Gelatin-hyaluronate sponge with and without antibiotic and epidermal growth factor (EGF) were prepared and compared. Four types of sponges were applied on the full-thickness dorsal skin defect of Wistar rat. The effects of antibiotic and EGF in gelatin-hyaluronate sponge on wound healing were investigated by light microscopy and image analyzer at postoperative days of 5, 12 and 21. An immunohistochemical technique, employing PC10, a monoclonal antibody against proliferating cell nuclear antigen (PCNA) was applied to wounded tissue sections. The number of PC10-positive cells was very high for the sponge with EGF at postoperative day 5, then gradually decreased with time. Also we found that antibiotics restrained the cell proliferation during the migratory phase. The sponge with both antibiotic and EGF showed good wound healing performances on the whole for a healing period. The epithelium was regenerated fast with EGF-impregnated sponges at day 5, but each sample had nearly the same length of regenerated epithelium at day 12.