Increased level of heme oxygenase-1 in rheumatoid arthritis synovial fluid

We investigated the expression and localization of heme oxygenase-1 (HO-1) in synovial fluid and synovial tissue, and examined the stimulation of HO-1 production in rheumatoid synovial fibroblasts (RASFs). Synovial fluid samples were obtained from knee joints of 20 rheumatoid arthritis (RA) and 20 osteoarthritis (OA) patients, and concentration of HO-1 and matrix metalloproteinase-3 (MMP-3) were measured by enzyme-linked immunosorbent assay (ELISA). Synovial tissues obtained from RA or OA patients during total knee arthroplasty (TKA) were used for immunohistochemical analysis of HO-1. HO-1 production by RASFs in response to various cytokines was examined by ELISA. HO-1 levels in synovial fluid were higher in the RA group than in the OA group with significant difference (P < 0.001), and correlated with serum C-reactive protein (CRP) level (r = 0.80, P < 0.01) in the RA group. Higher levels of HO-1 were seen in the RA-L group (Larsen grade III–V) than in the RA-E (Larsen grade 0-II) group (P < 0.001). There was weak correlation between the levels of HO-1 protein and MMP-3 in synovial fluid in the RA group (r = 0.31, P < 0.01), while no positive correlation was observed in OA. Positive immunoreaction for HO-1 was observed in cells of synovial tissue including synovial fibroblasts and cells in synovial pannus. HO-1 protein levels in cultured media of RASFs were increased by stimulation by interleukin-1β at 6 h and tumor necrosis factor-alpha at 12 h, but suppressed by interferon-gamma at 12 and 24 h. These results indicated that HO-1 expression in synovial tissue might be stimulated by inflammatory cytokines. The correlation of HO-1 concentration in synovial fluid with serum CRP and MMP-3 in joint fluid indicated that HO-1 might be useful as a marker of joint inflammation in RA patients.     

Comparison of modern marker proteins in serum and synovial fluid in patients with advanced osteoarthrosis and rheumatoid arthritis

Numerous studies have focused on the significance of modern marker proteins in the synovial fluid of the knee joint and in the serum both, for osteoarthritis (OA) and rheumatoid arthritis (RA). The relationship between the serum concentrations and the concentrations in the synovial fluid is still unclear. Synovial fluid and serum samples were obtained from 13 patients with advanced OA and from 8 patients with severe RA and concentrations of MMP-1, MMP-3, MMP-13, TIMP-1, COMP and MIA/CD-RAP were determined. All values were normalized against the total protein concentrations. Serum concentrations of MMP-13 in the RA-group were statistically higher than the synovial values (P<0.05). MMP-13 was the only marker protein that revealed distinct higher levels in the serum than in the synovial fluid. The study design allows only conclusions about advanced stages of RA and OA. Longitudinal investigations may provide further information about the value of MMP-13 as a potential marker to monitor the course of RA and OA.     

Resistin in inflammatory and degenerative rheumatologic diseases

Aims of the study

To assess and compare resistin levels in the serum and synovial fluid of patients with rheumatoid arthritis (RA; an inflammatory rheumatologic disease) and osteoarthritis (OA; a degenerative rheumatologic disease) and to study the relationship between resistin levels and prognostic factors of RA disease progression.

Patients and methods

This study included a total of 50 patients: 25 with RA and 25 with OA. Full case history was documented for all patients and all underwent a thorough clinical examination and laboratory testing. Body mass index (BMI) values were also calculated. Radiographs were made of OA patients’ knees and RA patients’ hands. Disease Activity Score 28 (DAS28) was calculated for RA patients. Serum and synovial fluid samples were obtained from the effused knees of all patients and tested for resistin level.

Results

Serum resistin levels were higher in RA patients than in those with OA (p?<?0.01). Synovial fluid resistin levels were also higher in RA than OA patients (p?<?0.001). While serum resistin levels correlated with Larsen score and total leukocyte count (TLC), synovial fluid resistin levels correlated with rheumatoid factor (RF) and anti-citrullinated protein antibody (ACPA) levels in addition to Larsen score and TLC.

Conclusion

Resistin levels were found to be higher in the serum and synovial fluid of RA patients than in those with OA. This may suggest a role for resistin in inflammatory rheumatologic diseases. The observed statistically significant correlation between synovial fluid resistin levels and RF, ACPA and Larsen score may suggest that high synovial fluid resistin levels can be considered a poor prognostic factor for RA progression. However, further studies employing a larger cohort of patients are needed to confirm the relevance of resistin as a prognostic marker in RA patients.     

