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PubMed2000

Prostaglandin E2 in temporomandibular joint synovial fluid and its relation to pain and inflammatory disorders.

PURPOSE: The aim of this study was to investigate temporomandibular joint (TMJ) synovial fluid (SF) levels of prostaglandin E2 and its relation to general inflammatory activity and its influence on specific TMJ pain in patients with inflammatory TMJ disorders. PATIENTS AND METHODS: The study comprised 24 patients (30 joints) with inflammatory TMJ disorders and 4 healthy persons (6 joints). TMJ pain at rest, tenderness to palpation of the TMJ, and TMJ pressure pain threshold, as well as pain during joint movements (PM), were assessed. PGE2 levels were analyzed in synovial fluid samples (SF-PGE2) and blood plasma (P-PGE2). The erythrocyte sedimentation rate (B-ESR) as well as the serum levels of C-reactive protein (S-CRP) and antinuclear antibodies were determined. RESULTS: PGE2 was undetectable in the plasma and in the TMJ SF of the healthy persons. In the patients, PGE2 was detectable in 20 of the 30 (67%) SF samples. SF-PGE2 was significantly and positively correlated to PM in the patients. There were significant correlations between P-PGE2 and B-ESR as well as the S-CRP. CONCLUSIONS: This study shows that the synovial fluid in patients with TMJ inflammatory disorders frequently has a detectable level of PGE2 that is related to TMJ allodynia. The plasma levels of PGE2 seem to be related to the general inflammatory activity in these patients.

Adult↗
PubMed1997

Separation and characterization of polyethylene wear debris from synovial fluid and tissue samples of revised knee replacements.

A study was made of in vivo-generated polyethylene wear particles as separated from synovial fluid samples and from tissue samples surrounding total knee arthroplasty. A comparison of particle size and morphology between the two particle groups was made to assess any effects of selective tissue capture, and macrophage encapsulation and digestion. In addition, a Raman spectroscopy technique was evaluated that enables positive identification of individual wear particles. The particles of the same size range found in the synovial fluid and tissue samples exhibited a comparable morphology. Notably, submicron-sized debris was present in both the synovial fluid and tissue samples surrounding knees with osteolysis. The novel micro-Raman analysis of individual particles was successful in the categorizing of wear debris as polyethylene or nonpolyethylene.

Aged↗
PubMed1978

Experimental intrinsic healing of flexor tendons based upon synovial fluid nutrition.

The healing process of totally cut and subsequently resutured rabbit flexor tendons kept isolated in the knee joint cavity and free in the synovial fluid was studied by histological and ultrastructural techniques. This experimental model represents a "tissue culture in situ," where the tendon is nourished by diffusion from the synovial fluid only and where no adhesions are formed. Under these conditions there is a proliferation of tendon cells and deposition of collagen resulting in bridging of the suture line. On the basis of these findings, it is assumed that the tendon cells possess an intrinsic potential of repair, provided they obtain a sufficient nutritional supply. In the present experimental model, this nutrition was provided by way of diffusional pathways from the synovial fluid.

Animals↗
PubMed1995

Plasma and synovial fluid kinetics, disposition, and urinary excretion of naproxen in horses.

Naproxen (+6-methoxy-[alpha-methyl]-2-naphthalene acetic acid) is a nonsteroidal anti-inflammatory drug that is used for the treatment of inflammatory conditions in horses. We developed a model that describes the drug's disposition and renal excretion, including synovial fluid disposition and elimination after IV administration in horses. The plasma disposition, after IV administration of 5 mg/kg of body weight, was described by a two-compartment model; mean +/- SD distribution and elimination half-lives were 1.42 +/- 0.42 and 8.26 +/- 2.56 hours, respectively. Plasma concentration of naproxen after IV administration of 5 mg/kg was 55.3 +/- 13.5 and 0.61 +/- 0.42 mg/L at 5 minutes and 48 hours after its administration, respectively. Steady-state volume of distribution was 0.163 +/- 0.053 L/kg, and area under the plasma concentration time-curve was 372.1 +/- 128.2 mg/h/L. The peak synovial fluid concentration of 12.68 +/- 12.39 mg/L was measured at 6 hours, and decreased to 0.71 +/- 0.38 mg/L at 36 hours after naproxen administration. The decrease of naproxen concentration in synovial fluid paralleled that in plasma. The appearance half-life of naproxen in synovial fluid was 4.64 hours, and the elimination half-life was 6.73 hours. Total body clearance was 0.015 +/- 0.006 L/h/kg. The percentage of plasma protein binding was 97.0 +/- 2.9% at plasma concentrations between 5 and 100 mg/L. This was significantly (P < 0.05) higher than the percentage of binding at plasma concentrations of 0.5, 1, and 500 mg/L, which was 75.2 +/- 11.8%. Most of the drug was excreted as glucuronidated naproxen and unconjugated desmethylnaproxen.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗
PubMed1990

Some properties of latent collagenase from human synovial fluid.

