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The basis of the synovial fluid analysis.

Synovial fluid fills the spaces in the joint cavities. Many diseases can affect the joints and frequently only a direct examination of the tissue or synovial fluid will yield the correct diagnosis. The article presents the basis of the synovial fluid analysis and a relevant decision-making scheme. The reliability and applicability of synovial fluid tests are commented upon. The synovial fluid analysis undoubtedly plays an important role in the management of patients with joint diseases. Due to a lack of interlaboratory comparisons, and therefore expert opinions on the interpretation of results, this knowledge and the acquisition of relevant experience should be encouraged to enable evaluation of the clinical applicability of recent discoveries in synovial fluid pathology.

Gentian Violet↗

Epstein-Barr virus infection in peripheral blood mononuclear cells, synovial fluid cells, and synovial membranes of patients with rheumatoid arthritis.

OBJECTIVE: Different infectious agents have been proposed to be involved in the pathogenesis of rheumatoid arthritis (RA). We investigated the role of latent Epstein-Barr virus (EBV) infection in patients with RA. METHODS: Sera of 55 patients with RA and 60 age and sex matched healthy controls were examined for antibodies against EBV encoded antigens (EBNA-1, VCA, and EA) by ELISA. The presence of viral DNA was analyzed by polymerase chain amplification of DNA isolated from peripheral blood mononuclear cells (PBMC) of these samples and from cells of synovial fluid (SF) specimens. Paraffin sections of synovial membranes from 25 patients were subjected to in situ hybridization analysis for the EBV encoded small RNA EBER1 and EBER2. RESULTS: Two-fold increased values of IgG antibodies against EBNA-1 were found in patients with RA in comparison to healthy controls (p = 0.029). No statistically significant difference could be observed for antibody levels against EBV-VCA. Fourteen (24.1%) of 55 patients with RA had serological evidence of reactivated EBV infection in comparison to none of the control group (p = 0.028). In PBMC, EBV DNA was detected in a significantly higher proportion in the patient group (50.9 vs. 30%; p = 0.02). In addition, SF cells harbored the viral DNA in 30% of RA cases compared to 16.6% of control cases (p = 0.02). However, EBER1/2 transcripts could only be found within synovial membranes of 2 (8%) of 25 patients with RA. CONCLUSION: These findings support the hypothesis that EBV infection may be involved in the pathogenesis of RA. Further studies may define the precise pathogenetic mechanisms of viral infection for the development of inflammatory arthritis.

Adolescent↗

The role of synovial fluid filtration by cartilage in lubrication of synovial joints--IV. Squeeze-film lubrication: the central film thickness for normal and inflammatory synovial fluids for axial symmetry under high loading conditions.

The axially symmetric problem of synovial film filtration and synovial gel formation at normal approach of cartilage surfaces in the human hip joint loaded by a compressive force has been solved numerically in Part III of this paper [Hlavácek and Novák, J. Biomechanics 28, 1193-1198 (1995)] for the Newtonian viscous phase of the biphasic synovial fluid and for low loads only. Because of a high non-linearity of the problem the method used there breaks down for higher loads. On anticipating that for a high step loading the fluid pressure in the central part of the squeezed synovial film is close to the pressure in a dry frictionless contact, the synovial film filtration at the film centre is governed by two ordinary differential equations that are easy to solve. The central gel film thickness thus obtained, i.e. the film thickness at the moment, when the filtered fluid turns into a stable gel is about 1 micron for the normal synovial fluid (with a non-Newtonian viscous phase of the synovial fluid) and changes very little if the geometric, material and loading parameters of the problem vary within the physiological range. The inflammatory case (with a more or less Newtonian viscous phase) yields values by one order lower at least. The results of stress analysis in the cartilage for this mixture model suggest the reason for vertical cracking at the free cartilage surface and horizontal splitting at the tide mark observed in osteoarthritic joints.

Biomechanical Phenomena↗

[The clinical significance of synovial fluid examination].

