Collagenase-1 and collagenase-3 synthesis in normal and early experimental osteoarthritic canine cartilage: an immunohistochemical study

Abstract

Objective: The coexistence of different collagenases in cartilage suggests the possibility of specific roles for these enzymes in the degradation of the collagen network in osteoarthritis (OA). We investigated the in situ synthesis and distribution of collagenase- and collegenase-3 in normal and early experimental OA cartilage.

Methods: The OA model was created on 12 mongrel dogs by sectioning the anterior cruciate ligament of the right stifle joint with a stab wound. Dogs were divided into 3 groups of 4 animals each, and sacrificed at 4, 8, and 12 weeks, respectively. A 4th group (n = 4) of unoperated dogs was used as control. Articular cartilage from femoral condyles and tibial plateaus was examined histologically to grade severity of lesions, and immunohistochemical and morphometric analyses were performed to detect the presence of chondrocytes producing collagenase-1 and -3.

Results: In OA dogs, the histologic severity of lesions increased with time, being most severe at 8 and 12 weeks after surgery. In cartilage from OA compared to unoperated dogs, the immunoreactivity was 5-9 times higher (p < 0.0002) for collagenase-1, and 3-6 times higher (p < 0.0002) for collagenase-3, in both femoral condyles and tibial plateaus. Although the cell score increased throughout the cartilage, comparison of the superficial and upper intermediate layers (superficial) with the lower intermediate and deep layers (deep) revealed a significantly higher level for collagenase-1 (p < 0.007) in the superficial layers, contrary to the collagenase-3 data, which indicated a higher level (p < 0.007) in the deep layers. For collagenase- , the cell score increased steadily up to the 12th week, and for collagenase-3, the elevation peaked at 8 weeks. Correlation between the histologic severity and cell score in cartilage specimens from unoperated and OA dogs revealed the highest coefficient for collagenase- at the superficial layers (r = 0.69, p < 0.0001), while for collagenase-3, this was noted at the deep layers (r = 0.65, p < 0.0004).

Conclusion: The number of chondrocytes involved in the synthesis of collagenase-1 and collegenase-3 increases dramatically in the early phase of OA. However, the difference in the topographic distribution of these enzymes, as well as the variation in their correlation pattern, may reflect a different function allocated for each collagenase in the OA cartilage degradation process.