrogenesis in normal donor-derived or patient-derived MSC cultures. Galectin-1, a beta-galactoside-binding protein with marked anti-inflammatory activity, stimulated the chondrogenic differentiation of mesenchymal cells in low (<2 microg/ml) concentration.. These findings show that MSCs from RA and OA patients possess similar chondrogenic potential as MSCs isolated from healthy donors, therefore these cells may serve as a potential new prospect in cartilage replacement therapy.

Topics: Adipogenesis; Aged; Arthritis, Rheumatoid; Bone Marrow Cells; Cell Differentiation; Cells, Cultured; Chondrogenesis; Chondroitin Sulfates; Collagen Type II; Female; Galectins; Glucosamine; Humans; Male; Mesenchymal Stem Cells; Microscopy, Electron, Transmission; Middle Aged; Osteoarthritis; Osteogenesis; Reverse Transcriptase Polymerase Chain Reaction

Topics: Acetates; Antirheumatic Agents; Arthritis, Rheumatoid; Chondroitin Sulfates; Clinical Trials as Topic; Drug Therapy, Combination; Humans; Osteoarthritis; Piperidines; Pyrimidines; Pyrroles

Serum hyaluronan (HA) and chondroitin sulfate (CS) epitopes WF6 and 3B3 (+) were determined to investigate disease association in patients with osteoarthritis (OA), rheumatoid arthritis (RA) and healthy controls.. Specific assays for HA and CS epitopes WF6 and 3B3 (+) were established and applied to a cross-sectional study of serum samples from patients (96 OA, 57 RA and 50 healthy controls).. Both CS epitopes were increased in serum of many OA and RA patients and average levels were significantly above in healthy controls. In contrast serum HA was increased in RA, but only in few OA patients.. CS epitopes WF6 and 3B3 (+) are raised in serum of patients with both OA and RA and were thus distinct from serum HA. The results suggest that OA may be detected systemically as well as RA. The range of levels of CS epitopes detected in OA and RA was wide and correlation with any aspect of disease activity is yet to be determined.

Topics: Adult; Aged; Antibodies, Monoclonal; Arthritis, Rheumatoid; Biomarkers; Chondroitin Sulfates; Cross-Sectional Studies; Epitopes; Female; Humans; Hyaluronic Acid; Male; Middle Aged; Osteoarthritis

Intraperitoneally administered chondroitin sulfate E inhibited the development of antibody-induced arthritis, a model of rheumatoid arthritis, while chondroitin 4-sulfate showed no effects. Chondroitin sulfate E inhibited in vitro differentiation of osteoclasts, which play key roles in the etiology of rheumatoid arthritis. One of the targets of chondroitin sulfate E is midkine, a heparin-binding growth factor or cytokine. Indeed, a chimeric-type siRNA for midkine inhibited the development of antibody-induced arthritis and adhesion of the omentum to the injured abdominal wall. These results indicate the significance of midkine as a molecular target to treat or prevent rheumatoid arthritis and adhesion after surgery, and the utility of chondroitin sulfate E to inhibit midkine in vivo.

Topics: Animals; Antibodies; Arthritis, Rheumatoid; Cell Differentiation; Chondroitin Sulfates; Cytokines; Disease Models, Animal; Female; Injections, Intraperitoneal; Mice; Mice, Inbred BALB C; Midkine; Osteoblasts; RNA, Small Interfering

Thrombin is a key factor in the stimulation of fibrin deposition, angiogenesis, proinflammatory processes, and proliferation of fibroblast-like cells. Abnormalities in these processes are primary features of rheumatoid arthritis (RA) in synovial tissues. Tissue destruction in joints causes the accumulation of large quantities of free hyaluronic acid (HA) in RA synovial fluid. The present study was conducted to investigate the effects of HA and several other glycosaminoglycans on antithrombin, a plasma inhibitor of thrombin. Various glycosaminoglycans, including HA, chondroitin sulfate, keratan sulfate, heparin, and heparan, were incubated with human antithrombin III in vitro. The residual activity of antithrombin was determined using a thrombin-specific chromogenic assay. HA concentrations ranging from 250 to 1000 mug/ml significantly blocked the ability of antithrombin to inhibit thrombin in the presence of Ca2+ or Fe3+, and chondroitin A, B and C also reduced this ability under the same conditions but to a lesser extent. Our study suggests that the high concentration of free HA in RA synovium may block antithrombin locally, thereby deregulating thrombin activity to drive the pathogenic process of RA under physiological conditions. The study also helps to explain why RA occurs and develops in joint tissue, because the inflamed RA synovium is uniquely rich in free HA along with extracellular matrix degeneration. Our findings are consistent with those of others regarding increased coagulation activity in RA synovium.

