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Volume 3 Supplement 2

21st European Workshop for Rheumatology Research

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MMP-1, MMP-3 and MMP-10 are involved in the degradation of cartilage

Rheumatoid arthritis (RA) is characterised by degradation of cartilage and invasion of fibroblast-like synoviocytes (FLS) into adjacent cartilage. Several families of proteinases are involved in the degradation of cartilage, especially the matrix metalloproteinases (MMP's) and cathepsin K. However, it is not known which MMP's are responsible for the degradation of cartilage and the invasiveness of FLS. In this study, the expression of MMP's 1 to 20 and cathepsin K in cultured FLS obtained from joint replacement surgery from RA, osteo-arthritis (OA) and other non-destructive arthropathies are investigated and compared to the invasiveness of the FLS in a matrigel transwell system. In this matrigel transwell system previous studies have shown that FLS from RA patients were significantly more invasive than FLS from patients with OA or other non-destructive arthropathies.

FLS from synovial tissue of 32 RA, 18 OA and 14 patients with other non-destructive arthropathies were obtained from joint replacement surgery. The FLS were grown to confluency and RNA was isolated at passage 1 or 2. cDNA was synthesized using oligo-dT and reverse transcriptase. Expression of MMP's and cathepsin K was investigated using RT-PCR. For MMP's 2, 3, 7-12, 14-17, 19, 20 and cathepsin K RT-PCR was performed with primers for the MMP under investigation and primers for beta-actin in one mix. For MMP-1 and 13 no primers for beta-actin were in the mix. The intensity of the bands were compared and given a number from 0 (no expression) to 3 (intensity more than beta-actin). These numbers were related to the invasiveness (number of cells) in a matrigel transwell system.

FLS that expressed MMP-1, MMP-3 or MMP-10 were significantly more invasive (median number of invasive cells: 3970, 4525, 4998, respectively) than cells that did not express MMP-1, MMP-3 or MMP-10 (1826, P = 0.02; 3081, P = 0.01; 2537, P = 0.01, respectively). Expression of the other MMP's and cathepsin K did not show a significant relationship with invasive growth. Expression of MMP-9 showed a trend with higher expression in more invasive cells (P = 0.09). From this study it can be concluded that a correlation exists between expression of MMP-1, MMP-3 and MMP-10 and invasiveness of FLS in a matrigel transwell system.

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Tolboom, T., Pieterman, E., van der Laan, W. et al. MMP-1, MMP-3 and MMP-10 are involved in the degradation of cartilage. Arthritis Res Ther 3 (Suppl 2), P046 (2001).

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