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Figure 1.
Blocking of OSM+TNFα induced collagen release and collagenolytic activity by growth factors and MMP inhibitors. Bovine nasal cartilage was stimulated with OSM+TNFα in the presence of TGFß1 (10 ng/ml) or IGF1 (200 ng/ml) or BB-94 (10-5 M) or TIMP-1 (100 U/ml) for 14 days. The media were collected and replaced at day 7. The release of collagen into the media, and collagenolytic activity in the media were assayed as described in "Materials and methods". (A) The cumulative collagen release for the 14 day incubation is expressed as a percentage of the total collagen release for each treatment (mean (SD), n=4). (B) Levels of pro- and active collagenolytic activity (expressed as U/ml) are presented for each treatment. The experiments were done in duplicate. Significance was assessed by comparing the release of collagen or collagenolytic activity induced by OSM+TNFα with that for OSM+TNFα +TGFß1 or IGF1, where *p<0.05, **p<0.01, ***p<0.001 using Student's t test.
Figure 2.
Modulation of MMP and TIMP mRNA expression by TGFß1 and IGF1 in OSM+TNFα treated chondrocytes. Primary bovine nasal chondrocytes were cultured in Dulbecco's modification of Eagle's medium (DMEM) containing 10% fetal calf serum to 80% confluence. The medium was changed to serum-free DMEM overnight and then replaced with serum-free DMEM with OSM (5 ng/ml) +TNFα (5 ng/ml) with or without TGFß1 (10 ng/ml) or IGF1 (200 ng/ml) for 24 hours. Total RNA was harvested and analysed by northern blot.