Topics: Antithrombin III; Arthritis, Rheumatoid; Autoimmune Diseases; Calcium; Chondroitin Sulfates; Chromogenic Compounds; Dermatan Sulfate; Glycosaminoglycans; Heparin; Heparitin Sulfate; Humans; Hyaluronic Acid; Iron; Keratan Sulfate; Synovial Fluid; Thrombin

Rheumatoid arthritis (RA) is a chronic, systemic, and inflammatory disease of connective tissue with unknown etiology. We investigated whether aberrant immune responses to glycosaminoglycans (GAGs), a major component of joint cartilage, joint fluid, and other soft connective tissue, causes this disease. Here we show that injection of GAGs such as hyaluronic acid, heparin, and chondroitin sulfates A, B, and C induce arthritis, tendosynovitis, dermatitis, and other pathological conditions in mice. We developed a technique by staining tissue specimens with fluorochrome- or biotin-labeled GAGs to visualize the direct binding between cells and GAGs. We discovered that inflammatory infiltrates from the affected tissue are dominated by a distinct phenotype of GAG-binding cells, a significant portion of which are CD4(+) T cells. GAG-binding cells seem to be expanded in bone marrow of GAG-immunized mice. Furthermore, we identified GAG-binding cells in inflamed synovial tissue of human patients with RA. Our findings suggest that carbohydrate self-antigenic GAGs provoke autoimmune dysfunctions that involve the expansion of GAG-binding cells which migrate to anatomical sites rich in GAGs. These GAG-binding cells might, in turn, promote the inflammation and pathology seen both in our murine model and in human RA.

Topics: Animals; Arthritis, Rheumatoid; Autoantibodies; Bone Marrow Cells; CD4-Positive T-Lymphocytes; Chondroitin Sulfates; Dermatan Sulfate; Disease Models, Animal; Female; Glycosaminoglycans; Heparin; Humans; Hyaluronic Acid; Immunophenotyping; Mice; Mice, Inbred C57BL

The therapeutic effect of a nutritional supplement consisting of a combination of glucosamine hydrochloride (FCHG49), purified sodium chondroitin sulfate (TRH122), and manganese ascorbate (GCM)3 was investigated in the rat model of collagen-induced autoimmune arthritis (CIA). The GCM compound was mixed with a palatable nutritional paste (Nutri-cal [NC]). Oral administration of the NC/GCM compound was initiated in 26 rats 10 days before immunization and continued until the day of sacrifice. One group of 12 control rats was given no oral agents; a second group of 12 control rats received NC only. Evaluations included arthritis index (AI) scoring by three independent evaluators, histologic index (HI) scoring of lesions, T-cell proliferation, and serological studies for antibody classes and subclasses. Both the AI and HI criteria showed a statistically significant reduction in the prevalence of CIA in rats pretreated with the NC/GCM (54%) compared to the combined control groups (96%, chi2 analysis P = 0.001). Rats fed the NC/GCM also exhibited a significant decrease in the severity of autoimmune arthritis in both the AI and HI compared to control Group 2 (immunized-NC) (chi2 analysis P < 0.05). Histological studies verified the decreased incidence of arthritis in the NC/GCM group compared to control Group 2. GCM treatment failed to alter T-cell proliferation and antibody production to bovine type-II collagen, indicating that its effects are not due to alteration of the antigen-specific immune response.

Topics: Animals; Arthritis, Rheumatoid; Ascorbic Acid; Autoantibodies; Chondroitin Sulfates; Collagen; Deferoxamine; Dietary Supplements; Drug Therapy, Combination; Female; Glucosamine; Knee; Manganese; Organometallic Compounds; Rats; Severity of Illness Index

Cathepsin K is the predominant cysteine protease in osteoclast-mediated bone remodeling, and the protease is thought to be involved in the pathogenesis of diseases with excessive bone and cartilage resorption. Osteoclastic matrix degradation occurs in the extracellular resorption lacuna and upon phagocytosis within the cell's lysosomal-endosomal compartment. Since glycosaminoglycans (GAGs) are abundant in extracellular matrixes of cartilage and growing bone, we have analyzed the effect of GAGs on the activity of bone and cartilage-resident cathepsins K and L and MMP-1. GAGs, in particular chondroitin sulfates, specifically and selectively increased the stability of cathepsin K but had no effect on cathepsin L and MMP-1. GAGs strongly enhanced the stability and, to a lesser extent, the catalytic activity of cathepsin K. To combine the activity and stability parameters, we defined a novel kinetic term, named cumulative activity (CA), which reflects the total substrate turnover during the life span of the enzyme. In the presence of chondroitin-4-sulfate (C-4S), the CA value increased 200-fold for cathepsin K but only 25-fold with chondroitin-6-sulfate (C-6S). C-4S dramatically increased the hydrolysis of soluble as well insoluble type I and II collagens, whereas the effects of C-6S and hyaluronic acid were less pronounced. C-4S acts in a concentration-dependent manner but reaches saturation at approximately 0.1%, a concentration similar to that found in the synovial fluid of arthritis patients. C-4S increased the cathepsin K-mediated release of hydroxyproline from insoluble type I collagen 10-fold but had only a less than 2-fold enhancing effect on the hydrolysis of intact cartilage. The relatively small increase in the hydrolysis of cartilage by C-4S was attributed to the endogenous chondroitin sulfate content present in the cartilage. Although C-4S increased the pH stability at neutral pH, a significant increase in the collagenolytic activity of cathepsin K at this pH was not observed, thus suggesting that the unique collagenolytic activity of cathepsin K at acidic pH is mechanistically determined and not by the enzyme's instability at neutral pH. The selective and significant stabilization and activation of cathepsin K activity by C-4S may provide a rationale for a novel mechanism to regulate the enzyme's activity during bone growth and aging, two processes known for significant changes in the GAG content.