Synovial fluids from patients were examined, including 15 cases of rheumatoid arthritis (RA); 9 cases of sero-negative arthropathy (3 cases of ankylosing spondylitis, 6 cases of reactive arthritis); 1 case of gout arthritis; and 5 cases of osteoarthritis. The synovial fluids contained 330-72600 white blood cells/mm3, 9-97% of which were granulocytes, in RA patients. Other physical features of RA fluids were: straw green color, cloudy or unclear; moderate viscosity; fair or poor mucin clotting; lack of crystals and bacteria. Synthesis of the above may contribute to the diagnosis of RA. We also found that IgG, IgA and IgM of serum and IgG of synovial fluids in 15 cases of RA were higher than in the other arthritis groups; the IgM-RF positive rates of RA synovial fluids and sera were 20% and 35.7% respectively, while all were negative in the other groups. Immunoglobulins and complement levels in the synovial fluids of all the tested arthritis cases were about half of those in the serum.

Arthritis, Rheumatoid↗

Free radical oxidation products in plasma and synovial fluid of horses with synovial inflammation.

Free radical oxidation products, namely conjugated dienes, ultraviolet fluorescence (excitation 325 nm, emission 395 nm) and visible fluorescence (excitation 360 nm, emission 460 nm) were measured in equine synovial fluid exposed to free radicals in vitro and in the plasma and synovial fluids of horses with synovial effusions. The synovial effusions were induced by intra-articularly administered carrageenin (0.3 ml, 1%), which rarely resulted in clinical lameness. The free radicals were generated in vitro by mixtures of iron and ethylene diamine tetra acetate (Fe/EDTA) or mixtures of hypoxanthine and xanthine oxidase (HX/XO). The conjugated diene concentrations and intensity of ultraviolet fluorescence were negligible in plasma and synovial fluid specimens. No increase resulted from incubation of synovial fluids with either a free radical generating system or as a result of the induced inflammation. The intensity of visible fluorescence did not increase in specimens incubated with Fe/EDTA. However, the intensity of visible fluorescence increased in specimens incubated with HX/XO, in synovial effusions induced by carrageenin, in plasma and in synovial fluids aspirated from saline injected controls. The results indicate that the intensity of visible fluorescence of equine synovial fluid increases after exposure to free radicals and during synovitis in the horse, suggesting a possible role for free radicals in the pathogenesis of equine inflammatory joint disease.

Acute Disease↗

Analysis of pirprofen in cerebrospinal fluid, plasma, and synovial fluid by high-performance liquid chromatography with electrochemical detection.

We describe a high-performance liquid chromatographic method, using electrochemical detection, for the determination of pirprofen in cerebrospinal fluid (CSF), plasma, and synovial fluid (SF). A C-18 column with a mobile phase containing acetonitrile acetate:phosphate buffer (pH 3.0) was employed. Samples were added with phosphoric acid, then extracted into dichloromethane, evaporated, and injected into the chromatograph. A detection potential of +0.85 V was applied on the basis of current-potential curves. Good linearity was found for each fluid in the expected range of therapeutic concentrations. The detection limit was 0.1 ng/mL for CSF, and 1 ng/mL for plasma and SF, with a recovery greater than 96% and intraday coefficient of variation less than 5% in all cases. The main advantages of this method include high specificity and sensitivity which allow the analysis of CSF and the use of small volumes of plasma and SF. The application of the method for the analysis of plasma and SF samples and the kinetic profile in CSF are shown.

Anti-Inflammatory Agents, Non-Steroidal↗

Ligase chain reaction in detection of Chlamydia DNA in synovial fluid cells.

Synovial fluid cells from 12 patients with reactive arthritis (ReA) triggered by Chlamydia trachomatis were studied for the presence of Chlamydia DNA using the ligase chain reaction (LCR) LCx (Abbott) and the polymerase chain reaction (PCR) Amplicor (Roche). In addition, peripheral blood leucocytes from 11 of these patients were analysed by LCR. As controls, seven patients with newly diagnosed rheumatoid arthritis (RA) were included. Chlamydia trachomatis DNA was detectable by LCR in samples of synovial fluid cells from 4/12 patients with C. trachomatis-triggered ReA, and in none by PCR. Chlamydia trachomatis DNA was not detectable in the synovial fluid cells of the seven RA patients by either method, neither was C. trachomatis DNA detectable in the peripheral blood leucocytes of the ReA patients (0/11) or controls (0/6) by LCR. The LCR technique may be useful in the demonstration of Chlamydia DNA in synovial fluid cells.

Adolescent↗

A preliminary study of magnetic resonance relaxation times (T1 and T2) in inflammatory and degenerative synovial fluids.

