THE EFFECT OF COLLAGEN SUBSTRATE IN CULTURE MEDIUM ON DNA AND PROTEIN SYNTHESIS OF DERMAL FIBROBLASTS

Rhie, Jong Won; Shim, Hyung Gon; Byeon, Jun Hee; Kwak, Sung Il; Lee, Chong Kun

Abstract

Collagen is the major component of scar tissue. Considerable progress of fibroblast growth kinetics and of collagen synthesis has been achieved in the past decade. We have been interested in fibroblasts activities as they are expressed by cells cultured in collagen substrate. This study is to examine the effects of collagen substrate and peptide growth factors In culture medium on DNA and protein synthesis of human dermal fibroblasts. Collagen, interleukin-1(IL-1) and transforming growth factor-beta(TGF-beta) were added to fibroblast culture media according to the designed experiment model and DNA and protein synthesis were measured by [3H]-thymidine, [3H]-leucine, and [3H]-proline incorporation method. The morphological features of fibroblasts were observed by light microscope. The results were as follows ; 1) There were significant decreases of DNA and protein synthesis of cultured fibroblasts in the presence of collagen substrate compared with those in Control groups(p<0.01). 2) DNA and protein synthesis were decreased as dose dependant manner of collagen density in culture media. 3) Morphological features of fibroblasts became less stellate and flat, more spindle-like in the presence of collagen. 4) In responsiveness to IL-1, collagen non-treated groups responded to IL-1 but collagen treated groups were unresponsive to IL-1 (P<0.05). 5) Cells In collagen non-treated groups responded to TGF-beta as dose-related manner(P<0.01). Collagen treated groups desponded to TGF-beta but did not show TGF-beta dose-dependant relationship. In Conclusion, collagen substrate in the culture medium could lower the DNA and protein synthesis of fibroblasts. Cells in collagen substrate were unresponsive or less responsive to peptide growth factors than those in non-collagen substrate.