Topics: Arthritis, Rheumatoid; Cartilage, Articular; Cathepsin K; Cathepsin L; Cathepsins; Chondroitin Sulfates; Collagen; Connective Tissue Cells; Cysteine Endopeptidases; Endopeptidases; Extracellular Matrix; Glycosaminoglycans; Humans; Kinetics; Matrix Metalloproteinase 1; Recombinant Proteins

To investigate the correlation between synovial fluid (SF) concentrations of metalloproteinase (MMP) -2, -3, and -9, tissue inhibitors of metalloproteinases (TIMP)-1 and -2, chondroitin 4-, 6-sulfate (deltadi-C4S, deltadi-C6S), hyaluronic acid (deltadi-HA), antigenic keratan sulfate (KS), and radiologic grade in patients with rheumatoid arthritis (RA) and to assess the clinical value of these factors as disease markers.. Enzyme linked immunoassays and high performance liquid chromatography were used. SF samples were collected from 52 patients with RA. Radiographic (Larsen grade) and clinical evaluations were also done.. There was no significant correlation between the concentrations of MMP, TIMP, and deltadi-C4S, deltadi-C6S, deltadi-HA, and antigenic KS. No significant correlations were found between the radiologic grade and the concentrations of MMP or TIMP in SF. There was a significant increase in the concentration of antigenic KS and deltadi-C6S/deltadi-C4S ratio in SF from patients with Larsen grade 2 compared to grade 5. A significant correlation between deltadi-C6S and antigenic KS concentrations in SF was observed.. Although there were large variances between the samples in this study, proteoglycan metabolism in the cartilage matrix of the patients with RA might have been increased in the early phases of tissue destruction and decreased in advanced stages because of scanty cartilage tissue. The concentrations of MMP-2, -3, -9 and TIMP-1, -2 in SF did not seem to be influenced by the amount of residual cartilage.

Topics: Aged; Arthritis, Rheumatoid; Chondroitin Sulfates; Female; Glycosaminoglycans; Humans; Hyaluronic Acid; Keratan Sulfate; Male; Metalloendopeptidases; Middle Aged; Radiography; Synovial Fluid; Tissue Inhibitor of Metalloproteinases

The aims of the present rheumatoid arthritis (RA) study were (1) to examine the levels of serum 3-B-3 and 7-D-4 to find out whether they are different from controls, (2) to find out whether the concentrations of these epitopes change with disease duration in early RA and (3) whether the serum concentrations of 3-B-3 and 7-D-4 in early RA are prognostic for subsequent disease progression.. The concentrations of 3-B-3 and 7-D-4 in sera were quantitated by immunoassays.. The levels of 3-B-3 and 7-D-4 were significantly lower in RA than in controls (3- to 30-fold, P < 0.001). Changes in 3-B-3 and 7-D-4 were apparent with disease duration. At first presentation, the 3-B-3 concentration was lowest and increased at 12 months (3-fold, P < 0.001). This increase was transient since by 24 and 36 months the concentrations were not different to those at first presentation. The level of 7-D-4 was also lowest when the patients first presented at clinic and increased with time at 6 months (2-fold, P < 0.001). The increase was more prolonged for 7-D-4, remaining elevated at 12, 24 and 36 months. The lack of correlations of serum 3-B-3 and 7-D-4 with clinical measurements showed that these markers were not prognostic for disease severity.. The levels of 3-B-3 and 7-D-4 differed between RA and control sera, and changed with disease duration. These markers were not prognostic in predicting disease outcome.