Latent collagenase has been isolated in pure form from the rheumatoid synovial fluid. The final preparation, activated by trypsin, yielded a collagenase of specific activity 2,227 units/mg. Electrophoresis in sodium dodecyl sulfate polyacrylamide gels revealed a protein doublet of 54 and 50 kDa. Trypsin or HgCl2 activation resulted in disappearance of the doublet and emergence of a new doublet of 47 and 43 kDa. The latent collagenase could also be activated by leucocyte cathepsin G or plasmin. Neither the latent nor the active collagenase from synovial fluid showed any cross-reactivity with the antibodies against leucocyte collagenase. The trypsin activated collagenase degraded collagen type I, II, III giving typical cleavage products but did not degrade type IV and V collagen.

Electrophoresis, Polyacrylamide Gel↗
PubMed2002

Cell-derived microparticles in synovial fluid from inflamed arthritic joints support coagulation exclusively via a factor VII-dependent mechanism.

OBJECTIVE: To determine the cellular origin of synovial microparticles, their procoagulant properties, and their relationship to local hypercoagulation. METHODS: Microparticles in synovial fluid and plasma from patients with rheumatoid arthritis (RA; n = 10) and patients with other forms of arthritis (non-RA; n = 10) and in plasma from healthy subjects (n = 20) were isolated by centrifugation. Microparticles were identified by flow cytometry. The ability of microparticles to support coagulation was determined in normal plasma. Concentrations of prothrombin fragment F(1+2) (by enzyme-linked immunosorbent assay [ELISA]) and thrombin-antithrombin (TAT) complexes (by ELISA) were determined as estimates of the coagulation activation status in vivo. RESULTS: Plasma from patients and healthy controls contained comparable numbers of microparticles, which originated from platelets and erythrocytes. Synovial microparticles from RA patients and non-RA patients originated mainly from monocytes and granulocytes; few originated from platelets and erythrocytes. Synovial microparticles bound less annexin V (which binds to negatively charged phospholipids) than did plasma microparticles, exposed tissue factor, and supported thrombin generation via factor VII. F(1+2) (median 66 nM) and TAT complex (median 710 microg/liter) concentrations were elevated in synovial fluid compared with plasma from the patients (1.6 nM and 7.0 microg/liter, respectively) as well as the controls (1.0 nM and 2.9 microg/liter, respectively). CONCLUSION: Synovial fluid contains high numbers of microparticles derived from leukocytes that are strongly coagulant via the factor VII-dependent pathway. We propose that these microparticles contribute to the local hypercoagulation and fibrin deposition in inflamed joints of patients with RA and other arthritic disorders.

Adult↗
PubMed1985

Comparison of the effects of different anti-inflammatory drugs on synovial fluid prostanoid concentrations in patients with rheumatoid arthritis.

The effects of one-day treatment with nine nonsteroidal anti-inflammatory drugs and prednisolone on human synovial fluid concentrations of prostanoids were studied. The doses were calculated so as to be approximately equipotent according to clinical experience and the recommendations of the manufacturers. Most of the drugs used reduced clearly PGE2 and TxB2 levels in synovial fluid, but only a slight diminution in 6-keto-PGF1 alpha values was found. Carprofen, diclofenac, indomethacin, naproxen and tolfenamic acid reduced significantly the synovial fluid PGE2 concentrations. Diclofenac and indomethacin also reduced significantly the synovial TxB2 concentrations.

6-Ketoprostaglandin F1 alpha↗
PubMed1994

Synovial fluid analysis. A critical reappraisal.

Analysis of synovial fluid (SF) is among the most useful means of evaluating patients with joint complaints. The best reasons to aspirate a joint include suspicion of infection or crystal-induced arthritis, and generally, the risks of the procedure are acceptably low. While there is no consensus regarding which tests should be considered routine for all SF obtained, data support the performance of a SF white blood cell count with differential, gram stain and culture, and examination for crystals by polarizing microscopy. The separation of SF into groups (noninflammatory, inflammatory, or purulent) should not be relied upon diagnostically. Care must be taken in obtaining and handling joint fluid, and caution applied to interpretation of test results, because over-reliance on any individual test may promote misdiagnosis.

