The Effect of Ascorbic Acid , TGF - beta , and IFN - gammga on Collagen Synthesis in Three - Dimensional Culture of Normal Human Skin Fibroblast

Kim, Jeong Aee; Chung, Jin Ho; Moon, Sang Eun; Youn, Sang Ho; Kwon, Oh Sang; Eun, Hee Chul

Abstract

BACKGROUND: The classical in vitro method for researching the fibroblast metabolism is a mono layer culture system. But there are a lot of facts that artificial influence of monolayer culture situations do not resemble the in vivo conditions. Recently three-dimensional culture method of fibroblast in collagen gel was developed and in this system fibroblasts seem to behave like in vivo situations.
OBJECTIVE
So we examined the effects of ascorbic acid. TGF-beta, and IFN-gamma on fibroblast in collagen gel culture system and then compared these effects with those in monolayer culture system. We also studied the regulatory mechanism of collagen gene by ascorbic acid, TFG-beta, and IFN-gamma by Northern blot analysis in monolayer culture system.
METHODS
Using fibroblasts of the neonatal foreskin, we measured the collagen synthesis after treatment of ascorbic acid, TGF-b, and IFN-r in three-dimensional culture and monolayer culture system. We aslo checked the change of mRNA of collagen after treatment in monolayer culture.
RESULTS
The relative levels of collagenase-digestible protein synthesis in the three-dimensional culture seemed to be lower than those in the monolayer system. However, difference of fold in crease was not statistically significant. The expression of collagen mRNA was increased after treatment of TGF-beta and decreased after IFN-gamma treatment.
CONCLUSION
From these results, fibroblasts in the three-dimensional culture system might resemble the in vivo conditions The molecular level of regulation of collagen synthesis by TGF-beta. and IFN-gamma was thought to be transcriptional. The three-dimensional culture system could be a good model of culture system of fibroblast in studying of aging process or the role of cytokine in wound healing.