Topics: Adult; Aged; Antibodies, Monoclonal; Arthritis, Rheumatoid; Chondroitin Sulfates; Disease Progression; Epitopes, B-Lymphocyte; Female; Humans; Immunoassay; Male; Middle Aged; Prognosis

This study investigated the correlation between temporomandibular joint (TMJ) disease and the composition of glycosaminoglycans (GAGs) components in the synovial fluid (SF).. Synovial fluid (SF) was obtained from 30 TMJs of 28 female patients diagnosed as having a displaced disc with reduction (WR) (seven joints), a displaced disc without reduction (WOR) (13 joints), osteoarthritis (OA) (five joints), or rheumatoid arthritis (RA) (five joints) by MR imaging and clinical examination. After the SF was directly aspirated, It was digested with chondroitinase ABC and hyaluronidase, and the concentration of unsaturated disaccharide isomers of chondroitin 6-sulfate (delta di-6S), chondroitin 4-sulfate (delta di-4S) and hyaluronic acid (delta di-HA) were measured by high-performance liquid chromatography (HPLC) combined with fluorometry. The ratio of delta di-6S or delta di-4S to delta di-HA, and delta di-6S to delta di-4S, were calculated.. There were no significant differences in concentrations of delta di-6S, delta di-4S, or delta di-HA among the groups. The ratio of delta di-6S to delta di-4S was 2.7 +/- 1.4 in OA, 2.6 +/- 0.9 in joints with WOR, 2.9 +/- 1.2 in joints with WR, and 1.3 +/- 0.4 in RA synovial fluid. Differences in the delta di-6S: delta di-4S ratio between RA and the other conditions were statistically significant (P < .05).. These results suggest that the delta di-6S:delta di-4S ratio in the synovial fluid of the TMJ reflects the proteoglycan metabolism of the joint tissues, particularly of the articular cartilage and synovial tissue. This ratio could be used to diagnose joint diseases and to predict articular cartilage destruction or synovial proliferation caused by these diseases.

Topics: Adult; Arthritis, Rheumatoid; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Disaccharides; Female; Fluorometry; Glycosaminoglycans; Humans; Hyaluronic Acid; Joint Dislocations; Middle Aged; Osteoarthritis; Paracentesis; Statistics, Nonparametric; Synovial Fluid; Temporomandibular Joint; Temporomandibular Joint Disc; Temporomandibular Joint Disorders

To develop a biodegradable, inflammation-responsive microsphere system for the intraarticular delivery of therapeutic proteins.. Microspheres were synthesized by complex coacervation. Radiolabeled protein release and microsphere degradation were assessed by exposing the microspheres to human synovial fluids (SF) and recombinant gelatinase. Microsphere degradation was confirmed by scanning electron microscopy (SEM). Microsphere biocompatibility was evaluated in vitro by incubating the microspheres with human synoviocytes, and in vivo by injection into mouse joints.. Optimal microsphere formulation was developed. Significant (up to 100%) release of encapsulated protein occurred in SF samples with measurable metalloprotease activity, while release was minimal in SF with negligible activity. Dissolution of microspheres exposed to gelatinase was confirmed by SEM. Microspheres were found to be noncytotoxic in vitro, and noninflammatory in vivo.. Microsphere encapsulation is an inflammation-responsive and biocompatible system of protein delivery that holds promise for use in the delivery of therapeutic proteins to the joint.

Topics: Arthritis, Rheumatoid; Biocompatible Materials; Chondroitin Sulfates; Collagenases; Drug Delivery Systems; Gelatin; Gelatinases; Humans; Hydrogen-Ion Concentration; Injections, Intra-Articular; Kinetics; Metalloendopeptidases; Microspheres; Osteoarthritis; Synovial Fluid

To determine concentrations of chondroitin sulphate (CS) and keratan sulphate (KS) epitopes, glycosaminoglycans (GAGs) and hyaluronan (HA) in knee synovial fluid (SF) from normal subjects and patients with osteoarthritis (OA) or rheumatoid arthritis (RA), to test whether these variables may be used as markers of the OA process.. OA was subdivided into large joint OA (LJOA), nodal generalised OA (NGOA), and OA with calcium pyrophosphate crystal deposition (CPA). Clinical assessment of inflammation (0-6) was undertaken on OA and RA knees. Knee SF was examined by enzyme linked immunosorbent assay for: CS epitopes, using monoclonal antibodies 3-B-3 and 7-D-4; KS epitope using monoclonal antibody 5-D-4; and HA, using biotinylated HA binding region of cartilage proteoglycan. Total sulphated GAGs were measured by dye binding with 1:9 dimethylmethylene blue.. Increased SF 3-B-3 concentrations and 3-B-3/GAG ratio were found in OA, compared with RA or normal knees, with higher 3-B-3 and 3-B-3/GAG in LJOA and NGOA than in CPA. SF 7-D-4 and 7-D-4/GAG were reduced in RA, compared with normal and OA; SF 5-D-4 was reduced in OA compared with normal. GAG and HA concentrations were decreased in both OA and RA. No correlations with radiographic scores were observed, but SF 7-D-4 was lower in 'inflamed' compared with 'non-inflamed' RA and OA knees. In patients with bilateral samples there were strong correlations between right and left knees for all SF variables.. Changed concentrations of SF CS and KS can be detected in OA with a profile that differs from that seen in RA. Clinical subgrouping and local joint inflammation may influence these measures, supporting different pathogenesis within OA subgroups and requirement for careful patient characterisation in SF studies.

