Effect of type II collagen in chondrocyte response to TGF-beta 1 regulation.

The in vivo role of the extracellular matrix and the manner in which it interfaces with soluble regulators remains unknown. This study reports the modulation by extracellular type II collagen of TGF-beta 1-stimulated DNA synthesis, proteoglycan synthesis, and mRNA expression for alpha 1(II) procollagen and aggrecan core protein in the adult articular chondrocyte. Bovine chondrocytes were isolated and resuspended in alginate beads which contained increasing amounts of type II collagen from 0 to 1.5% (w/v). Cultures were maintained for 7 days in basal, DMEM, TGF-beta 1 (10 ng/ml), or FBS (10%) supplemented medium. DNA and proteoglycan synthesis were determined by radiotracer incorporation. The relative amounts of mRNA were analyzed by Northern blot analysis. Exogenous collagen increased DNA synthesis in all culture conditions beginning at concentrations of 0.75% (w/v). We observed that extracellular type II collagen augments both TGF-beta 1 stimulated increases of aggrecan gene expression up to 400% and alpha 1(II) procollagen gene expression up to 180% in a dose-dependent fashion. This is distinct from cultures which were either basal or FBS supplemented medium which lacked a dose-dependent change in aggrecan gene expression and demonstrated a decrease in alpha 1(II) procollagen gene expression. Exogenous collagen above 0.75% (w/v) increased proteoglycan synthesis significantly in FBS and TGF-beta 1-stimulated cultures but not in basal cultures. We have demonstrated that the alterations in gene expression that occur in response to TGF-beta 1 are modulated by extracellular type II collagen. This modulation is possible through both transcriptional and posttranscriptional regulatory mechanisms.