Expression of membrane-type matrix metalloproteinases in synovial tissue from patients with rheumatoid arthritis or osteoarthritis

We investigated the expression of membrane-type matrix metalloproteinase (MT-MMP) and matrix metalloproteinase (MMP) mRNAs in synovial tissue from patients with rheumatoid arthritis (RA, n = 5) or osteoarthritis (OA, n = 5) by Northern blot analysis. Northern analysis demonstrated strong expression of MT1-MMP, MT3-MMP, MMP-1, and MMP-3 and weak expression of MT2-MMP and MMP-8 in synovial tissue from patients with RA or OA. MT4-MMP was not detected. No significant difference was shown in the expression of MT-MMP mRNAs between RA and OA. Synovial tissue of RA or OA patients expressed MT-MMPs as well as MMPs. These results indicate that, in addition to MMPs, MT1-MMP, MT3-MMP, and probably MT2-MMP may play a role in the degradation of bone and cartilage matrix in RA and OA. Such information may provide a clue to the development of a novel therapeutic approach targeted on the prevention of joint destruction. Received: April 30, 2000 / Accepted: September 19, 2000     

Osteoprotegerin and the receptor activator of NF-kappa B ligand in the serum and synovial fluid. A comparison of patients with longstanding rheumatoid arthritis and osteoarthritis

We examined OPG and soluble RANKL in the serum (sOPG, sRANKL) and synovial fluid (synOPG, synRANKL) in patients with rheumatoid arthritis (RA) and osteoarthritis (OA). OPG and RANKL were measured in 85 patients (44 with RA, 41 patients with OA) in serum and synovial fluid as well. For measuring of OPG and RANKL ELISA tests were used. The results of OPG and RANKL were compared with clinical and radiological scores. We found a negative correlation for OPG and RANKL in synovial fluids: not only for the whole group of patients (P< 0.003, r=–0.32), but also for the subgroups (RA: P<0.04, r=–0.28, OA: P<0.002, r=–0.54). SRANKL and synRANKL were positively correlated in the whole group (P<0.01, r=0.25) and in the OA group (P<0.02, r=0.35); the RA group was showing a trend (P<0.063, r=0.24), however. Serum OPG was lower in RA, synOPG higher in OA. The difference between the two patient groups was only significant for synOPG (P<0.03, r=0.056), but not for sOPG (P<0.09, r=0.19), sRANKL (P<0.43, r=0.85) or synRANKL (P<0.11, r=0.22). The synOPG:synRANKL ratio was significantly correlated with the Larsen score (P<0.004, r=0.38). Synovial OPG is significantly decreased in rheumatoid joints, whereby synovial RANKL is increased. Lower synOPG could reflect a lower protective effect on bone, thus leading to an earlier and more pronounced bone destruction in RA. However, the effect of different mediators for joint destruction in RA and OA seems not to be important to the pathophysiological changes in the joints. The upregulation of serum OPG might be the result of the inflammation; in contrast, an upregulation of RANKL could not be found in the serum of patients with RA and OA.     

The levels of leukemia inhibitory factor in synovial tissues of patients with rheumatoid arthritis: inflammation and other proinflammatory cytokines

Abstract

To clarify the effect of leukemia inhibitory factor (LIF) on the destruction of rheumatoid arthritis (RA) joints, we investigated the production of LIF and the expression of LIF mRNA in synovial tissues from patients with RA and osteoarthritis (OA). Synovial fluids from RA were used to measure the LIF concentrations using enzyme-linked immunosorbent assay (ELISA). Immunohistochemistory and RT-PCR were used to examine the expression of LIF by synovial cells. LIF mRNA was detected in all cases in RA synovial cells. Although LIF protein was detected only in 20 cases (19%) in RA synovial fluids, LIF concentration in the synovial fluids significantly correlated with the peripheral leukocyte count (P < 0.001) and C-reactive protein (CRP) (P < 0.01). Moreover, levels of IL-1β, IL-6, and IL-8, but not TNF-α, were significantly correlated with LIF in the RA synovial fluids. LIF production was promoted by IL-1β and TNF-α stimulation; in contrast, IL-1 ra and IL-4 were found to markedly decrease LIF production by cultured synovial cells. LIF appeared to be a cytokine produced by RA synovium leading to a proinflammatory secretion profile. Moreover, IL-4 and IL-1 ra may represent attenuated activity for reducing the effect of the destruction of joints by LIF.     

