Background: Skin wounds usually a reason of physical pain for patients, also are an economic load for them. It is essential to find out an efficient approach to stimulate skin healing. Materials and Methods: Seventy-two ♂ rats were equally divided into 8 groups; rats were submitted to a surgical damage induction except group (1) as follow;1)Normal intact skin,2)Control,3)γ-rays (8Gy),4) Human amniotic membrane,5) Chitosan, 6)γ-rays+ Human amniotic membrane,7)γ-rays+ Chitosan, 8)γ-rays+ Human amniotic membrane + chitosan. Three rats from each group were decapitated on the 3rd, 7th and 21st day after wound made. Histopathological study, immuno-histochemical staining for epidermal growth factor receptor (EGFR) and biochemical parameters; tissues tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and interleukin-6 (IL-6) in wound area of skin samples were estimated. Results: γ-irradiation of the rats' skin wound delayed the healing process with less re-epithelization and prominent fatty tissues with severe ↑ in the level of the proinflammatory markers. Treatment of rats' irradiated skin wounds with amniotic membrane and/or chitosan improved the healing process, as indicated by pathological, biochemical and EGFR stain results. The repairing in the wound area was the best in the amniotic membrane group compared to other rats' groups. Also, the γ-rays+ amniotic membrane group at all the experimental times was significantly better than all the other groups treated with γ-rays. Conclusions: Treatment of irradiated rats' skin wounds by amniotic membrane and/or chitosan enhanced the healing of wounds. However, amniotic membrane group was the best at all times of the experiment compared to the other groups.