Influence of development and joint pathology on stromelysin enzyme activity in equine synovial fluid

Abstract

Objective-To investigate the role of stromelysin (MMP-3) activity in synovial fluid (SF) at diVerent stages of development and in common joint disorders in the horse. Methods-Stromelysin activity was determined with a fluorogenic enzyme activity assay in SF of normal joints of fetal, juvenile and adult horses, and in SF of horses suVering from the developmental orthopaedic disease osteochondrosis (OC) or osteoarthritis (OA). Additionally, MMP-3 activity was expressed as a ratio of previously reported general MMP activity in the same SF samples. Results-The levels of active stromelysin were 30-fold to 80-fold higher in SF from fetal horses than in SF from juvenile and mature animals (p<0.001). Juvenile horses (5 and 11 months of age) showed a twofold to threefold higher stromelysin activity than adult horses ( p<0.05). In OC joints, stromelysin activity was not significantly diVerent from the activity in normal, age matched, control joints. In OA joints the activity was about four times higher than in normal joints (p<0.001).The ratio MMP-3 activity/general MMP activity did not change with age in normal, healthy joints. This ratio was more then twofold increased in OA joints compared with normal joints, indicating selective upregulation of gene expression or activation of proMMP-3, or both, in OA pathology. Conclusions-The significantly higher stromelysin activity in young individuals parallels the higher metabolic activity occurring at rapid growth and diVerentiation at early age. In OC, MMP-3 mediated matrix degradation appears to be not different from normal joints. The increased stromelysin activity in OA joints is in agreement with pathological matrix degradation. In these joints MMP-3 activity is selectively increased compared with normal joints. (Ann Rheum Dis 2000;59:155-157) Matrix metalloproteinases (MMPs) have an integral role in connective tissue turnover. Collagenases (MMP-1, -8 and 13) cleave interstitial collagen triple helices. Gelatinases (MMP-2, and -9) act mainly on unwound collagen and gelatin. Stromelysins (MMP-3, -10 and -11) can degrade proteoglycans, collagen types IX, XI and type II N-telopeptides, and participate in the activation of proMMP-1, -8, -9 and -13. Membrane-type MMPs are able to activate proMMP-2.1-3 MMPs are present mainly in inactive form (> 95% of total), which is either the latent proform, or an inhibited complex with tissue inhibitor of metalloproteinases (TIMPs) or alpha-2-macroglobulin.

Regulation of fibronectin and metalloproteinase expression by Wnt signaling in rheumatoid arthritis synoviocytes

“…Several Wnt proteins, including Wnt-1, are required for skeletal tissue morphogenesis during development (15,17,46). Fibronectin and several MMPs, including MMP-3, are essential mediators of these developmental processes (47)(48)(49). Although the effects of Wnt signaling on fibronectin and MMP-3 expression during developmental processes have not been studied in great detail, our experiments suggest that Wnt-1-like signals in mesenchymal fibroblasts may influence both fibronectin and MMP-3 expression in the synovium.…”

Association of metalloproteinases, tissue inhibitors of matrix metalloproteinases, and proteoglycans with development, aging, and osteoarthritis processes in mouse temporomandibular joint

“…Working on experimentally induced TMJ OA in sheep, Miyamoto et al (2002) demonstrated that activated MMP-2 levels correlated with the initial articular cartilage destruction (2-4 weeks) rather than with the progression of OA while pro-MMP-2 was detected in all age groups. Most of the research on the involvement of MMPs in aging and OA in animals was performed on knee joint cartilages (Brama et al 1998(Brama et al , 2000Flannelly et al 2002;Price et al 2002).…”

Matrix metalloproteinase activities and their relationship with collagen remodelling in tendon pathology

“…Upon cleavage, quenching is lost, and an increase in fluorescence is measured proportional to the amount of hydrolysed substrate (Beekman et al, , 1999. We have designed fluorogenic substrates using fluoresceinyDabcyl as the fluorophoreyquencher combination (Beekman et al, 1999); these substrates are approximately 100-200 times more sensitive than previously used substrates in which EDANSyDabcyl acted as the fluorophorey quencher combination (Beekman et al, , 1999Brama et al, 1998Brama et al, , 2000. We have developed formats in which it is possible to determine MMP-1 (collagenase 1) activity, MMP-3 (stromelysin 1) activity or gelatinolytic (MMP-2, MMP-9 and MMP-13) activity.…”