Expression of matrix metalloproteinase 9 (96-kd gelatinase B) in human rheumatoid arthritis

Objective. To determine the expression of matrix metalloproteinase 9/gelatinase B (MMP-9) in synovial fluid (SF), plasma, and synovial tissue from individuals with rheumatoid arthritis (RA), inflammatory arthritis (IA), and osteoarthritis (OA), using specific monoclonal antibody reagents. Methods. Gelatinolytic activity in the SF and plasma of patients with RA, IA, and OA was assessed by gelatin zymography. A mouse monoclonal antiserum, 277.13, which selectively recognizes soluble latent forms of human MMP-9, was used to quantitate MMP-9 levels in patient synovial effusions, plasma, and synovial tissue with a capture sandwich enzyme-linked immunosorbent assay (ELISA). Fifty-one SF samples (31 RA, 9 OA, 11 IA) were analyzed. Immunolocalization of MMP-9 in RA, OA, and normal synovium was investigated using MMP-9–specific antisera. Results. MMP-9 antigen levels in synovial effusions were elevated 67-fold in RA samples compared with OA samples. In addition, although MMP-9 antigen levels in IA synovial effusions were 2.7-fold less than the values in RA samples, they were elevated 34-fold over the values in OA samples. These data indicate an association between increased MMP-9 levels and inflammatory arthritis. A predominant 92-kd gelatinolytic activity (specifically inhibited by EDTA) was evident in RA and IA samples, but no activity was observed in OA samples. Among 86 plasma samples (17 RA, 9 IA, 60 normal controls) analyzed for MMP-9 antigen levels by immunocapture ELISA, MMP-9 antigen levels were elevated 7-fold in RA plasma compared with normal plasma. RA synovial tissue extracts demonstrated elevated levels of MMP-9 antigen compared with OA synovial tissue. MMP-9 immunolocalization studies demonstrated expression in infiltrating leukocytes (neutrophils and macrophages), endothelial cells, and synovial fibroblasts in RA synovium. Conclusion. Latent MMP-9 and/or MMP-9–tissue inhibitor of metalloproteinases 1 (TIMP-1) complexes are elevated in RA and IA SF compared with OA SF. In addition, MMP-9 is increased in RA plasma versus normal control plasma. Synovial tissue levels of MMP-9 antigen are also elevated in RA versus OA. The tissue distribution of MMP-9 within RA synovium is localized to sites of inflammation comprising surface synovial lining cells, endothelium, and leukocytes. Taken together, these observations suggest that connective tissue turnover occurs as a result of excessive MMP activity over TIMP action in the invading pannus, periarticular tissue, or SF. Further studies such as those used in the present investigation will help elucidate the role of a number of different enzymes and inhibitors in the destructive arthropathies.     

Association between hypoxanthine concentration in synovial fluid and joint destruction in patients with rheumatoid arthritis

Abstract

To evaluate the significance of augmented levels of hypoxanthine in synovial fluids in patients with rheumatoid arthritis (RA), the hypoxanthine level in the synovial fluid was investigated in association with joint damage. Concentrations of hypoxanthine, xanthine and uric acid in synovial fluids from knee joints of 45 patients with RA, six patients with gout and five patients with osteoarthritis were determined by high performance liquid chromatography. Relationships between these oxypurines and markers for joint inflammation or Larsen grade of knee joint X-ray film were analyzed. Hypoxanthine levels were significantly elevated in patients with RA and with gout but not in those with osteoarthritis. In RA patients, levels of synovial fluid hypoxanthine were correlated with matrix metalloproteinases MMP-3 ( r=0.510), but not with C-reactive protein nor synovial fluid cytokines. Among various biological factors in synovial fluid (including cytokines and metalloproteinases) only hypoxanthine levels were significantly ( P<0.05) positively correlated with Larsen’s grade of knee joint. In conclusion, augmented levels of synovial hypoxanthine can indicate joint damage in patients with RA and might be a useful marker in a clinical context.     

