Background:Burn injury have significant morbidity and mortality rates. Tissue around burn injury frequently damaged as a secondary wound (secondary thermal injury). There is no clear consensus of second degree burns treatment, as many variability of wound care materials. Glutathione is one of the antioxidants that can minimize the effects of oxidative stress by increasing intracellular Glutathione and accelerating wound healing by increasing the capacity of fibroblast contraction. Aim: The study seeks to examine the effect of topical Glutathione on the healing process of deep dermal burns using male Wistar rats (Rattus Novergicus) as the experimental subjects. Material and Methods: This research is a true experimental study using post test only control group design. Thirty-two rats were treated for burns on the backs (dorsum) by attaching heated iron plate at 100°C. The samples were divided into 2 groups of 16 rats in treated group with topical Glutathione application, and 16 rats in control group with topical placebo. The area of epithelialization and microscopic observation of fibroblasts, collagen and neovascularization was carried out on 5th day (D-5) and 12th day (D-12) studies. Results:Obtained in microscopic increased fibroblasts maturation, collagen deposition, neovascularization and macroscopic re-epithelialization in group of rat samples applied by topical Glutathione in D-5 and D-12 studies and obtained significant differences compared with rat groups that were applied topical placebo. Conclusion: Topical Glutathione can accelerate wound healing of deep dermal burns in male rats (Rattus Novergicus) wistar strain.