Arthritis↗
PubMed2006

Inhibitor-free DNA for real-time PCR analysis of synovial fluid from horses, cattle and pigs.

The potential of five different commercial DNA isolation methods to remove real-time PCR inhibitors from the synovial fluid of horses, cattle and pigs was investigated. All kits with the exception of one included a silica column-based purification of the DNA. With the fifth kit, DNA purification is achieved by removing contaminating macromolecules by a desalting process. We used a recently developed method based on comparison of the real-time PCR signal of an artificial target incorporated into each PCR reaction in the presence of the isolated DNA from the sample, and in control samples containing water instead of isolated DNA. This was followed by statistical analysis of the data. Inhibition and subsequent reduction of the endpoint fluorescence in the real-time PCR reaction was encountered in many cases. Less frequently, the target copy number in the samples was underestimated. However, we found no experimental evidence of a negative influence of the reduced endpoint fluorescence signal on the detection limit of the real-time PCR assay. All kits tested were useful for analyzing pelleted synovial fluid from horses, cattle and pigs. When analyzing non-pelleted synovial fluid, three kits - two based on silica columns and one employing a desalting process - yielded inhibitor-free DNA for real-time PCR analysis.

Animal Diseases↗
PubMed2006

Direct adenovirus-mediated IGF-I gene transduction of synovium induces persisting synovial fluid IGF-I ligand elevations.

Insulin-like growth factor-I (IGF-I) is one of the most influential growth factors in cartilage repair. Maintenance of adequate IGF-I levels after articular repair procedures is complicated by the short biological half-life of IGF-I in vivo. This study investigated the potential for more prolonged IGF-I delivery through direct adenoviral mediated transduction of synovial tissues in the metacarpophalangeal (MCP) joints of horses. The use of a large animal model provided a structurally similar and metabolically relevant corollary to the human knee. The complete IGF-I coding sequence was packaged into an E1-E3 deleted adenovirus-5 vector under cytomegalovirus promoter control (AdIGF-I), and injected at varying total joint doses to the MCP joints of 14 horses. Direct injection of 20 and 50 x 10(10) AdIGF-I resulted in significant elevations of IGF-I in synovial fluid for approximately 21 days. Synovial tissue taken from injected joints at day 35 following injection and compared to tissue taken preinjection from the same joints revealed elevated synoviocyte IGF-I mRNA levels for the highest viral dose by in situ hybridization and real-time PCR techniques. AdIGF-I injections did not result in significant lameness, joint effusion or elevated total protein concentrations in the synovial fluid. Mild mononuclear infiltration of white blood cells was evident in histologic sections of the synovium in the second highest adenoviral IGF-I dose of 20 x 10(10) particles. Cartilage biopsies taken from all injected joints did not reveal any significant changes in proteoglycan levels nor in histological morphology, which included chondrocyte cloning, architecture, cell type or toluidine blue staining, when compared to control joints. Based on these findings, gene transfer of IGF-I to the synovium of joints can result in significant and persistent elevations of IGF-I ligand in synovial fluid with minimal detrimental effects. Direct IGF-I gene therapy may offer a simple approach in treating patients with acute cartilage injury.

Acute Disease↗
PubMed1996

Hydroxyl radical generation by rheumatoid blood and knee joint synovial fluid.