The levels of leukemia inhibitory factor in synovial tissues of patients with rheumatoid arthritis: inflammation and other proinflammatory cytokines

 To clarify the effect of leukemia inhibitory factor (LIF) on the destruction of rheumatoid arthritis (RA) joints, we investigated the production of LIF and the expression of LIF mRNA in synovial tissues from patients with RA and osteoarthritis (OA). Synovial fluids from RA were used to measure the LIF concentrations using enzyme-linked immunosorbent assay (ELISA). Immunohistochemistory and RT-PCR were used to examine the expression of LIF by synovial cells. LIF mRNA was detected in all cases in RA synovial cells. Although LIF protein was detected only in 20 cases (19%) in RA synovial fluids, LIF concentration in the synovial fluids significantly correlated with the peripheral leukocyte count (P < 0.001) and C-reactive protein (CRP) (P < 0.01). Moreover, levels of IL-1β, IL-6, and IL-8, but not TNF-α, were significantly correlated with LIF in the RA synovial fluids. LIF production was promoted by IL-1β and TNF-α stimulation; in contrast, IL-1 ra and IL-4 were found to markedly decrease LIF production by cultured synovial cells. LIF appeared to be a cytokine produced by RA synovium leading to a proinflammatory secretion profile. Moreover, IL-4 and IL-1 ra may represent attenuated activity for reducing the effect of the destruction of joints by LIF. Received: February 12, 2002 / Accepted: September 6, 2002     

Increased levels of circulating intercellular adhesion molecule 1 in the sera of patients with rheumatoid arthritis

Objectdive. We sought to assess whether circulating levels of intercellular adhesion molecule 1 (ICAM-1) in patients with rheumatoid arthritis (RA) are elevated and correlate with clinical measures of disease activity and whether this ICAM-1 originates from the synovium. Methods. Circulating ICAM-1 (cICAM-1) levels were determined by sandwich enzyme-linked immunosorbent assay of serum from 61 RA, 18 osteoarthritis (OA), and 11 inflammatory arthritis (IA) patients. In addition, paired serum and synovial fluid samples were collected from 17 RA, 9 OA, and 4 IA patients. The stability of cICAM-1 was assessed by overnight incubation at 37°C. Finally, the potential degradative effects of synovial fluid proteases were assessed by incubation of recombinant soluble ICAM-1 with patient synovial fluid. Results. RA sera contained significantly greater (P < 0.001) levels of cICAM-1 compared with RA synovial fluid and compared with sera or synovial fluid from the OA and IA patients. Circulating ICAM-1 levels were unaffected by overnight incubation at 37°C and were unaffected by exposure to RA, OA, or IA synovial fluid. Serum levels of cICAM-1 demonstrated a weak, but significant (P < 0.05) correlation with the joint score and erythrocyte sedimentation rate in 25 RA patients treated with nonsteroidal antiinflammatory drugs. Conclusion. The greatest elevations of cICAM-1 were seen in RA patient sera. In both RA and OA, synovial fluid cICAM-1 levels were consistently lower than serum levels, suggesting that cICAM-1 did not originate in the synovium. Because the production of cICAM-1 can be increased by cytokines (e.g., interleukin-1, tumor necrosis factor α), elevated levels of circulating ICAM-1 in RA may be reflective of systemic exposure to elevated cytokine levels.     

Cartilage oligomeric matrix protein in serum and synovial fluid of rheumatoid arthritis: potential use as a marker for joint cartilage damage

Abstract

This study examined the serum and synovial fluid concentrations of cartilage oligomeric matrix protein (COMP) in relation to the evolution of joint cartilage damage and the requirement for surgery in 125 patients with rheumatoid arthritis (RA). We compared the erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) level, and matrix metalloproteinase-3 (MMP-3) levels with COMP levels determined by specific enzyme-linked immunosorbent assay (ELISA). Patients were divided into three groups: (1) patients with least erosive disease (LES); (2) patients with more erosive disease (MES); and (3) patients with mutilating disease (MUD). In addition, synovial fluid samples were collected from patients undergoing arthroscopic synovectomy of the knee joint (ASS) and total knee arthroplasty (TKA). Serum COMP levels correlated with the ESR (P < 0.0001, r = 0.374, n = 125) and the CRP level (P = 0.0014, r = 0.281, n = 125). COMP levels did not correlate with the MMP-3 level (P = 0.182, r = 0.114, n = 125). The COMP levels of the LES group were significantly lower than those of the MES or MUD groups. Lastly, synovial fluid COMP levels in the TKA group were higher than in the ASS group. Therefore, these findings suggest that serum and synovial fluid COMP levels in patients with RA may reflect cartilage destruction and are correlated with the ESR and the CRP level, which are indicators of the acute-phase response.     