Topics: Adult; Aged; Arthritis, Rheumatoid; Biomarkers; Chondroitin Sulfates; Enzyme-Linked Immunosorbent Assay; Epitopes; Female; Glycosaminoglycans; Humans; Hyaluronic Acid; Keratan Sulfate; Knee Joint; Male; Middle Aged; Osteoarthritis; Synovial Fluid

Serum concentrations of specific cartilage and bone molecules reflecting tissue turnover were measured in two well-defined patient groups with early rheumatoid arthritis with distinctly different disease outcome to see if early differences in their levels are prognostic of the rate of joint destruction. Compared with a matched normal population, increased concentrations of cartilage oligomeric matrix protein (COMP) were found in all patients who developed rapid hip joint destruction. In contrast, levels of a putative marker of cartilage aggrecan synthesis, the chondroitin sulfate epitope 846, were increased only in patients with slow joint destruction. Levels of bone sialoprotein (BSP) were increased in both groups, as were levels of the C-propeptide of type II procollagen (CPII), a marker of collagen II synthesis. The increased concentrations of the 846 epitope in patients with slow joint destruction suggest increased aggrecan synthesis. The low levels of the 846 epitope in patients with rapid joint destruction, concomitant with elevated levels of CPII, suggest a selective increase in collagen synthesis. The elevated BSP levels indicate an increased bone turnover in both groups. Thus elevated serum levels of COMP may indicate an unfavorable prognosis for rapid joint destruction, whereas elevated 846 epitope indicates a more favorable prognosis.

Topics: Adult; Aged; Aggrecans; Arthritis, Rheumatoid; Bone and Bones; Calcium-Binding Proteins; Cartilage; Cartilage Oligomeric Matrix Protein; Chondroitin Sulfates; Collagen; Collagen Type II; Epitopes; Extracellular Matrix Proteins; Female; Glycoproteins; Humans; Hyaluronic Acid; Integrin-Binding Sialoprotein; Lectins, C-Type; Male; Matrilin Proteins; Middle Aged; Proteoglycans; Radiography; Radioimmunoassay; Sialoglycoproteins

The metabolism of the cartilage proteoglycan aggrecan was studied in patients with osteoarthritis (OA, n = 83), rheumatoid arthritis (RA, n = 127), and in controls (n = 117) using monoclonal antibody-based radioimmunoassays for glycosaminoglycans in the serum and synovial fluid (SF) to detect epitope 846 on chondroitin sulfate (probably only on recently synthesized molecules) and a keratan sulfate (KS) epitope AN9PI, present on intact and degraded molecules. Epitope 846 levels were always elevated in SF over serum (mean 38-fold in OA and 8.6-fold in RA) being highest in OA patients with the longest disease duration and greatest loss of cartilage, and lowest in RA joints with high leucocyte counts. Serum levels were more often elevated in RA (56%) than in OA (19%) and probably reflect increased aggrecan synthesis in diseased joints. KS levels were higher in SF than in serum in 69% of patients (up to 2.3-fold); levels were inversely (OA) and directly (RA) related to SF leucocyte counts. Serum KS was reduced in both diseases and in RA was inversely related to both systemic and joint inflammation markers. SF 846 levels were inversely related to SF KS in both diseases. These epitopes may provide a measure of the balance between cartilage synthesis and degradation in these diseases.

Topics: Adult; Aged; Aged, 80 and over; Aggrecans; Arthritis, Rheumatoid; Cartilage; Chondroitin Sulfates; Epitopes; Extracellular Matrix Proteins; Female; Humans; Keratan Sulfate; Lectins, C-Type; Male; Middle Aged; Osteoarthritis; Proteoglycans; Synovial Fluid

The localization and the nature of glycosaminoglycans (GAGs) in the synovial membrane in 30 patients with rheumatoid arthritis (RA) involving 40 knees were studied by newly developed histochemical methods. To detect the acidic glycoconjugates, sensitized diamine procedures were employed based upon high and low iron diamine stainings. To identify the various molecular species of the GAGs, enzyme (chondroitinase ABC and B, testicular hyaluronidase and keratanase) digestion and chemical modification (nitrous acid treatment) procedures were performed prior to the diamine stainings. The sensitized diamine methods could clearly stain the acidic glycoconjugates contained in the synovial tissue components in shades of brown to black, and could detect the precise distribution patterns of the GAGs. The results obtained in the present study confirmed that the tissue in RA synovial membranes contained various amounts of each GAG molecular species such as dermatan sulfate, chondroitin sulfate A/C, hyaluronic acid and heparan sulfate. Furthermore, the distribution patterns of dermatan and chondroitin sulfates in the diseased synovial tissues were pathophysiologically interesting; in the inflammatory areas, the molecular species of GAGs was primarily dermatan sulfate, whereas in the fibrotic areas, it was mainly chondroitin sulfate A/C. Such results appear to be useful for pathophysiological studies on the synovial tissues of RA.

