Background and objectives: The present research was designed to evaluate the chemical composition of Lavandula stoechas essential oil (EOLS) as well as the in vivo wound-healing property. The chemical composition of EOLS was identified by gas chromatography mass spectrometry. Nineteen compounds of EOLS were reported. Linalool was identified as the major chemical compound (24.87%), followed by linalyl acetate (19.10%). EOLS showed a high content of oxygenated compounds (63.54%). In vivo wound healing activity of the topical cream prepared from EOLS (0.5% w/w) was assessed using a circular excision wound model. The wound area (mm2) in all animal groups was estimated and measured on day 0, 4, 8, 11, and 16. Results: The EOLS formulation cream (0.5% v/w) showed the highest effect on wound models when compared to reference Madecassol® (Asiaticoside). On days 4, 11, and 16, wound contractions were 26.4%, 78%, and 96.3% for the EOLS-treated group, and 8.5%, 64.1%, and 86.1% for the vehicle cream-treated group. Animals treated with EOLS cream showed a significant decrease in the epithelization period, wound area, and scar thickness, whereas the rate of wound contraction significantly increased. This is the first such report to be published. Histological analyses were also consistent with the results of the excision experimental method. Treatment with EOLS cream formulation resulted in decreased inflammation and an increased rate of tissue perfusion and proliferation as well as remodeling, along with re-epithelization. Conclusions: Our results support the use of EOLS in the development of pharmaceuticals for the management of wounds, and/or inflammatory-related diseases. Additional studies are needed to elucidate and explain the exact mechanism of its pharmacological activity.