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Identification of Neisseria gonorrhoeae in synovial fluid using the polymerase chain reaction.

OBJECTIVE: To analyze synovial fluid (SF) for the presence of Neisseria gonorrhoeae DNA using the polymerase chain reaction (PCR). METHODS: We used a modified, nested PCR to detect the presence of N gonorrhoeae DNA in 41 samples of SF obtained from 10 patients with clinical gonococcal arthritis whose SF samples were sterile by culture and from 27 controls, including 11 patients with Reiter's syndrome. Results obtained using this method were compared with those obtained using the GEN-PROBE system, an RNA-DNA hybridization technique. RESULTS: With nested PCR, N gonorrhoeae DNA was detected in 11 of 14 SF samples obtained from patients with culture-negative clinical gonococcal arthritis but in none of the 11 SF samples from Reiter's syndrome patients. The specificity of this technique was 96.4%, with a sensitivity of 78.6%. The rate of false-positive results was 3.6%. The GEN-PROBE technique was unable to detect N gonorrhoeae ribosomal RNA in any of the samples. CONCLUSION: These findings demonstrate the potential utility of the PCR in confirming the clinical diagnosis of gonococcal arthritis as well as providing insight into the pathogenesis of this disorder in patients whose SF are sterile by standard culture techniques. PCR may also prove helpful in differentiating N gonorrhoeae arthritis from acute Reiter's syndrome.

Adolescent↗

Growth factors, insulin-like growth factor-1 and growth hormone, in synovial fluid and serum of patients with rheumatic disorders.

OBJECTIVE: Synovial fluid (SF) plays an important role in joint function. We evaluated the growth factors, insulin-like growth factor-1 (IGF-1) and growth hormone (GH) in SF and serum from patients with osteoarthritis (OA), rheumatoid arthritis (RA), gout, pseudogout and diffuse idiopathic skeletal hyperostosis (DISH). DESIGN: Standard radioimmunoassay techniques were used to measure concurrent levels of IGF-1 and GH. SF samples and serum samples were obtained concomitantly from 27 patients with OA, 22 patients with RA, nine men with gout, 14 patients with pseudogout and eight men with DISH. RESULTS: In the case of IGF-1, a comparison of serum and SF levels shows that SF levels of IGF-1 are lower than serum levels in all groups. Men and women gave similar values. In contrast, in the case of GH, all groups, except males with RA, had higher GH values in SF when compared with serum values. Individual patients with other forms of arthritis demonstrated similar relationships. CONCLUSION: The finding that IGF-1 is present in levels about one-half as great in SF as compared with serum suggests that IGF-1 may be produced in lesser amounts or is utilized by the patient in customary joint function. The finding that GH is present in SF at values twice as high, or more, of serum levels in inflammatory arthritides suggests that GH may play a role in the pathophysiology of arthritic disorders.

Arthritis, Rheumatoid↗

In vitro induction of proinflammatory cytokine secretion by juvenile rheumatoid arthritis synovial fluid immune complexes.

OBJECTIVE: To characterize juvenile rheumatoid arthritis synovial fluid (SF) immune complexes and to examine their interaction with leukocytes. METHODS: SF immunoglobulin-containing fractions were prepared by sequential chromatography on protein A and Sephacryl 300. Fractions were subdivided according to molecular weight, characterized for immunoglobulin and complement content, and incubated with either promonocytic U937 cells or normal human peripheral blood mononuclear cells (PBMC). RESULTS: High molecular weight SF immunoglobulin-containing fractions stimulated the release of interleukin-1beta (IL-1beta) from U937 cells. These same complexes stimulated tumor necrosis factor alpha (TNFalpha), IL-1beta, IL-6, IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF) from PBMC. Lower molecular weight material was less efficient in inducing any of the cytokines. TNFalpha and IL-1beta were the earliest of the messenger RNAs examined to be induced by the high molecular weight complexes. However, the secretion of IL-6, IL-8, and GM-CSF stimulated by the complexes was not completely dependent upon the secretion of IL-1beta. Addition of IL-1 receptor antagonist to the cell cultures reduced GM-CSF and IL-6 production by 40% and IL-8 production by 25% in PBMC. CONCLUSION: SF immunoglobulin fractions contain immune complexes that vary in size, composition, and phlogistic potential. High molecular weight complexes are capable of inducing a spectrum of proinflammatory cytokines, all of which have been implicated in the pathogenesis of rheumatic disease.

Adolescent↗

Development and application of a radioimmunoassay for interleukin-8: detection of interleukin-8 in synovial fluids from patients with inflammatory joint disease.

