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PubMedDate not supplied

Application of micropreparative capillary electrophoresis and polymer gel desalting techniques to biological samples illustrated using human synovial fluid.

A component of human synovial fluid (SF) has been separated by micropreparative capillary electrophoresis. The problems associated with application of this technique to a raw body fluid are discussed. Desalting of SF by passage through a capillary formed from polyacrylamide gel is examined and shown to cause loss of hyaluronan polymer as well as low-molecular-mass components of the fluid.

Acrylic Resins↗
PubMed1999

Optimised sample preparation of synovial fluid for detection of Chlamydia trachomatis DNA by polymerase chain reaction.

OBJECTIVE: To optimise sample preparation of synovial fluid for Chlamydia trachomatis (CT) specific polymerase chain reaction (PCR). METHODS: Serial dilutions of purified CT elementary bodies in synovial fluid were prepared. The synovial fluid pellet was processed by eight different methods of sample preparation. Then samples were analysed by CT specific PCR. The sensitivity of PCR was the basis of ranking of the eight different methods. RESULTS: Highest sensitivity was achieved by methods including an additional step of DNA isolation. Additional extraction of protein and polysaccharides by cetyltrimethylammonium bromide (CTAB) increased sensitivity. Addition of hyaluronidase did not increase sensitivity of QIAEX-DNA extraction but was necessary, however, before phenol-chloroform-DNA extraction. CONCLUSIONS: The method of synovial fluid sample preparation significantly influences the sensitivity of subsequent PCR. Additional DNA isolation and extraction of PCR inhibitors by CTAB led to higher sensitivity.

Cetrimonium↗
PubMed2002

Delayed neutrophil apoptosis induced by synovial fluid in rheumatoid arthritis: role of cytokines, estrogens, and adenosine.

The fate of neutrophils at sites of inflammation, where these cells are likely exposed to both anti- and proapoptotic influences, needs to be clarified. To investigate this issue, we studied the survival of neutrophils in the presence of articular fluids from RA joints before and after immune complex activation. Eight of eleven samples of RA synovial fluid studied were found to inhibit spontaneous and immune complex-stimulated neutrophil apoptosis. No relationships were found between GM-CSF and TNF-alpha concentrations measured on each sample of synovial fluid studied and the levels of neutrophil apoptosis detectable in the presence of the same synovial fluid. Furthermore, no activity on neutrophil survival was observed at either physiologic or pharmacologic concentrations of estradiol. On the contrary, the synovial fluid anti-apoptotic activity correlates (r(2) = 0.8818, p < 0.0001) with the adenosine detected at concentrations in each sample ranging from 18.7 to 52.4 microM. Finally, synovial fluids were incapable of interfering with neutrophil activation evaluated as superoxide anion production. Our results suggest that the microenvironment of rheumatoid synovial fluid is a proinflammatory milieu responsible for the in loco persistence of activated and long-surviving neutrophils.

Adenosine↗
PubMed1975

Purification of granulocyte neutral protease from human blood and rheumatoid synovial fluid.

The neutral protease activity of human synovial fluid cells, like that of peripheral blood leucocytes, is located in a granule fraction. It can be solubilised by various agents but only 1 M neutral salts do so without inactivation. Salt-solubilised neutral protease has been purified (300 X) from synovial fluid cells; like preparations obtained in the same way (600 X purified) from peripheral blood leucocytes, it has a broad pH profile of activity (pH 7--10.5) and in this, as well as in substrate specificity and sensitivity to activators and inhibitors, it behaves as a serine-histidine type protease similar to elastase (EC 3.4.21.11). The product showed two major components on polyacrylamide gel electrophoresis. Collagenase or chymotrypsin-like activity were not detected.

Arthritis, Rheumatoid↗
PubMed1987

[Effects of pathological synovial fluids on the metabolism of chondrocytes].