Cartilage oligomeric matrix protein in serum and synovial fluid of rheumatoid arthritis: potential use as a marker for joint cartilage damage

This study examined the serum and synovial fluid concentrations of cartilage oligomeric matrix protein (COMP) in relation to the evolution of joint cartilage damage and the requirement for surgery in 125 patients with rheumatoid arthritis (RA). We compared the erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) level, and matrix metalloproteinase-3 (MMP-3) levels with COMP levels determined by specific enzyme-linked immunosorbent assay (ELISA). Patients were divided into three groups: (1) patients with least erosive disease (LES); (2) patients with more erosive disease (MES); and (3) patients with mutilating disease (MUD). In addition, synovial fluid samples were collected from patients undergoing arthroscopic synovectomy of the knee joint (ASS) and total knee arthroplasty (TKA). Serum COMP levels correlated with the ESR (P < 0.0001, r = 0.374, n = 125) and the CRP level (P = 0.0014, r = 0.281, n = 125). COMP levels did not correlate with the MMP-3 level (P = 0.182, r = 0.114, n = 125). The COMP levels of the LES group were significantly lower than those of the MES or MUD groups. Lastly, synovial fluid COMP levels in the TKA group were higher than in the ASS group. Therefore, these findings suggest that serum and synovial fluid COMP levels in patients with RA may reflect cartilage destruction and are correlated with the ESR and the CRP level, which are indicators of the acute-phase response.     

Association between hypoxanthine concentration in synovial fluid and joint destruction in patients with rheumatoid arthritis

To evaluate the significance of augmented levels of hypoxanthine in synovial fluids in patients with rheumatoid arthritis (RA), the hypoxanthine level in the synovial fluid was investigated in association with joint damage. Concentrations of hypoxanthine, xanthine and uric acid in synovial fluids from knee joints of 45 patients with RA, six patients with gout and five patients with osteoarthritis were determined by high performance liquid chromatography. Relationships between these oxypurines and markers for joint inflammation or Larsen grade of knee joint X-ray film were analyzed. Hypoxanthine levels were significantly elevated in patients with RA and with gout but not in those with osteoarthritis. In RA patients, levels of synovial fluid hypoxanthine were correlated with matrix metalloproteinases MMP-3 (r=0.510), but not with C-reactive protein nor synovial fluid cytokines. Among various biological factors in synovial fluid (including cytokines and metalloproteinases) only hypoxanthine levels were significantly (P<0.05) positively correlated with Larsen’s grade of knee joint. In conclusion, augmented levels of synovial hypoxanthine can indicate joint damage in patients with RA and might be a useful marker in a clinical context.     

Increased levels of stromelysin-1 and tissue inhibitor of metalloproteinases–1 in sera from patients with rheumatoid arthritis

Objective. To evaluate the efficacy of stromelysin-1 (matrix metalloproteinase–3 [MMP-3]) and tissue inhibitor of metalloproteinases–1 (TIMP-1) in serum as markers for joint inflammation in rheumatoid arthritis (RA). Methods. Levels of both macromolecules in sera from 97 healthy controls, 109 patients with RA, and 47 patients with osteoarthritis (OA) were measured by respective 1-step sandwich enzyme immunoassays. In the patients with RA, serum levels of MMP-3 and TIMP-1 were investigated in relation to laboratory and clinical measures of disease activity. In addition, the relationships between serum and synovial fluid (SF) levels in paired samples from individual patients were examined. Results. Serum levels of both MMP-3 and TIMP-1 in RA patients were significantly higher than those in OA patients and in healthy controls (P < 0.001), and were shown to correlate with traditional systemic markers of inflammation including the erythrocyte sedimentation rate and C-reactive protein level, and with the Lansbury articular index. In addition, it was noted that serum levels of MMP-3 correlated with the corresponding values in paired SF samples obtained concurrently from patients with RA (r_s = 0.588, P < 0.001), while such correlations were not found for TIMP-1 levels. Conclusion. Our results support the notion that levels of both MMP-3 and TIMP-1 in RA patient sera are increased in association with inflammation. Furthermore, the level of MMP-3 in serum provides a particularly useful marker of inflammatory activity in the joints of patients with RA.     

