Chios mastic gum (CMG) derived from the Pistacia lentiscus L. var. chia has shown beneficial effects on a wide range of human disorders beside other traditional uses. In this study the essential oil of CMG was isolated by hydrodistillation from Pistacia lentiscus L. var.Chia followed by the identification of its chemical composition. that sample was dominated by α-pinene (29.2%) and Caryophyllene (9.7 %), The oil acute wound healing activity was tested in vivo on excision wound model using albino rats and showed promising results. mastic gum essential oil is much easier to incorporate it in many dosage forms and formulas compared to the resin itself or its total extract. The results of the in vivo study of acute wound healing model on rats revealed the activity of CM oil since the wound was healed by almost 97% by the end of the 20 days compared to about 79% displayed by the untreated group.