The purpose of this study was to clarify the mechanism of joint destruction in rheumatoid arthritis and osteoarthritis by observing the effects of the pathological synovial fluids on proteoglycan (PG) and collagen metabolism of chondrocytes. When chondrocytes from chick embryo were cultured with pathological synovial fluids, especially RA synovial fluid, biosynthesis of both proteoglycan and collagen of chondrocytes were found to increase in proportion to the amounts of pathological synovial fluids applied to the culture medium. Chondrocytes began to synthesize PG of small molecular size, approximately 70,000 in addition to PG of normal molecular size. It is noteworthy that the PG of small molecular size shows a shortening of the glycosaminoglycan link and core protein on undersulfation. These findings indicate that pathological synovial fluids disturb the biochemical regulation of the articular cartilage matrix by altering both PG and collagen metabolisms of chondrocytes.

Animals↗
PubMed1988

Synovial fluid concentrations of interleukin-1 beta and proteoglycans are inversely related.

Interleukin-1 beta (IL-1 beta) and proteoglycans have been quantified by radioimmunoassay (IL-1 beta) and enzyme linked immunosorbent assay (proteoglycans) in synovial fluids and sera from patients with rheumatoid arthritis (RA) and reactive arthritis. All fluids were also tested for their ability to influence proteoglycan metabolism in a cartilage explant culture system. Synovial fluid IL-1 beta concentrations were inversely related to proteoglycan concentrations in samples from both RA and reactive arthritis patients (P less than 0.002 for all patients). There was no statistically significant relation between immunoreactive IL-1 beta concentration and proteoglycan synthesis or degradation in explants cultured in synovial fluid containing medium. Synovial fluid IL-1 beta concentrations were not related to erythrocyte sedimentation rate or joint total leukocyte count. IL-1 beta was not detectable (limit 250 pg/ml) in any unextracted sera. Although it appears likely that IL-1 beta is involved in the inflammatory and degenerative processes in joint disease, our findings indicate that there is no simple positive relationship between immunoreactive levels of this cytokine in synovial fluid and liberation of proteoglycans from articular cartilage as reflected in synovial fluid proteoglycan concentration.

Adult↗
PubMed1981

[Transformation of connective tissue cells by pathological altered synovial fluid as demonstrated by experimental studies (author's transl)].

Joint cavities and tendon sheaths are fissurations in the mesenchyme. The coating synovial lining cell layer is caused through the environment of the synovial fluid. The pathologically altered synovial fluid initiates the transformation and proliferation of the marginal connective tissue cells. Evidence for this thesis are the synovial transformatory processes of the cutaneous connective tissue in the area of a synovial fistula and the formation of synovial like structures in the subcutaneous fatty tissues of the rat following the application of synovial fluid from patients with rheumatoid arthritis.

Animals↗
PubMedDate not supplied

[Change in phospholipid composition of synovial fluid in children with various types of arthritis].

Phospholipid composition of synovial fluid was studied in patients with reactive, rheumatoid and juvenile chronic forms of arthritis as compared with that of normal synovial fluid. The most pronounced alterations of the synovial fluid phospholipid composition (additional phospholipid fractions, increase in content of lyso-derivatives) were found in the patients with rheumatoid and juvenile chronic arthritis, which appear to occur due to activation of endogenous phospholipases A2, C and lysophospholipase A1.

Adolescent↗
PubMed1983

Concentration of carprofen in the serum and the synovial fluid in rheumatoid arthritis patients.

Concentrations of carprofen in the serum and the synovial fluid were determined in 13 patients with classical or confirmed rheumatoid arthritis. Four hours after the oral administration of 100 mg of carprofen, as one capsule, concentrations in the synovial fluid were found to be between 3.5 micrograms/ml and 9.7 micrograms/ml, with a mean value (+/- SD) of 6.2 +/- 1.4 micrograms/ml, and concentrations in the serum were between 4.7 micrograms/ml and 14.8 micrograms/ml, with a mean value of 9.8 +/- 2.9 micrograms/ml. The carprofen concentrations in synovial fluid varied from 36.3 to 103.0% of those in serum, with a mean value of 65.0 +/- 14.4%. There is a highly significant correlation between carprofen concentrations in the serum and the synovial fluid.