Multiple synovial fluid samples from 21 patients were analysed using standard synovial analysis techniques and by nuclear magnetic resonance spectroscopy. Significant negative correlations were noted between both T1 (P less than 0.01) and T2 (P less than 0.0006) relaxation times and synovial fluid total protein. No differences in T1 or T2 relaxation times were noted in synovial fluid between 16 patients with inflammatory forms of arthritis and five patients with degenerative arthritis. In a single rheumatoid arthritis patient with concurrent staphylococcal arthritis, T1 and T2 relaxation times did not vary between the active phase and the recovery phase. The lack of any significant differences in the measured relaxation times as a function of joint condition suggest that in vivo magnetic resonance measurements of T1 or T2 for joint analysis may not reveal information of either a diagnostic or pathophysiological nature.

Analysis of Variance↗

Methods and errors in measurements of synovial fluid volume in stifles with low volume and high viscosity synovial fluid. An experimental study in goats.

Synovial fluid (SF) volume was calculated using various methods in the stifles of goats, in which the cranial cruciate ligament had been transected on one side. Measurements were performed prior to surgery and again 4, 8, and 18 weeks following surgery, by measuring the dilution of an injected radioactive tracer diluted by the SF. Later, 7 months following surgery, SF volume measurements using simple arthrocentesis were performed on stifles in 9 of the goats, and the SF that could not be aspirated, was calculated using 2 indirect methods simultaneously on identical fluids in 3 of these goats. SF was also collected directly during staged arthrotomy of the stifles in 4 goats. There were conflicting results between methods, but the resulting calculated SF volumes seemed to be larger in the operated stifles compared to the controls for all the methods at about the same degree. The 2 indirect methods used to calculate the fluid remaining in the joints following arthrocentesis gave disparate volume calculations. The experiments revealed sources of error in all methods. Direct methods failed to acquire the total fluid volume, and indirect methods were subject to improper mixing and escape of the injected fluid or synovial fluid or both. It was concluded that none of the methods could be used to measure the "true" volume of SF, if such a concept exists and can be defined. None of the methods were considered reliable to compare volumes in different type of joints containing this type of fluid. It was, however, concluded that all the methods gave indication of increased SF volume present on a relative basis when paired joints were compared.

Animals↗

Vascular permeability factor/endothelial growth factor (VPF/VEGF): accumulation and expression in human synovial fluids and rheumatoid synovial tissue.

Vascular permeability factor (VPF, also known as vascular endothelial growth factor or VEGF), is a potent microvascular permeability enhancing cytokine and a selective mitogen for endothelial cells. It has been implicated in tumor angiogenesis and ascites fluid accumulation. Since development of the destructive synovial pannus in rheumatoid arthritis (RA) is associated with changes in vascular permeability (synovial fluid accumulation), synovial cell hyperplasia, and angiogenesis, we examined synovial fluids (SFs) and joint tissue for the expression and local accumulation of VPF/VEGF. VPF/VEGF was detected in all of 21 synovial fluids examined and when measured by an immunofluorimetric assay, ranged from 6.9 to 180.5 pM. These levels are biologically significant, since < 1 pM VPF/VEGF can elicit responses from its target cells, endothelial cells. Levels of VPF/VEGF were highest in rheumatoid arthritis fluids (n = 10), with a mean value (+/- SEM) of 59.1 +/- 18.0 pM, vs. 21.4 +/- 2.3 pM for 11 SFs from patients with other forms of arthritis (p = 0.042). In situ hybridization studies that were performed on joint tissues from patients with active RA revealed that synovial lining macrophages strongly expressed VPF/VEGF mRNA, and that microvascular endothelial cells of nearby blood vessels strongly expressed mRNA for the VPF/VEGF receptors, flt-1 and KDR. Immunohistochemistry performed on inflamed rheumatoid synovial tissue revealed that the VPF/VEGF peptide was localized to macrophages within inflamed synovium, as well as to microvascular endothelium, its putative target in the tissue. Together, these findings indicate that VPF/VEGF may have an important role in the pathogenesis of RA.

Adult↗

The occurrence of cryoproteins in synovial fluid; the association of a complement-fixing activity in rheumatoid synovial fluid with cold-precipitable protein.