A sensitive and specific radioimmunoassay for human interleukin-8 (IL-8) was developed using isotopically labeled homogenous natural protein. The detection limit (20% inhibition of 125I-IL-8 binding) was 30 pg/100 microliters; 50% displacement occurred at 140 pg/100 microliters. There was no cross-reactivity with the structurally and functionally related neutrophil-activating peptides 2 and 3 up to 500 ng/100 microliters. The intra- and inter-assay coefficients of variation were 4 and 7%, respectively. In vitro experiments showed that human fibroblasts triggered by interleukin-1, double-stranded RNA or virus release immunoreactive and biologically active IL-8 in a dose- and time-dependent manner. Monocytes produce immunoreactive IL-8 in the 100 ng/ml range when exposed to plant mitogen, bacterial endotoxin, virus or IL-1. Although the radioimmunoassay was more sensitive than the chemotaxis assay (detection limit 0.6 ng/ml versus 10 ng/ml) a correlation between concentrations of immunoreactive IL-8 and neutrophil chemotactic activity in the supernatants from stimulated monocytes and fibroblasts was observed. In synovial fluids from patients with inflammatory joint disease, IL-8 was clearly demonstrable, but there was no correlation between IL-8 levels and general parameters of disease activity (erythrocyte sedimentation rate and serum levels of C-reactive protein). Synovial fluids from patients with rheumatoid arthritis, seropositive for rheumatoid factor, contained significantly higher concentrations of IL-8 than synovial fluids from seronegative rheumatoid arthritis patients and patients with non-rheumatoid arthritis joint inflammation. There was a highly significant correlation between IL-8 levels and serum titers of rheumatoid factor. These findings suggest that the molecular mechanisms underlying joint inflammation may be distinct in different types of arthritis.

Adult↗

[Concentration of IL-18 and PGE2 in synovial fluid in patients with osteoarthritis and its significance].

OBJECTIVE: To detect the concentrations of interleukin-18 (IL-18) and prostaglandin E2(PGE2) in synovial fluid (SF), and to determine the role of IL-18 and PGE2 in osteoarthritis (OA) pathogenesis. METHODS: IL-18 and PGE2 were measured concurrently in synovial fluid samples from 54 patients with knee OA (OA group) and from 9 controls (control group). Quantitative determination of IL-18 was performed by enzyme-linked immunosorbent assay (ELISA). PGE2 was examined by inhibitory enzyme-linked immunosorbent assay. A linear regression between IL-18 and PGE2 was analysed. RESULTS: The concentrations of IL-18 and PGE2 in SF from the OA group were significantly higher than those from the control group (P<0.01). The average value of IL-18 in the control group was (28.768+/-13.575) x 10(-9)ng/L, and (72.303+/-40.130) x 10(-9)ng/L in the OA group (P<0.01); the average value of PGE2 in the control group was (24.697+/-7.814) x 10(-9)ng/L, and (42.302+/-23.818) x 10(-9)ng/L in the OA group (P<0.01). IL-18 was related with PGE2 in a linear curve fashion (the control group: r=0.76, P<0.001; the OA group: r=0.94, P<0.001). CONCLUSION: IL-18 and PGE2 are significantly higher in the OA group than those in the control group, and they might take part in the cartilage degradation in OA pathogenesis. The increase of IL-18 might induce the increase of PGE2, and that might play an important role in OA pathogenesis.

Adult↗

Analysis of T-cell cultures and clones from a patient with classic rheumatoid arthritis--evidence for the existence of autoreactive T-cell clones in blood and synovial fluid.

Using lectin-free IL-2 as the only initial stimulus, bulk cultures and T-cell clones were established from synovial fluid (SFL) and peripheral blood lymphocytes (PBL) of a patient with rheumatoid arthritis (RA). The cloning efficiency of growing bulk cultures was 3%-4% as evaluated by Poisson statistics and was not enhanced by the addition of autologous synovial fluid or serum. The majority of the cloned T cells expressed the OKT8+ phenotype; several clones were OKT4+ and one clone expressed OKT8+ and OKT4+ antigens. None of the cloned T cells exhibited high NK or lectin-dependent cytotoxicity, although bulk cultures had high NK activity. In primed lymphocyte typing responses, bulk cultures and two T-cell clones established from rheumatoid SFL and PBL showed consistent autoreactivity, which we have never before observed with MLC-derived bulk cultures and T cell clones. One of the autoreactive rheumatoid T-cell clones (B25) was found to provide strong helper activity to autologous B cells in the absence of mitogen. Attempts to reveal reactivity of RA-derived T-cell clones to microbial antigens have so far only been successful with Mycoplasma pneumoniae preparations. Careful analysis of this reactivity revealed, however, that Mycoplasma pneumoniae induces a stimulator cell-dependent mitogenic effect rather than an antigen-specific MHC-restricted T-cell proliferation.