Topics: Adult; Aged; Arthritis, Rheumatoid; Chondroitin Sulfates; Dermatan Sulfate; Female; Glycosaminoglycans; Heparitin Sulfate; Histocytochemistry; Histological Techniques; Humans; Hyaluronic Acid; Knee Joint; Male; Middle Aged; Synovial Membrane

Chondroitin sulphate is the major sulphated glycosaminoglycan present in the extracellular matrix of soft connective tissues and the aim of this study was to investigate the distribution of chondroitin sulphate species in normal and diseased synovium.. Distribution of chondroitin-4-sulphate/dermatan sulphate (Ch4S/DS) and chondroitin-6-sulphate in normal (n = 6), osteoarthritic (n = 4) and rheumatoid (n = 10) synovium was determined using an immunoperoxidase technique and specific monoclonal antibodies to chondroitinase ABC-digested preparations.. Ch4S/DS was expressed throughout the interstitium of all tissues and was also present on blood vessels in rheumatoid samples only. Ch6S was expressed in the lining layer of normal synovium but was absent from this site in osteoarthritic and rheumatoid tissues. Ch6S was also present on all blood vessels in all tissues.. The distinct zonal distributions of Ch4S/DS and Ch6S and their alteration in disease suggest these molecules have different and specific functions in normal and diseased synovium.

Topics: Arthritis, Rheumatoid; Chondroitin; Chondroitin Sulfates; Dermatan Sulfate; Humans; Immunoenzyme Techniques; Osteoarthritis; Synovial Membrane

The present study was undertaken to identify the cartilage matrix molecules that are bound with intermolecular disulfide bonds to IgG and serum albumin molecules recovered from the articular cartilage of patients with rheumatoid arthritis (RA) or osteoarthritis (OA). The cartilage specimens were extracted sequentially with three changes of neutral buffer, three changes of 6 M guanidine hydrochloride and then partially degraded with bacterial collagenase. The extracted IgG and albumin, along with matrix molecules bound to these proteins, were isolated with affinity chromatography using antibodies to IgG or human serum albumin conjugated to agarose beads. The isolated materials were characterized with sodium dodecyl sulfate polyacrylamide gel electrophoresis and transfer blotting, using specific antibodies to IgG, albumin, and proteoglycans. In the isolated materials, heteropolymers with IgG or albumin were identified. These polymers contained keratan sulfate and less frequently chondroitin-4-sulfate and chondroitin-6-sulfate. These findings identified the keratan sulfate rich proteoglycans, prevalent at the surface of joint cartilage, as the most common cartilage matrix molecules that are covalently bound to IgG or to serum albumin by disulfide bonds in the articular cartilage of patients with RA or OA.

Topics: Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfates; Humans; Immunoglobulin G; Keratan Sulfate; Osteoarthritis; Serum Albumin

Three fibrocartilages associated with the proximal interphalangeal joint are described--at the attachment of the central slip to bone, within the slip where it passes over the joint, and the volar plate. Material was obtained at surgery following trauma, Dupuytren's disease and rheumatoid arthritis. The fibrocartilages were structurally distinct and immunolabelled differently with monoclonal antibodies to extracellular matrix components. All fibrocartilages from normal and Dupuytren's fingers contained chondroitin and keratan sulphate. Type II collagen was present in all attachment zones, although there was little in rheumatoid fingers. It was also present in the dorsal hood of some normal fingers, but not in pathological specimens or the volar plate. The results show that the fibrocartilages are dynamic tissues whose composition varies according to function and use, and changes in disease.

Topics: Adult; Aged; Aged, 80 and over; Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfates; Collagen; Dupuytren Contracture; Female; Finger Joint; Humans; Immunohistochemistry; Keratan Sulfate; Male; Middle Aged

Chondroitin sulfate was administered orally to six healthy volunteers, six patients with rheumatoid arthritis and six patients with osteoarthritis. Blood was collected at intervals before and after treatment and the glycosaminoglycan concentration was analyzed in serum using a sensitive assay based on the metachromatic reaction with 1,9-dimethylmethylene blue. The glycosaminoglycan concentration in serum before and after ingestion of chondroitin sulfate was statistically unchanged in all of the subjects studied. We suggest that chondroprotection by orally administered chondroitin sulfate is a biologically and pharmacologically unfounded theory. Any possible benefit to osteoarthritic patients after ingestion of chondroitin sulfate should be sought at the gastrointestinal rather than at the plasmatic or articular cartilage level.

