The Effect of Hyaluronic Acid on Fibroblast Proliferation in Vitro and Skin Wound Healing in Vivo

Han, Seung Kyu; Kim, Jung Bae; Park, Hyun; Lee, Byung Il; Kim, Woo Kyung

Abstract

It is not determined yet whether hyaluronic acid up- regulates or down-regulates wound healing. This study was designed to define the effect of hyaluronic acid on proliferation of human dermal fibroblasts in vitro and on skin wound closing in vivo. Fibroblasts were isolated from the dermis of adults and cultivated in the presence of either one of 6 concentrations of hyaluronic acid(0, 0.1, 0.2, 0.5, 1.0, 2.0 mg/ml). The fibroblasts were seeded at 2.0 x 10(4) cells/well in Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient including 10% fetal bovine serum with either one of 6 different concentrations of hyaluronic acid in 24-well plates. The cells were incubated for 6 days. All concentrations of hyaluronic acid stimulated the proliferation of fibroblasts. The best proliferation was seen at 0.2 mg/ml of hyaluronic acid concentration(p = 0.01). For in vivo study, 10 white rats were used. A 5 mm round punch was employed to excise skin and subcutaneous tissue at eight sites on the back. After creating 8 open wounds, 8 concentrations(0, 0.1, 0.2, 0.5, 1.0, 2.0, 5.0, 10.0 mg/ml) of hyaluronic acid were applied. The degrees of wound closing were compared the 6th day under light microscope. Low concentration of hyaluronic acid(0 - 2.0 mg/ml) stimulated the wound closing. However, high concentration of hyaluronic acid(5.0 -10.0 mg/ml) delayed the wound closing. The best wound closing was seen at 0.5 mg/ml of hyaluronic acid (p = 0.032). These results demonstrated that hyaluronic acid influenced human dermal fibroblast proliferation and the skin wound closing in rats, and its concentration was critically important factor.