Anti-Inflammatory Agents↗
PubMed2003

Effects of tramadol on synovial fluid concentrations of substance P and interleukin-6 in patients with knee osteoarthritis: comparison with paracetamol.

Both the analgesic drugs tramadol and paracetamol are widely used for the symptomatic therapy of osteoarthritis (OA). The aim of this double-blind, randomised study in patients with knee OA was to compare their effects on synovial fluid concentrations of interleukin (IL)-6 and substance P (SP). Moreover, we evaluated plasma and synovial fluid concentrations of tramadol and its active metabolite (O-desmethyl-tramadol, M1) after oral treatment with this drug. Twenty patients were enrolled. A group of 10 patients received tramadol (50 mg three times a day), and another group of 10 patients were treated with paracetamol (500 mg three times a day) for 7 days. Both drugs significantly reduced the intensity of joint pain. The synovial fluid concentrations of SP were significantly reduced only by the treatment with tramadol. In this group of patients, IL-6 synovial fluid concentrations were slightly, but not significantly, decreased. Paracetamol did not significantly change the synovial fluid concentrations of SP and IL-6. After oral administration, a considerable amount of tramadol was measurable in synovial fluid. Both in plasma and synovial fluid the concentrations of M1 were markedly lower than those of tramadol, with a T/M1 ratio of 14.7+/-4.6 and 9.3+/-3.9, respectively. These data demonstrate that the activity of tramadol may involve the modulation of inflammatory mediators. Moreover, they indicate that after oral treatment with tramadol, both the parent drug and its active metabolite can penetrate into synovial fluid.

Acetaminophen↗
PubMed2002

The influence of the acetabular labrum seal, intact articular superficial zone and synovial fluid thixotropy on squeeze-film lubrication of a spherical synovial joint.

A model of synovial fluid (SF) filtration by articular cartilage (AC) in a step-loaded spherical synovial joint at rest is presented. The effects of joint pathology (such as a depleted acetabular labrum, a depleted cartilage superficial zone consistent with early osteoarthritis and an inflammatory SF) on the squeezed synovial film are also investigated. Biphasic mixture models for AC (ideal fluid and elastic porous transversely isotropic two-layer matrix) and for SF (ideal and thixotropic fluids) are applied and the following results are obtained. If the acetabular labrum is able to seal the pressurised SF between the articular surfaces (as in the normal hip joint), the fluid in the synovial film and in the cartilage within the labral ring is homogeneously pressurised. The articular surfaces remain separated by a fluid film for minutes. If the labrum is destroyed or absent and the SF can escape across the contact edge, the fluid pressure is non-homogeneous and with a small jump at the articular surface at the very moment of load application. The ensuing synovial film filtration by porous cartilage is lower for the normal cartilage (with the intact superficial zone) than if this zone is already depleted or rubbed off as in the early stage of primary osteoarthritis. Compared with the inflammatory (Newtonian) SF, the normal (thixotropic) fluid applies favourably in the squeezed film near the contact centre only, yielding a thicker SF film there, but not affecting the minimum thickness in the fluid film profile at a fixed time. For all that, in the unsealed case for both the normal and pathological joint, the macromolecular concentration of the hyaluronic acid-protein complex in the synovial film quickly increases due to the filtration in the greater part of the contact. A stable synovial gel film, thick on the order of 10(-7)m, protecting the articular surfaces from the intimate contact, is formed within a couple of seconds. Boundary lubrication by the synovial gel is established if sliding motion follows until a fresh SF is entrained into the contact. This theoretical prediction is open for experimental verifications.

Acetabulum↗
PubMed1985

Pseudogout presenting with low synovial fluid glucose: identification of crystals by gram stain.

