Effect of Estrogen on Fibroblast Proliferation and Collagen Synthesis

Won, Chang Hoon; Han, Seung Kyu; Sung, Nak Kwan; Kim, Woo Kyung

Abstract

Estrogen is a sex steroid hormone which is known to be helpful in preventing aging process in various ways. It is frequently used in hormone replacement therapy (HRT). Basically wound healing is likely to have same process on the view of cell proliferation and extracellular matrix formation. However, it is not determined yet whether estrogen up-regulates or down-regulates wound healing. This study was designed to determine the effect of estrogen on proliferation of human dermal fibroblasts and collagen synthesis in vitro which are most important in wound healing. Fibroblasts were isolated from the dermis of adults and cultivated in the presence of either one of 5 concentrations of estrogen(0, 0.01, 0.1, 1.0, 10microgram/ml). The fibroblasts were seeded at 5.0x103cell/well in Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient including 10% fetal bovine serum with either one of 5 different concentration of estrogen in 96-well plates. The cells were incubated for 3 days. For fibroblast proliferation MTT assay method was used. To measure the production of collagen, the collagen type I carboxy- terminal propeptide enzyme immunoassay was carried out. All concentrations of estrogen stimulated the proliferation of fibroblasts. The best proliferation and maximum stimulation of collagen synthesis was seen at supplementation of 1.0microgram/ml of estrogen. The collagen synthesis per cell was also maximal at concentration of 1.0microgram/ml estrogen. These results demonstrate that estrogen influences human dermal fibroblast proliferation and the estrogen concentration is critically important factor in vitro.