Diabetic wounds regularly take a longer time to heal than wounds in healthy animals. Chitosan (CS) interacts with many cellular processes and it elevates the necessarily expression of growth factors in wound healing. The topical utility of zinc was suggested to limit inflammation, enhance re-epithelialization, and lower bacterial growth in chronic wounds. Also, there is evidence that insulin has effects on healing wounds.Therefore, the wound healing potential of insulin injection with CS/Zno nanocomposite membrane in diabetic rats was assessed. Diabetes was induced by single IP injection of Streptozotocin (STZ) at a dose of 50 mg/kg b.wt. after diabetes induction. on the back of each rat, a full thickness excisional wound had been made. Forty eight male rats were divided into Six groups. Group I: normal wound non-treated, Group II: diabetic wound non-treated, Group II: normal wound treated with CS /ZnO membrane, Group IV: diabetic wound treated with CS/ZnO membrane, Group V: diabetic wound and local insulin treatment (2 U/rat per day), Group VI: diabetic wound treated with Chitosan /ZnO membrane and insulin injection. The findings revealed a notable decline in Basic fibroblast growth factor(bFGF), Vascular endothelial growth factor (VEGF), Transforming growth factor β(TGF-β) and α-Smooth muscle actin(α-SMA)in diabetic non-treated wounds. However, treatment with CS/Zno nanocomposite membrane or local insulin injection exhibit a significant increase. Conclusively, CS/ZnO nanocomposite membrane and local insulin accelerate diabetic wound healing through activation of growth factors production and stimulating the proliferation of inflammatory cells during the healing process and angiogenesis, followed by wound maturation.