Figure 3.

Fibroblast growth factor 2 increases the expression of cartilage-degrading enzymes by bovine intervertebral disc cells. Nucleus pulposus cells isolated from bovine intervertebral disc were cultured in monolayer in 12-well plates at 8 × 10⁵ cells/cm², and were serum-starved by changing the media to serum-free DMEM/F-12 with antibiotics for 24 hours before treatment. Cells were then treated with 0.1 to 10 ng/ml fibroblast growth factor 2 (FGF2) and 100 ng/ml fibroblast growth factor 18 (FGF18), collected after 24 hours, and the total RNA extracted to perform real-time RT-PCR for (a) MMP-13 gene expression and (c) ADAMTS4 and ADAMTS5 gene expression. (b) Conditioned media was subjected to immunoblotting for the product of pro MMP-13. Error bars represent three different donors in three separate experiments.