Antigens, Bacterial↗

Cytokine concentrations in the synovial fluid and plasma of rheumatoid arthritis patients: correlation with bony erosions.

Cytokines are important protein mediators in inflammatory joint diseases. The synovial fluid and plasma concentrations of interleukin-1 alpha (IL-1 alpha), interleukin-2 (IL-2), tumour necrosis factor-alpha (TNF-alpha), interferon-alpha (IF-alpha) and interferon-gamma (IF-gamma) were measured by RIA and ELISA in 28 rheumatoid arthritis (RA) patients (5 males and 23 females). Ten patients with knee effusions due to other causes (osteoarthritis, psoriasis, gout, rheumatic fever, systemic lupus erythematosus) were also studied. Eight of the RA patients had erosive disease. The synovial fluid IL-1 alpha and IL-2 concentrations were higher in Group 1 (erosive) [IL-1 alpha: 524 pg/ml (SEM: 127), IL-2: 3.28 ng/ml (SEM: 1.0)] than in either Group 2 (non-erosive) [IL-1 alpha: 241 pg/ml (SEM: 24), IL-2: 1.93 ng/ml (SEM: 0.6)] or Group 3 (non-RA) [IL-1 alpha: 267 pg/ml (SEM: 58), IL-2: 0.35 ng/ml (SEM: 0.6)] (p < 0.003 and p < 0.06 respectively). Plasma IL-1 and IL-2 levels were higher in Group 1 [IL-1 alpha: 408 pg/ml (SEM: 107), IL-2: 4.20 ng/ml (SEM: 1.5)] than in Group 2 [IL-1 alpha 150 pg/ml (SEM: 15), IL-2: 2.58 ng/ml (SEM: 0.7)] or Group 3 [IL-1 alpha: 140 pg/ml (SEM: 11), IL-2: 1.93 ng/ml (SEM: 0.3)] (p < 0.01, p < 0.009 respectively). There were no differences in the IFN-alpha, IFN-gamma or TNF-alpha levels between groups. These findings suggest that plasma cytokines levels may reflect synovial levels and that IL-1 alpha may play a significant role in erosive joint disease.

Adolescent↗

[Blood serum and synovial fluid proteins in Reiter's syndrome].

Individual proteins (transferrin, ceruloplasmin, alpha 2-macroglobulin, IgG, IgA and IgM) were examined in blood and synovial fluid of 11 patients with Reiter's syndrome. ARA-1981 was used as a diagnostic criteria. The control group included 40 patients with rheumatoid arthritis and 31 with osteoarthrosis of the knee joint. Statistically significant differences between the concentrations of individual proteins in patients with Reiter's syndrome, rheumatoid arthritis and osteoarthritis were only established in synovial fluid for IgM and for IgM index Csf/CS.

Adult↗

Importance of synovial fluid aspiration when injecting intra-articular corticosteroids.

OBJECTIVE: The aim of this prospective study was to find if a complete synovial fluid aspiration before injecting intra-articular corticosteroids influences the treatment result. METHODS: The study was performed in 147 patients with rheumatoid arthritis (RA). One hundred and ninety one knees with synovitis were randomised to arthrocentesis (n=95) or no arthrocentesis (n = 96) before 20 mg triamcinolone hexacetonide was injected. The duration of effect was followed up for a period of six months. All patients were instructed to contact the rheumatology department if signs and symptoms from the treated knee recurred. If arthritis could be confirmed by a clinical examination a relapse was noted. RESULTS: There was a significant reduction of relapse in the arthrocentesis group (p = 0.001). CONCLUSION: The study shows that aspiration of synovial fluid can reduce the risk for arthritis relapse when treating RA patients with intra-articular corticosteroids. It is concluded that arthrocentesis shall be included in the intra-articular corticosteroid injection procedure.

Administration, Topical↗

Measurement of total and differential white blood cell counts in synovial fluid by means of an automated hematology analyzer.