OBJECTIVE: To demonstrate directly that highly reactive hydroxyl radicals (OH.) can be generated in patients with rheumatoid arthritis and contribute to joint damage, and to examine the ability of blood to cause OH. generation. METHODS: The sensitive and specific technique of hydroxylation of aromatic compounds (salicylate and phenylalanine) was used to measure OH.. Synovial fluid and blood from patients with active rheumatoid arthritis were aspirated and immediately added to tubes containing salicylate and phenylalanine as detectors of OH., or to tubes containing saline as a control. Levels of specific products of attack of OH. upon salicylate (2,3- and 2,5-dihydroxybenzoates) and phenylalanine (ortho- and meta-tyrosines) were measured by high performance liquid chromatography. RESULTS: Synovial fluid samples aspirated into saline never contained ortho- or meta-tyrosines or 2,3-dihydroxybenzoate. Of 53 patients examined, synovial fluid and blood from 36 caused formation of ortho- and meta-tyrosines when aspirated into solutions containing phenylalanine. Repeated sampling from three "positive" patients showed consistent evidence of these hydroxylation products. Similarly, of 22 patients examined, synovial fluid and blood from 18 caused formation of 2,3- and 2,5-dihydroxybenzoates when aspirated into salicylate solutions. Further evidence for the role of OH. was provided by inhibition of the hydroxylation by the specific OH. scavengers mannitol and sodium formate. CONCLUSIONS: Aspirated knee joint fluids and blood from rheumatoid arthritis patients can generate OH., consistent with current views on the importance of this radical as a cytotoxic agent in rheumatoid disease. The ability of body fluids to cause OH. formation is not correlated with simple laboratory indices of disease activity, but is reproducible on sequential sampling from the same patients. The mechanism and significance of the phenomenon in rheumatoid arthritis pathology remain to be established.

Arthritis, Rheumatoid↗
PubMed1990

Synovial fluid does not inhibit collagen synthesis. Bovine cruciate ligament studied in vitro.

An organ culture model of four bovine anterior cruciate ligaments was used to compare synovial fluid with bovine serum, lactated Ringer's solution, and Basal Medium Eagle's (BME) to determine their relative abilities to support in vitro conversion of proline into hydroxyproline. There was no difference between serum and synovial fluid in their ability to support collagen synthesis. These results suggest that bovine synovial fluid is not inhibitory in that it is similar to bovine serum in supporting ligament collagen synthesis under the conditions described.

Animals↗
PubMed1993

Activation of neutrophils by soluble and insoluble immunoglobulin aggregates from synovial fluid of patients with rheumatoid arthritis.

OBJECTIVES: Previous work has shown that synovial fluid isolated from patients with active rheumatoid arthritis contains soluble (not sedimented by centrifugation at 11,600 g for two minutes) and insoluble (sedimented by centrifugation at 11,600 g for two minutes) immunoglobulin aggregates that are capable of activating reactive oxidant production by bloodstream neutrophils. The purpose of this study was to determine which of these types of immunoglobulin aggregates activated the secretion of reactive oxygen metabolites and granule enzymes from neutrophils. METHODS: Cell free synovial fluid (from patients with rheumatoid arthritis) was added to neutrophils isolated from blood of healthy controls that had been incubated in the presence and absence of granulocyte-macrophage colony stimulating factor (GM-CSF). Reactive oxidant production was measured by luminol chemiluminescence (which detects both intracellular and extracellular oxidant production) and by cytochrome c reduction (which measures superoxide secretion). RESULTS: The soluble aggregates only activated neutrophils that were previously primed, and activated a rapid and transient burst of reactive oxidant secretion. On the other hand, the insoluble aggregates activated primed and unprimed neutrophils with similar efficacy and most of the oxidants generated (especially in unprimed cells) were intracellular. The soluble aggregates, but not the insoluble aggregates, also activated the secretion of myeloperoxidase from neutrophils that had either been pretreated with cytochalasin B or primed with GM-CSF. CONCLUSION: It is thus proposed that these soluble immunoglobulin aggregates are responsible for activation of the release of tissue damaging granule enzymes and reactive oxidants from primed neutrophils within the rheumatoid joint.

Adult↗
PubMed1991

[Diffusion of imipenem in synovial fluid].

The diffusion of imipenem (IMP) in the knee joint was studied after a 1 g i.v. administration of Tienam over one hour. The synovial fluid was collected under anesthesia during arthroscopy carried out for mechanical lesions of the knee (meniscal lesions after ligamental injuries or sequelae after meniscectomy), in 3 groups of six patients at one, two, or three hours after the end of injection of IMP. The concentrations of IMP determined by high performance liquid chromatography (HPLC) were: 42.5, 20.1, 9.3 and 5.7 mg/l in the blood at T0, T1, T2 and T3 hr, respectively; 20.4, 13.0 and 7.9 mg/l in the synovial fluid at T1, T2 and T3, respectively. The decrease of IMP concentrations in the synovial fluid was 1,5 times as low as in serum. On account of its broad-spectrum antibacterial activity and our data, IMP could be used in perioperative prophylaxis of the knee joint surgery.

Arthroscopy↗
PubMed1993

The quantitation of a native chondroitin sulfate epitope in synovial fluid lavages and articular cartilage from canine experimental osteoarthritis and disuse atrophy.