A man developed acute monoarticular ankle arthritis caused by calcium pyrophosphate dihydrate (CPPD) crystals. The clinical syndrome resembled that of a pyogenic arthritis. Synovial fluid analysis revealed a glucose concentration of 13 mg/dL and 99,000 white blood cells/mm3. Only one other report of an extremely low synovial fluid glucose associated with pseudogout could be found. The diagnosis of pseudogout was initially suggested when rhomboidal forms were seen during synovial fluid Gram stain examination. Synovial fluid examination with polarized microscopy was initially negative, but revealed numerous CPPD crystals when repeated on the third hospital day. This case serves to illustrate how pseudogout can mimic pyogenic arthritis in both clinical presentation and low synovial fluid glucose concentration. The examination of Gram-stained synovial fluid can reveal the rhomboidal forms of CPPD crystals. The appearance of these crystals is documented in this report.

Arthritis, Infectious↗
PubMed1977

The isolation and partial characterization of the major glycoprotein (LGP-I) from the articular lubricating fraction from bovine synovial fluid.

The articular lubricating fraction from bovine synovial fluid was prepared by repeated fractionation in three consecutive CsCl density gradients to remove completely traces of hyaluronic acid. The major glycoprotein consituent (LGP-I) was then isolated by repeated gel-permeation chromatography. The yield of the LGP-I component was about 20 mg/litre of synovial fluid. Sedimentation-equilibrium measurements showed that this glycoprotein was homogeneous and the mol.wt. was calculated to be 227500. Amino acids represented 43% (w/w) and carbohydrate constituents 44% (w/w) of the molecule. Threonine, glutamic acid, proline and lysine (224, 127, 242 and 128 residues/1000 residues respectively) were the major amino acids. Galactosamine, galactose and N-acetylneuraminic acid (202, 162 and 114 residues/molecule of LGP-I component respectively) accounted for 98% of the total carbohydrate residues present. Small amounts of mannose and glucosamine (1 and 9mol respectively/mol of LGP-I component) were also present. After treatment of LGP-I component with alkali and NaB3H4 radioactivity was incorporated into alpha-aminobutyric acid and alanine in a molar ratio of 4:1, and radioactive galactosaminitol was isolated by ion-exchange chromatography from a cleaved oligosaccharide fraction. These data demonstrate the presence of threonine and serine -O-GalNAc linkages, but only 25% of the theoretical likages involving threonine were cleaved by a beta-elimination reaction. Digestion of LGP-I component with Pronase followed by chromatography on DEAE-cellulose yielded glycopeptide fractions with a similar amino acid and carbohydrate composition to the intact molecule. Treatment of desialylated and intact LGP-I component with galactose oxidase followed by reduction with NaB3H4 revealed the presence of 52mol of terminal galactose in the intact molecule and 153mol of galactose/mol of LGP-I component after treatment with neuraminidase. The data indicate the LGP-I component is composed of a single polypeptide chain containg more than 150 oligaosaccharide side chains composed of O-GaINAc-Gal distributed over the length of the peptide chain and that terminal sialic acid residues are linked to galactose in two-thirds of these side chains.

Amino Acids↗
PubMedDate not supplied

Current concepts in synovial fluid analysis.

The systematic examination of synovial fluid confirms the noninflammatory nature of degenerative joint disease, is diagnostic of gout, pseudogout, and septic arthritis, and will usually allow the identification of rheumatoid arthritis, systemic lupus erythematosus and Reiter's syndrome.

Cell Count↗
PubMed1978

[Determination of enzyme activities in the synovial fluid of farm animals].