We developed a rapid and accurate method for quantifying total and differential white blood cell (WBC) counts by pretreating synovial fluid with hyaluronidase and using an automated hematology analyzer. Forty-seven samples of synovial fluid that had been placed in blood-collection tubes containing ethylenediamine- N,N,N',N' -tetraacetic acid as an anticoagulant were treated with hyaluronidase at 37 degrees C for 10 minutes, and then the total and differential WBC counts were determined by means of the automated hematology analyzer. Results were compared with those achieved by traditional manual counting methods. For the automated method, the coefficient of variation values for within-run precision of the WBC count were 5.27%, 3.56%, and 3.01% at 0.54, 1.12, and 2.05 x 10(9) /L, respectively; the run-to-run coefficient of variation values was less than 10.0%. The total and differential WBC counts obtained by this automated method showed good correlation with those obtained by the hemocytometer method ( r = .998; P < .0001; regression formula, y = 0.986 x - 0.072). Bland-Altman plots indicated no significant discrepancy between the methods. Our evaluation supports the use of this automated hematology analyzer method to measure total and differential WBC counts, which should aid clinical diagnosis.

Arthritis, Infectious↗

An investigation of the abnormal metabolic status of synovial fluid from patients with rheumatoid arthritis by high field proton nuclear magnetic resonance spectroscopy.

The 1H Hahn spin-echo NMR profiles of rheumatoid synovial fluids have been investigated and compared with those of matched serum samples. In addition to markedly elevated lactate and diminished glucose concentrations, inflammatory synovial fluids contained (i) substantially lower levels of NMR-detectable chylomicron-and very-low-density-lipoprotein-associated triacylglycerols which appear to have a shortened mean chain-length, and (ii) high concentrations of ketone bodies (predominantly 3-D-hydroxybutyrate), relative to those of corresponding paired serum samples. These observations confirm the abnormal metabolic status of the inflamed rheumatoid joint and provide evidence for an increased utilisation of lipids for fuel therein.

3-Hydroxybutyric Acid↗

Edema-inducing activity of phospholipase A2 purified from human synovial fluid and inhibition by aristolochic acid.

A neutral-active, Ca2+-dependent phospholipase A2 (PLA2) purified 11,000-fold from human synovial fluid (HSF) induced edema when injected into the mouse foot pad. The edema produced by HSF-PLA2 was dose-dependent and was positively correlated with the dose-dependent in vitro expression of PLA2 activity. Maximum edema was achieved within 45 min after the injection and persisted for at least 6 h. Aristolochic acid [8-methoxy-6-nitrophenanthro(3,4-d)-1,3-dioxole-5-carboxylic acid], a major chemical component derived from various species of Aristolochia plant, produced a dose-dependent inhibition of in vitro phospholipid hydrolysis by HSF-PLA2, porcine pancreatic PLA2, snake venom (Naja naja) PLA2, and PLA2 isolated from human platelet. The sensitivity of these PLA2s to inhibition by aristolochic acid varied markedly: HSF-PLA2 greater than N. naja PLA2 greater than human platelet PLA2 greater than porcine pancreatic PLA2. The inhibition of HSF-PLA2 by aristolochic acid was independent of substrate concentration (18-144 microM) and Ca2+ concentration (0.1-4.0 mM). These observations indicate that inhibition of HSF-PLA2 by aristolochic acid may result from direct interaction with the enzyme. When aristolochic acid was mixed with HSF-PLA2 and then injected into the mouse foot pad, edema was inhibited in a dose-dependent manner and was positively correlated with in vitro inhibition of PLA2 activity. Alkylation of HSF-PLA2 with p-bromophenacyl bromide concomitantly inhibited both enzyme and edema-inducing activity. These results clearly demonstrate that the neutral-active, Ca2+-dependent PLA2 isolated from human synovial fluid is proinflammatory and that catalytic activity is positively correlated with in vivo proinflammatory effects.

Acetophenones↗

The effect of growth factors and synovial fluid on chondrogenesis in perichondrium.

Reconstruction of cartilage with perichondrium depends on the chondrogenic property of the perichondrial fibrocytes. The present investigation concerns the conditions for the differentiation of fibrocytes into chondrocytes both in vivo and in vitro. For the in vivo studies specimens of rib and auricular perichondrium from adult rabbits were wrapped round silicon rods which were enclosed in dialysis bags. One was placed in the suprapatellar pouch of the knee joint and one was placed intraperitoneally in each rabbit. After two months the bags were extracted, the perichondrium prepared for microscopic examination, and the chondrogenesis evaluated. In vitro the perichondrium was divided into small pieces and incubated with tissue culture medium. The medium was supplemented with fetal calf serum, together with epidermal growth factor, platelet derived growth factor, synovial fluid, or with human serum albumin (control group). After three weeks the explants were prepared for microscopy. Chondrogenesis was judged by the degree of cellular enlargement, capsule formation, deposition of matrix, and activation of the outer fibrocytic layer. In vivo, good cartilage development was found in all specimens placed in the knee joint but, in those placed intraperitoneally, little if any chondrogenesis was seen. In vitro profound differentiation occurred in all cultures supplemented with epidermal growth factor and platelet derived growth factor. An equivalent differentiation was found in perichondrium that had been incubated with synovial fluid. We conclude that the differentiation of perichondrial fibrocytes is initiated in vitro by growth factors. In addition, we have shown that synovial fluid contains factors that promote and enhance the development of cartilage from perichondrium.