Topics: Administration, Oral; Arthritis, Rheumatoid; Artifacts; Chondroitin Sulfates; Digestive System; Glycosaminoglycans; Humans; Methylene Blue; Osteoarthritis

To investigate the correlation between joint disease and the composition of chondroitin sulfate in the joint fluid, unsaturated disaccharide isomers of chondroitin 4-sulfate (delta di-4S) and chondroitin 6-sulfate (delta di-6S) were measured in joint fluids obtained from patients with osteoarthritis (OA), rheumatoid arthritis (RA), or traumatic arthritis (TA).. These pathologic joint fluids were digested with chondroitinase ABC, and the delta di-4S and delta di-6S produced were determined by high performance liquid chromatography combined with fluorometry.. Total content of delta di-4S plus delta di-6S was 71.8 +/- 30.0 nmoles/ml (mean +/- SD) in OA, 55.4 +/- 29.3 nmoles/ml in RA, and 211 +/- 149 nmoles/ml in TA joint fluids. The ratio of delta di-6S to delta di-4S was 3.81 +/- 0.992 in OA, 1.13 +/- 0.527 in RA, and 5.75 +/- 2.46 in TA joint fluids. Differences between groups were statistically significant.. These results strongly suggest that the levels of chondroitin sulfate isomers and the delta di-6S: delta di-4S ratio in joint fluid reflect the proteoglycan metabolism of joint tissues, particularly of articular cartilage; hence, they could be used to diagnose joint diseases and to predict articular cartilage destruction from such joint diseases.

Topics: Aged; Arthritis; Arthritis, Rheumatoid; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Female; Humans; Hyaluronic Acid; Knee Injuries; Knee Joint; Male; Middle Aged; Osteoarthritis; Synovial Fluid

Systemic lupus erythematosus (SLE) is an autoimmune disease which involves the basement membranes of blood vessels in multiple organs. An important component of the microvasculature is vascular heparan sulfate proteoglycan (HSPG). In this study, we investigated the presence in SLE and other immune disease sera of autoantibodies to purified vascular HSPG. Our data demonstrate that antibody to HSPG is found primarily in SLE sera, and not in sera from controls or patients with other immune diseases. The titer of antibody to HSPG correlated with complement depletion in SLE sera. Antibody to HSPG was frequently found in high titer in SLE patients with renal and neurologic involvement. These studies indicate that our assay for antibody to vascular HSPG detects a pathologically relevant autoantibody in SLE sera. The implications of our findings for pathogenesis of vascular autoimmunity are discussed, including the relationship of anti-vascular HSPG antibody to anti-DNA and antiphospholipid antibodies.

Topics: Administration, Topical; Anti-Inflammatory Agents; Antibody Specificity; Arthritis, Rheumatoid; Basement Membrane; Blotting, Western; Chondroitin Sulfates; Collagen; Complement System Proteins; Dose-Response Relationship, Immunologic; Enzyme-Linked Immunosorbent Assay; Fibronectins; Heparin; Heparitin Sulfate; Humans; Hyaluronic Acid; Laminin; Lupus Erythematosus, Systemic; Muscle, Smooth, Vascular; Polymyalgia Rheumatica; Proteoglycans; Scleroderma, Systemic; Sjogren's Syndrome

The intensity of safranin 'O' staining is directly proportional to the proteoglycan content in normal cartilage. Safranin 'O' has thus been used to demonstrate any changes that occur in articular disease. In this study, staining patterns obtained using monoclonal antibodies against the major components of cartilage proteoglycan chondroitin sulphate (anti CS) and keratan sulphate (anti KS), have been compared with those obtained with safranin 'O' staining, in both normal and arthritic tissues. In cartilage where safranin 'O' staining was not detectable, the monoclonal antibodies revealed the presence of both keratan and chondroitin sulphate. Thus, safranin 'O' is not a sensitive indicator of proteoglycan content in diseases where glycosaminoglaycan loss from cartilage has been severe.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Arthritis; Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfates; Extracellular Matrix; Humans; Immunohistochemistry; Keratan Sulfate; Middle Aged; Osteoarthritis; Phenazines; Proteoglycans

In the cartilage-pannus junction of 14 patients with rheumatoid arthritis (RA) and seven patients with osteoarthritis (OA), monoclonal antibodies to keratan sulphate (KS) and chondroitin sulphate (CS) stained a transitional fibroblastic zone (TFZ) within the pannus in nine RA patients and one OA patient. In three patients this was clearly localised to the cytoplasm of cells in this zone, but in all remaining cases KS and CS could be demonstrated in the surrounding matrix. This area was distinguished from adjacent pannus which contained many blood vessels and cells positive for MHC Class II antigen. Specific markers for glycosaminoglycans have been employed to demonstrate that chondrocyte-derived cells and matrix contribute to the changes seen at the cartilage-pannus junction in RA-affected joints.