Reported in this paper are methods for spectrophotometric determination of the activities of lactatedehydrogenase (LDH) and aspartate-aminotransferase (AspAT) and for colorimetric determination of the activity of N-acetyl-beta-glucosaminidase (NAG) in the synovial fluid of farm animals. Optimum conditions are suggested, as a result of these studies, for reactions of those enzymes in synovial fluid. The behaviours of these enzymes in synovial fluid were compared to those in the blood serum of domestic animals and man and exhibited close agreement. The test-kits at present commercially available for applications in human medicine and used in the determination of LDH and AspAT activities are suitable also for the determination of enzyme activities in synovial fluid. Enzymes in synovial fluid are highly responsive and, even smallest amounts of synovial fluid, therefore, are sufficient for positive findings in laboratory diagnosis.

Acetylglucosaminidase↗
PubMed1991

Comparison of keratan sulphate concentrations and the size distribution of proteoglycans in the synovial fluid of patients with osteoarthritis and pyrophosphate arthropathy.

In order to evaluate the effect of calcium pyrophosphate dihydrate (CPPD) deposition on articular cartilage catabolism, the proteoglycans released into normal synovial fluid were compared with those in synovial fluid obtained from patients with osteoarthritis (OA), chronic pyrophosphate arthropathy (CPA) and acute pyrophosphate arthropathy (APA). Keratan sulphate (KS) was measured by the modified 1,9-dimethylmethylene blue (DMB) assay in synovial fluids treated with chondroitin ABC lyase. This enzyme was found to eliminate all of the sulphated glycosaminoglycans in synovial fluid except KS. In OA, CPA and APA the concentrations of KS were found to be significantly higher than in normal synovial fluid (NSF) (P less than 0.01). Similar KS concentrations were observed in CPA and APA. In CPA they were significantly higher than in OA (P less than 0.02). The size distribution of proteoglycan fragments varied between different patients with the same disease, but only minor differences were observed in patients with OA and CPA who were matched for age, sex and disease severity. Furthermore, the size distribution of proteoglycan fragments in the acute and chronic phases of pyrophosphate arthritis was similar. Thus although in pyrophosphate arthritis the rate at which proteoglycans are released from the cartilage may be greater than in OA or normal joints, the fundamental processes governing the release of these macromolecules may be the same.

Adult↗
PubMed1986

Ibuprofen kinetics in plasma and synovial fluid of arthritic patients.

After administration of a single dose and at steady state, ibuprofen concentrations were measured simultaneously in plasma and synovial fluid obtained from eight patients with rheumatoid arthritis. By seven hours after a dose at steady state, the mean synovial fluid: plasma ibuprofen concentration ratios were constant, and the synovial fluid levels were, on average, greater than those in plasma. The extent to which ibuprofen was bound to protein was somewhat greater in plasma than in synovial fluid. As a result, the mean synovial fluid:plasma free concentration ratio for seven-hour and later specimens was greater than that based on total concentrations. The degree of accumulation of ibuprofen in each fluid was minimal, consistent with its short half-life.

Adult↗
PubMed1999

Levels of chondroitin sulfate isomers in synovial fluid of patients with hip osteoarthritis.

We determined the levels of chondroitin sulfate (CS) isomers in the synovial fluid of patients with hip osteoarthritis (OA) to investigate their significance as markers reflecting extracellular matrix metabolism in joint tissues. A cross-sectional study of synovial fluid aspirated from 50 hip joints of 50 patients with OA was performed. Concentrations of chondroitin-4-sulfate (C4S) and chondroitin-6-sulfate (C6S) were determined by high-performance liquid chromatography. Levels of each marker molecule were investigated in relation to age and radiological stage of the disease. In all disease stages, the dominant CS isomer in synovial fluid was C6S. There was a significant negative correlation between levels of C6S and age. A significant inverse correlation was also observed between the ratio of C6S to C4S and age. Results of analysis of covariance in which age was covariate showed that the ratio of C6S to C4S in advanced and terminal stage OA was lower than that in early stage OA. The present results indicate that the ratio of CS isomers in synovial fluid in hip OA varies with the severity of disease. These molecules in synovial fluid may serve as a useful marker reflecting extracellular matrix metabolism in OA.

Adult↗