Animals↗

[Inhibition of cardiolipin binding antibodies from synovial fluids of patients with arthritis by endotoxin].

In the presented study was prooven that aCl antibodies cross-reacting with LPS (endotoxin) from arthritic synovial fluids are part of the aPL antibodies pool and correlated with presence in joints bacterial cell wall components like LPS-showing endotoxin presence in synovial fluids and they may be usefull in differential diagnosis of the different kind of arthritis and also in explanation of their aetiology at all--especially in cases of early, undifferentiated arthritis.

Antibodies, Anticardiolipin↗

Descriptions of therapeutic arthrocenthesis and of synovial fluid in a Nahuatl text from prehispanic Mexico.

Paracelsus is considered to have been the first to record the viscid quality of the synovial fluid. However, his contemporary Bernardino de Sahagún, a Franciscan friar who came to Mexico shortly after the Spanish conquest, obtained from elderly Aztec Indians who spoke only Nahuatl the descriptions of therapeutic arthrocentesis and of the viscid nature of the synovial fluid. They compared the fluid from the knee joint to the viscid fluid from the leaves of the nopal cactus (Opuntia sp.). We here record their description and confirm the accuracy of their comparison.

History of Medicine↗

Osmosin: its effect on plasma and synovial fluid kinetics of indomethacin.

'Osmosin' (sodium indomethacin trihydrate, now withdrawn) produced a constant rate of release of indomethacin into the gut. Paired plasma and synovial fluid samples were obtained at regular intervals following a single dose of 'Osmosin' (19 patients) and after continuous daily dosing (15 patients). Indomethacin is rapidly absorbed and plasma concentrations maintained in the range 0.3 to 0.6 micrograms/ml after the first 4 h. Equilibrium between plasma and synovial fluid occurs, with SF/plasma ratios 0.74 to 0.82 12-24 h after a single dose and up to 0.96 thereafter. The indomethacin synovial fluid/plasma profile is changed by Osmosin from the pattern of a short half-life drug to the pattern typical of a long half-life drug. Results from serial samples obtained by use of indwelling cannulae (in vein and knee joint) show close agreement with our single paired sampling technique. Our method may have theoretical disadvantages but it has many practical advantages.

Adult↗

Characteristics of polyethylene wear particles isolated from synovial fluid after mobile-bearing and posterior-stabilized total knee arthroplasties.

The size, shape, and number of polyethylene wear particles found in synovial fluids of patients 1 year after implantation of 22 well-functioning total knee prostheses (11 contemporary mobile-bearing type, 11 posterior-stabilized type) were determined. Polyethylene wear particles were isolated from synovial fluids and examined by scanning electron microscopy. Particle size (equivalent circle diameter) was 0.81 +/- 0.12 microm (mean +/- standard error) in mobile-bearing types and 0.78 +/- 0.08 microm in posterior-stabilized types. Particle shape (aspect ratio) was 1.94 +/- 0.13 in mobile-bearing types and 2.30 +/- 0.22 in posterior-stabilized types. Total numbers of particles were (1.75 +/- 1.02) x 10(8) in mobile-bearing and (1.16 +/- 0.57) x 10(8) in posterior-stabilized types. The differences in these parameters between the two groups were not statistically significant. In the early stages after surgery, contemporary mobile-bearing types were comparable to posterior-stabilized types in terms of polyethylene wear-particle generation. The present results do not support the proposition that has been put forward in the literature; namely, that the contemporary mobile-bearing design has an advantage, in terms of the polyethylene wear rate. These data suggest that the advantage of complete conformity in the femoro-tibial articulating surface of contemporary mobile-bearing design may be offset by wear of the mobile undersurface and slot, apart from the articulating surface.

Aged↗

Assessment of rheumatoid activity based on clinical features and blood and synovial fluid analysis.

Joint inflammation in rheumatoid arthritis has been assessed, and the most useful guides to disease activity were determined by analysis of synovial fluid and blood together with the history of joint disability. The patient's own evaluation of the amount of pain suffered was the most useful clinical assessment. Differential cell count and glucose estimations were the most helpful guides in the synovial fluid, while C-reactive protein in the serum most accurately reflected disease activity. The effects of systemic steroids on these indices were studied, and the differences between seronegative and seropositive patients noted.

Adolescent↗