Elevation of activated protein C in synovial joints in rheumatoid arthritis and its correlation with matrix metalloproteinase 2

OBJECTIVE: To investigate the involvement of the anticoagulant serine protease activated protein C (APC) in tissue remodeling in rheumatoid arthritis (RA). METHODS: PC/APC, matrix metalloproteinase 2 (MMP-2), and MMP-9 were detected in synovial fluid by Western blotting, and their antigen levels were quantified by enzyme-linked immunosorbent assay in patients with osteoarthritis (OA) or RA. Enzymatic activity of MMP-2 was assayed using a specific fluorogenic substrate. We developed an improved assay to measure APC activity in synovial fluid utilizing a chromogenic substrate following immunoprecipitation with a specific PC/APC antibody. PC/APC and MMP-2 were localized by immunohistochemistry in RA, OA, and normal synovial tissues. RESULTS: Synovial fluid analysis demonstrated that APC is present in both RA and OA synovial fluid, with APC activity being markedly higher in RA (mean +/- SEM 462 +/- 112 ng/ml versus 136 +/- 42 ng/ml; P < 0.02). A correlation (r(2) = 0.61) was found between APC and MMP-2 activity levels in RA patients, but not in OA patients. Immunohistochemical studies of synovial sections showed colocalization of APC and MMP-2 in endothelial and synovial lining cells. Additionally, APC and MMP-2 coimmunoprecipitated with an anti-PC/APC antibody. CONCLUSION: Our results show, for the first time, that APC and MMP-2 are coordinately up-regulated and tightly bound in RA synovial fluid and colocalized in synovia. Their association suggests that APC may modulate MMP-2 activity in RA.     

RNA released from necrotic synovial fluid cells activates rheumatoid arthritis synovial fibroblasts via toll‐like receptor 3

Objective

To assess the expression of Toll‐like receptor 3 (TLR‐3) protein in synovial tissues and cultured synovial fibroblasts obtained from patients with rheumatoid arthritis (RA) and osteoarthritis (OA) and to investigate the consequences of stimulation of cultured synovial fibroblasts with TLR‐3 ligands.

Methods

TLR‐3 expression in synovial tissues was determined by immunohistochemistry and immunofluorescence, and expression in cultured RA synovial fibroblasts (RASFs) was determined by fluorescence‐activated cell sorting and real‐time polymerase chain reaction techniques. TLR‐3 signaling was assessed by incubating RASFs with poly(I‐C), lipopolysaccharide, palmitoyl‐3‐cysteine‐serine‐lysine‐4, or necrotic synovial fluid cells from RA patients in the presence or absence of hydroxychloroquine or Benzonase. Subsequent determination of interferon‐β (IFNβ), CXCL10, CCL5, and interleukin‐6 (IL‐6) protein production in the culture supernatants was performed by enzyme‐linked immunosorbent assays.

Results

TLR‐3 protein expression was found to be higher in RA synovial tissues than in OA synovial tissues. TLR‐3 expression was localized predominantly in the synovial lining, with a majority of the TLR‐3–expressing cells coexpressing fibroblast markers. Stimulation of cultured RASFs with the TLR‐3 ligand poly(I‐C) resulted in the production of high levels of IFNβ, CXCL10, CCL5, and IL‐6 protein. Similarly, coincubation of RASFs with necrotic synovial fluid cells from patients with RA resulted in up‐regulation of these cytokines and chemokines in a TLR‐3–dependent manner.

Conclusion

Our findings demonstrate the expression of TLR‐3 in RA synovial tissue and the activation of RASFs in vitro by the TLR‐3 ligand poly(I‐C) as well as by necrotic RA synovial fluid cells, and indicate that RNA released from necrotic cells might act as an endogenous TLR‐3 ligand for the stimulation of proinflammatory gene expression in RASFs.

     

The significance of altered gelatinase expression in the synovium of patient with arthritic effusions