A significant portion of a complement-fixing activity found in the synovial fluid of patients with rheumatoid arthritis, and previously implicated as a possible cause of the low synovial fluid complement levels in these patients behaves as a high solubility cryoprotein. Analysis of rheumatoid synovial fluid cryoproteins has revealed mixed immunoglobulins, bound complement components, fibrinogen, DNA, and rheumatoid factor. Sorbitol density gradient studies on whole synovial fluid before and after removal of this activity has shown that the complement-fixing activity migrates in the 19S and heavier regions and that portion is removed with cryoprecipitation. Cryoproteins found in nonrheumatoid synovial fluid are generally devoid of complement-fixing activity and predominantly contain fibrinogen. DNA and IgG are also present, with IgG occurring significantly less frequently than in rheumatoid cryoproteins. These findings are discussed in relationship to recent studies demonstrating the presence of complement-fixing antibody to denatured DNA in rheumatoid cryoproteins.

Arthritis, Rheumatoid↗

Antioxidant activity of synovial fluid, hyaluronic acid, and two subcomponents of hyaluronic acid. Synovial fluid scavenging effect is enhanced in rheumatoid arthritis patients.

To test the scavenging of reactive oxygen species (ROS), we added synovial fluids from patients with rheumatoid arthritis (RA) and osteoarthritis, as well as hyaluronic acid (HA) and its 2 subcomponents, D-glucuronic acid and N-acetyl-D-glucosamine, to 2 ROS-generating systems, activated neutrophils and xanthine-xanthine oxidase. Synovial fluid from RA patients, HA, and D-glucuronic acid markedly decreased the O2-, H2O2, OH., and chemiluminescence measured in both systems. HA and synovial fluid, which are known to be susceptible to degradation by excessive ROS in RA patients, also seem to play an active role in protecting articular tissues from oxidative damage.

Acetylglucosamine↗

Preliminary study of joint disease in poultry by the analysis of synovial fluid.

Samples of synovial fluid and synovial membrane were obtained from the hock joints of several groups of broilers, including lame birds and two strains of broilers raised on different feeding regimens and given different drug treatments (carprofen or placebo). There were more significant differences between the groups on the basis of the analysis of the synovial fluid samples than the synovial membrane samples. Experimental birds fed ad libitum had the highest median red blood cell counts and median ghost cell counts of all of the groups, but there were no differences between the groups in the thickness of the synovial lining cell layer or the degree of cellular infiltrate in the synovial membrane. The synovial fluid from the broilers and lame birds fed ad libitum was more turbid, suggestive of intra-articular pathology, and the large numbers of heterophils in samples from the lame birds indicated an inflammatory arthropathy. The birds fed ad libitum which were treated with carprofen had more cells in the synovial fluid than the birds given the placebo. A large number of the samples of synovial fluid from the ad libitum-fed broilers contained blood.

Animal Husbandry↗

Stereoselective disposition of etodolac enantiomers in synovial fluid.

The synovial fluid (SF) uptake of the chiral nonsteroidal anti-inflammatory drug, etodolac, was studied in six arthritic patients, 2 hours (n = 1) or 12 hours (n = 5) after a single 200 mg dose of racemate. Marked stereoselectivity was seen in both SF and plasma; concentrations of pharmacologically inactive R-etodolac were up to 10-fold greater than active S-etodolac. Concentrations of S-etodolac were greater in SF than in plasma (SF:plasma ratio = 1.98 +/- 0.8): No such difference was seen for R-etodolac (SF:plasma = 0.91 +/- 0.3). Considerable concentrations of conjugated enantiomers were present in SF. In vitro equilibrium dialysis studies in drug-spiked samples showed that the unbound fraction of both enantiomers in SF was greater than in plasma; both fluids bound R more extensively than S etodolac. Therapeutically active S-etodolac has greater concentrations in synovial fluid than plasma during the post-distributive phase, which may be of possible clinical relevance.

Aged↗

The synovial fluid.

The synovial fluid is readily available for study in all cases of effusion. Whenever a question of diagnosis arises, the fluid should be removed for study. Removal of large effusions gives temporary relief of pain. The fluid removed should be smeared on a slide, stained with hematoxylin and eosin, and plated for bacterial, viral, and fungal cultures. The solid constituents of the fluid can easily be studied in the simplest of laboratories. Most of the constituents of plasma are present in synovial fluids. Quantitation of glucose, protein, and cells and an attempt to identify crystals, cartilage fragments, or even foreign bodies are crucial to therapy and the evolution of the disease.

Adolescent↗