Objectives: This study has been undertaken to investigate the antioxidant activity of the gel formulation from Pistacia atlantica oil extraction on enzymatic antioxidants in experimental wound created in rats. Materials and Methods: Under anesthesia, a square-shaped skin defect (2x2 cm) was created aseptically by surgical incision. Then, animals were randomly allocated to four groups (I- untreated controls, II- topically treated with base gel, III- topically treated with 5% gel, IV- topically treated with 10% gel). Results: The results suggest that topical application of Pistacia atlantica oil gels improved reepithelialization with continuous stratum basalis and a mature granulation tissue and adnexa (hair follicles and sweat gland) compared with control and base gel groups. Treatment with 10% oil gel significantly enhanced the tensile strength, ultimate stress, yield strength and stiffness in this group compared with the control and base gel groups at 21 days post injury. The collagen fibers showed a more organized pattern and the tissue alignment was greater as compared to the control and base gel-treated groups at the same stage. Conclusion: The results suggest that topical application of Pistacia atlantica oil gel improved the morphological, biochemical and biomechanical properties of experimentally-induced wound defects in rats.