Objective: To examine the effect of local application of eucalyptus oil on wound healing in the skin of the rats, both histologically and by using laser Doppler flowmetry. Methods: Total 30 male rats used in this study were divided into 2 groups, the experimental group: 15 rats were treated locally with eucalyptus oil, the control group: 15 rats were treated locally with distilled water. All animals were subjected to dorsal skin incision (2 cm) the treatment was conducted every day until the animals were sacrificed to determine histologically the epithelial thickness and inflammatory cell counts. The wound healing was assessed clinically, and blood perfusion scan was performed with laser Doppler. Results: The results revealed that the wound size was significantly reduced in the eucalyptus treated rats, as compared with the control. Blood perfusion was significantly increased with eucalyptus group as compared with the controls. Hematoxylin and eosin (HE) staining revealed a significant reduction in the inflammatory reaction and accelerated epithelialization in the eucalyptus group as compared with the control. The result shows a high significant difference (p ≤ 0.01) between experimental and control group regarding all result. Conclusion: The present study demonstrated that the local application of eucalyptus oil promoted remarkable neovascularization and skin regeneration and may lead to novel treatments for skin wounds.