OBJECTIVE: Previous studies have shown the presence of a native chondroitin sulfate epitope in articular cartilage proteoglycans from canine knee joints with experimental early osteoarthritis (OA), but not in normal cartilage. The objective of this study was to quantitate the native epitope recognized by monoclonal antibody 3-B-3 in synovial fluids and articular cartilage of diseased joints. METHODS: An immunoassay with monoclonal antibody 3-B-3, which recognizes a native chondroitin-6-sulfate structure, was developed and used to analyze synovial fluid lavage material and extracts of articular cartilage from canine knee joints with early experimental OA or with mild disuse atrophy, and from control animals. RESULTS: The concentration of epitope in the OA fluids was elevated 33-35-fold, and in the OA articular cartilage extracts it was elevated > 200-fold, compared with samples from the control group. No significant difference was detected in the levels of 3-B-3 epitope in the synovial fluid lavage material or cartilage extracts from the joints of the disuse group versus the control group. CONCLUSION: The native 3-B-3 epitope in articular cartilage and synovial fluids may be a specific marker of ongoing anabolic events in early degenerative joint disease.

Animals↗
PubMed1985

The immunoreactivity, ligand, and cell binding characteristics of rheumatoid synovial fluid fibronectin.

Fibronectin promotes macrophage adherence and expression of Fc receptors, is chemotactic for fibroblasts, and is an opsonin for fibrin and denatured collagen. These properties suggest a role for fibronectin in the modulation of joint inflammation. Since structural modification of the fibronectin molecule has been shown to result in loss or de novo acquisition of opsonic and chemotactic activity, we determined the functional and immunochemical properties of fibronectin isolated from the inflamed joint. Eighty-six percent of synovial fluids obtained from patients with active rheumatoid arthritis (RA) contained fibronectin fragments, and 39% of the fluids no longer displayed the dimeric form. Compared with native fibronectin, RA peptides were as active in promoting synoviocyte chemotaxis and in glycosaminoglycan binding, but displayed lower affinity for fibrin and gelatin. Although comparable with intact protein in augmenting monocyte attachment to gelatin, the RA synovial fluid peptides did not augment monocyte attachment to fibrin. Analysis of whole synovial fluid and isolated fibronectins by enzyme immunoassay showed that the increased fibronectin immunoreactivity, previously reported in RA synovial fluid, measures intact and nearly intact protein and does not measure extensively degraded fragments.

Animals↗
PubMed1981

Synovial fluid hydroxyproline fractions before and after osmic acid treatment in rheumatoid arthritis.

Synovial fluid total, dialysable and non-dialysable hydroxyproline were determined in patients with Rheumatoid Arthritis before intra-articular osmic acid injection and on days 2 and 4 after this local treatment. On day 2 the increase in dialysable hydroxyproline paralleled the increase in non-dialysable hydroxyproline. Patients with the highest pre-treatment dialysable hydroxyproline levels also had the highest levels of this fraction on day 2 suggesting that articular damage proceeds after osmic acid injection. However, on day 4 dialysable hydroxyproline levels were consistently lower than before the treatment; the same pattern was observed with non-dialysable hydroxyproline, except for all the joints but one, where effusion recurred 6-12 months after osmic acid treatment. This study suggests that the determination of synovial fluid non-dialysable levels 4 days after osmic acid injection may provide a clue to the prediction of recurrent joint effusion and possibly justify a complementary injection either with osmic acid or with a beta-emitting isotope.

Adult↗
PubMed1995

Persistence of parvovirus B19 in synovial fluid and bone marrow.

OBJECTIVES: To determine whether parvovirus B19 (B19) persists in rheumatoid arthritis (RA). METHODS: Polymerase chain reaction (PCR) was used to detect parvovirus B19 genome in the synovial fluid cells or peripheral blood mononuclear cells from 61 patients with early RA; bone marrow from one patient was also studied. The synovium or synovial fluid cells from 28 patients with advanced RA, and synovial fluid cell samples from 18 patients with reactive arthritis (as controls) were studied. Two separate sets of primers and probe were used. RESULTS: Parvovirus B19 specific gene sequences were detected in two patients with early arthritis fulfilling the criteria for RA. CONCLUSION: Parvovirus B19 does not play a significant role in the aetiopathogenesis of RA. However, a few cases of a disease indistinguishable from RA may be triggered by parvovirus B19 infection.

Adult↗