In this study, gelatin based biodegradable scaffolds (GS) containing various amounts of epidermal growth factor (EGF) were prepared and applied to full skin defects created on rats in order to investigate the EGF dose effect on healing process. Scaffolds were prepared by freeze drying technique and stabilized with either glutaraldehyde (GA) or 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC). In vitro cytotoxicity of the prepared scaffolds was assessed by using human skin fibroblast secondary culture (HS2). Scaffolds crosslinked with GA which showed no cytotoxicity in vitro were selected for in vivo applications, loaded with different amounts of EGF (1 mu g, 5 mu g or 10 mu g per cm(3) of scaffold) and were applied onto full skin defects created on rats. Decrease in the wound size and level of reepithelization were examined and the results showed that, all scaffolds, with or without EGF, demonstrated a potential to enhance wound healing. Healing process became faster and better parallel to the dose of EGF present in the scaffolds. Although wounds demonstrated better and faster healing parallel to the EGF content, the dose should be optimized through examining the effects in molecular level before any clinical application.