Studies on the mechanism of action of collagenase from Aebroiwbactor iophagus (ACR-59) in wound healing

ACR-59 ointment is effective on various experimental wounds in rats (burn, decubital ulcer, open and cut wound). We studied the mechanism of action of ACR-59 in wound healing.(1) After producing dermal burns (200℃, 15sec contact, φ 10mm, 20-25g pressure) on the dorsal skin of male Wistar rats, we immediately excised the epidermis and dermis in burned skin, and unburned skin as well, using a cylindrical chisel (φ 9mm). These skins were incubated with ACR-59 solution (Tris-HCl buffer, pH 7.2) at 37℃. The decomposition of each skin by ACR-59 was evaluated by hydroxyproline. ACR-59 decomposed the burned skin in a dose-dependent manner with more than 0.03 nkat/ml, but the unburned skin with 30 nkat/ml. (2) Fibroblasts from fetal rat skin (FR) were cultured with Dulbecco's Modified Eagle's Medium containing 10% fetal bovine serum. The addition of ACR-59 slightly promoted the proliferation of cells with 3.125 nkat/ml but inhibited it with more than 25 nkat/ml. These results suggest that ACR-59 contributes to wound healing by specifically decomposing burned skins, namely necrotic tissue.