The purpose of this study is to investigate whether the effects of topically applied petrolatum and glycerin on the barrier repair of acetone-induced skin damage in 6 beagle dogs. To confirm the effects of petrolatum and glycerin on acetone disrupted skin models, we performed to evaluate the characteristics of transepidermal water loss and SC hydration and scanning electron microscopic observations. TEWL and SC hydration measurements were carried out 3, 6, 12, 24, 48h after applying petrolatum and glycerin during recovery from acute disruption. Our results showed that there were some different effects between petrolatum and glycerin on the acetone damaged skin such as barrier function repair process and SC hydration status. The results indicate that the significant improvement could be observed in glycerin apply more than petrolatum after acetone damages, and further study will be required.