In this study we quantified the levels of matrix metalloproteinase-2 and 9 (MMP-2 and 9) in effusions and serial synovial cultures of patients with arthritis of the knee in order to investigate the correlations between MMP and cell counts in effusions as well as the possible roles of the synovium. In 49 patients with arthritis of the knee (series I) we examined the cell counts and the amounts of MMP-2 and 9 in 51 effusions. In 20 knee samples of series I of patients who received arthroscopy (series II), we examined the amounts of MMP-2 and 9 in effusions and serial synovial organ cultures. We also compared the gene expressions of MMP-2 and 9 and MT1-MMP in serial synovial cultures using RT-PCR. In series I, significantly more proMMP-9 appeared in effusions from the inflammatory group than in the non-inflammatory and hemorrhagic group (p <0.001). The levels of proMMP-9 correlated with the neutrophil counts in the effusions (p <0.001). In series II synovial cultures, the activities of latent and activated forms of MMP-2 and 9 in lesional areas were all higher than that in paralesional ones (p <0.05). In RT-PCR analysis, MMP-2, -9 and membrane type 1 MMP mRNA levels of lesional areas also showed increased expression. Our data suggest that the analysis of MMP-9 indicates the inflammatory condition of the joints and that additional synovectomy may be beneficial for patients with inflammatory synovitis, compared with non-inflammatory and hemorrhagic arthritis.Abbreviations ECM Extracellular matrix - GA Gouty arthritis - IDV Integrated density value - MMP Matrix metalloproteinases - OA Osteoarthritis - PVNS Pigmented villonodular synovitis - RA Rheumatoid arthritis - SF Synovial fluid - TIMP Tissue inhibitors of metalloproteinases - TKA Total knee arthroplasty     

Serum and synovial fluid interleukin-17 concentrations in rheumatoid arthritis patients: Relation to disease activity,radiographic severity and power Doppler ultrasound

Aim of the workTo investigate serum and synovial fluid levels of IL-17 in rheumatoid arthritis (RA) patients, and its correlation with disease activity and severity.Patients and methods20 RA patients together with 20 primary knee osteoarthritis (KOA) patients and 15 healthy individuals matched for age and sex as control groups were enrolled in this study. Both RA and KOA patients presented with knee effusion. Paired samples of serum and synovial fluid (SF) were collected from RA, OA patients and serum samples from the healthy individuals. RA disease activity was assessed using DAS-28 score and power Doppler ultrasound (PDUS) according to the European League against Rheumatism (EULAR). Radiographic damage was evaluated according to Larsen score.ResultsSerum levels of IL-17 were significantly elevated in RA patients compared to controls (p < 0.001). Also, SF of IL-17 was significantly higher in RA patients compared to OA patients (p < 0.001). In addition, synovial level of IL-17 was significantly higher in RA patient compared to their serum level (p < 0.001). Regarding disease activity grading among RA patients, significant differences (p < 0.05) in mean serum and synovial IL-17 levels were reported being higher in severe active disease. Positive correlations of serum and SF IL 17 levels with PDUS findings and Larsen score were reported.ConclusionSerum and synovial IL-17 levels were significantly elevated in RA patients which clarifies its possible role in RA pathogenesis and correlates positively with disease activity parameters, PDUS findings and Larsen score. Thus targeting IL-17 may provide a promising role in suppressing RA.     

Magnetic resonance imaging-determined synovial membrane and joint effusion volumes in rheumatoid arthritis and osteoarthritis. Comparison with the macroscopic and microscopic appearance of the synovium

Objective. To evaluate the relationship between synovial membrane and joint effusion volumes determined by magnetic resonance imaging (MRI) and macroscopic and microscopic synovial pathologic findings in patients with rheumatoid arthritis (RA) and osteoarthritis (OA). Methods. Synovial biopsies were performed, and macroscopic grades of synovitis assigned, at preselected knee sites during arthroscopy or arthrotomy in 17 knees with RA and 25 with OA. Synovial inflammation and 9 separate tissue characteristics were graded histologically. Synovial membrane and joint effusion volumes were determined by preoperative MRI, enhanced with intravenous gadopentetate dimeglumine. Results. MRI-determined synovial membrane volumes were correlated with the overall histologic assessment of synovial inflammation (Spearman's σ = 0.55, P < 0.001), with fibrin deposition, with subsynovial mononuclear and polymorphonuclear leukocyte infiltration (σ = 0.51-0.59), and less significantly with macroscopic synovitis, vessel proliferation, and granulation tissue formation (σ = 0.40-0.42). No correlation with synovial lining multiplication, perivascular edema, villous formation, or fibrosis was found (σ < 0.30). Conclusion. MRI-determined synovial volumes are correlated with synovial inflammatory activity. Synovial volumes probably mainly reflect the mass of cell-infiltrated, vascularized subsynovial tissue, but may also be influenced by the cumulative synovial proliferative activity. MRI-determined synovial membrane and effusion volumes may be sensitive markers and/or predictors of disease activity and treatment outcome in RA.