Topics: Adult; Aged; alpha 1-Antichymotrypsin; Antibodies, Monoclonal; Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfates; Female; Histocompatibility Antigens Class II; Humans; Keratan Sulfate; Male; Middle Aged; Osteoarthritis

Phospholipase A2 (PLA2) activity has now been identified in rheumatoid synovial fluids. This PLA2 is a calcium-requiring protein of MW 11,000 with a neutral pH optimum. Its activity was inhibited by high concentrations of Mg2+, and by the active site-directed histidine reagent p-bromophenacyl bromide. Ionic and nonionic detergents, or the sulfhydryl reagent dithiothreitol caused loss of enzyme activity. Synovial fluid PLA2 did not interact with sulphated mucopolysaccharides such as heparin or chondroitin sulphate. Release and sequestration of PLA2 in the joint space may contribute to the characteristic rheumatoid inflammatory changes.

Topics: Acetophenones; Arthritis, Rheumatoid; Calcium; Chondroitin Sulfates; Deoxycholic Acid; Dithiothreitol; Heparin; Hot Temperature; Humans; Hydrogen-Ion Concentration; Magnesium; Molecular Weight; Octoxynol; Phospholipases; Phospholipases A; Phospholipases A2; Polyethylene Glycols; Synovial Fluid

Topics: Adult; Arthritis, Rheumatoid; Cell Migration Inhibition; Chondroitin; Chondroitin Sulfates; Female; Humans; Leukocytes; Male; Middle Aged; Osteoarthritis

The formation of soluble complexes between alcian blue dye and sulfated glycosaminoglycans provides the basis for the quantitative spectrophotometric estimation of the total concentration of these polysaccharides. Samples containing microgram quantities of sulfated glycosaminoglycan are mixed with a stable dye solution prepared in 15% phosphoric/2% sulfuric acids and absorbance readings at 480 nm are compared to an appropriate standard curve. The method is rapid, convenient, and reproducible. Analyses are performed under conditions in which there is no interference from the non-sulfated glycosaminoglycan hyaluronic acid, or most other anionic macromolecules. In addition, estimations are not effected by small anions or individual monosaccharides. The method has been used for the determination of the purity of commercially available preparations of hyaluronic acid and for the estimation of the sulfated glycosaminoglycan content of various biological fluids including normal human urine and the synovial fluid of individuals with rheumatoid arthritis and osteoarthritis.

Topics: Alcian Blue; Arthritis, Rheumatoid; Chondroitin Sulfates; Glycosaminoglycans; Humans; Osteoarthritis; Spectrophotometry; Synovial Fluid

From 16 synovial membranes patients with rheumatoid arthritis morphological parameters as well as the GAG distribution pattern have been determined using 2 methods each. The results were checked by discriminant and regression analysis: The total GAG concentration of the synovialis was significantly correlated with the sum of the parameters "fibrin in organization", "granulocytes" "nad "necroses" (acute activity). A highly significant correlation between the age of patients and the severity of chronic morphological alterations (basic activity, especially proliferation) was found. The percentage of CS plus DS was positively correlated with the age of patients as well as with the basic activity of the disease, especially with the proliferation. - Contrary to this, HA was significantly decreased when related to the "chronic" parameters. No correlation could be demonstrated between the GAG components and criteria of the acute activity. The evaluating system of STIEHL and GEILER is to be preferred for obtaining morphologic-biochemical correlations. Furthermore, a diagram is proposed demonstrating the cumulative progression of rheumatoid arthritis in the synovialis as related to the GAG pattern. Its combination with morphological data (scars, proliferation, exudation) may help to describe the development of the disease more acurately.

Topics: Adolescent; Adult; Age Factors; Arthritis, Rheumatoid; Child; Chondroitin Sulfates; Dermatan Sulfate; Female; Glycosaminoglycans; Humans; Hyaluronic Acid; Male; Middle Aged; Synovial Membrane

Hyaline cartilage was obtained from patients undergoing synovectomy of the knee joint for rheumatoid arthritis. Eroded cartilage from beneath the invading pannus and relatively normal cartilage from the same joint were maintained in organ culture for three days. During the first 48 hours in culture the explants were exposed to 35SO4 in the medium. The equivalent layers of normal and eroded cartilage were analysed for DNA uronic acid and 35SO4 incorporation. There was a decrease in the DNA and uronic acid of the eroded cartilage, although only the latter reached statistical significance. The uptake of radioactive sulphate was significantly greater in explants taken from the eroded site than from normal areas. This increase in metabolic activity could well be a protective phenomenon.

Topics: Adult; Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfates; DNA; Glycosaminoglycans; Humans; Knee Joint; Organ Culture Techniques; Sulfates; Uronic Acids

Topics: Arthritis; Arthritis, Rheumatoid; Chondroitin Sulfates; Female; Fructose-Bisphosphate Aldolase; Humans; Hyaluronic Acid; Immunoglobulins; Leukocyte Count; Male; Prednisolone; Synovial Fluid; Viscosity

Topics: Arthritis; Arthritis, Rheumatoid; Blood Proteins; Chondroitin; Chondroitin Sulfates; Humans; Lipids; Serum Globulins

Topics: Antibodies; Antigens; Arthritis; Arthritis, Rheumatoid; Chondroitin; Chondroitin Sulfates; Humans; Streptococcus