Matrix metalloproteinase activity in equine synovial fluid: influence of age, osteoarthritis, and osteochondrosis

Abstract

Objective-To investigate the influence of age, osteoarthritis (OA), and osteochondrosis (OC) on the matrix metalloproteinase (MMP) activity in the synovial fluid (SF) of equine joints. Methods-SF was collected from normal and osteoarthritic metacarpophalangeal joints (normal: 14 adult, 28 juvenile; OA: 22 adult). And from normal and osteochondrotic tarsocrural joints (5 months: 11 normal, 8 OC; 11 months: 7 normal, 6 OC). Subsequently, overall MMP activity was measured. Results-The level of active MMPs was almost twofold higher in SF from juvenile horses (age up to 11 months) than in SF from mature animals (4-30 years; p<0.001). In juvenile horses MMP activity was higher in 5 month old foals than in 11 month old foals (p<0.01). In adult horses MMP activity was independent of age. In OA joints the activity was nearly twice as high as in normal joints (p<0.001). In OC joints MMP activity was not significantly diVerent from normal, age matched, control joints. Conclusions-MMP activity in SF from normal adult joints is not related to age. In juvenile joints MMP activity is significantly higher than activity in joints from adult animals. It is hypothesised that the gradual decrease in MMP activity with increasing age reflects the declining metabolic activity resulting from ceasing growth and the accompanying decrease in cartilage remodelling. The increased MMP activity in osteoarthritic joints most likely reflects matrix destruction. In osteochondrosis MMP mediated matrix degradation appears not to be diVerent from normal joints. (Ann Rheum Dis 1998;57:697-699) Matrix metalloproteinases (MMPs) are essential for normal matrix turnover and play a key part in pathological conditions like osteoarthritis (OA). Matrix turnover in adult articular cartilage is extremely slow. In young individuals metabolism will have to be maintained at a substantially higher level to allow for growth and remodelling.

Metalloproteinases and tumor necrosis factor-alpha activities in synovial fluids of horses: correlation with articular cartilage alterations

“…These results are in agreement with the data from Clegg et al [9] who found a rise in MMP-9 activities in a broad range of aseptic joint diseases. In osteoarthritic joints, Brama et al [3] also found the MMP-9 activity was nearly twice as high as in normal joints, but, in OCD joints, MMPs activity was not significantly different from normal control joints. In humans with rheumatoid arthritis, the concentrations of MMP-2 and 9 in synovial fluids do not seem to be influenced by the amount of residual cartilage [13].…”

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

“…The MMP activity assays were previously shown to be specific for MMP-1, MMP-3 and gelatinolytic activities (MMP-2yMMP-9yMMP-13), respectively (Beekman et al, , 1999Brama et al, 1998;DeGroot et al, 2001) MMPs associated with collagen turnover were extracted from tendon samples using a standardised protocol and normalised against the dry weight of the tendon to enable the comparison of net enzyme activities between specimens; see Fig. 1 and Tables 1 and 2.…”

“…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.…”

“…Recombinant human MMPs (MMP-1, 2 nM; MMP-2, 0.5 nM; MMP-3, 2 nM; MMP-8, 2 nM; MMP-9, 0.5 nM; MMP-13, 1 nM; MMP-14, 1 nM) were incubated with 125 nM TNO003-F, TNO113-F or TNO211-F at 25 8C in 180 ml buffer A w150 mM Tris pH 7.5, 150 mM NaCl, 5 mM CaCl , 2 1 mM ZnCl , 0.01% (vyv) Brij-35x. The catalytic 2 efficiencies (K yK in 10 yMys at 25 8C), determined 3 cat m as described previously , for all available MMPs for TNO003-F are: MMP1s0.2, (Beekman et al, , 1999Brama et al, 1998) as it is hydrolysed at reasonable rates by all MMPs tested, and it is preferentially cleaved by MMPs showing gelatinolytic activity. TNO003-F is most efficiently cleaved by MMP-3.…”

Plasma Concentrations of a Type II Collagen-derived Peptide and Its Nitrated Form in Growing Ardenner Sound Horses and in Horses Suffering from Juvenile Digital Degenerative Osteoarthropathy

“…In juvenile horses, stromelysin or matrix metalloproteinase-3 (MMP-3) activity in synovial fluid was demonstrated to be twofold to threefold higher than in synovial fluid of adult horses (Brama et al, 2000), and higher in 5-month-old than in 11-month-old foals. In adult horses, MMP activity of synovial fluid was independent of age but was nearly twofold higher in osteoarthritis (OA) than in normal joints (Brama et al, 1998). In adult horses, a significant increase of gelatinase (MMP-9) activity was found in synovial fluids of osteoarthritic joints with severe cartilage lesions (Jouglin et al, 2000).…”