Diabetes put off healing method as it impairs every section of wound restoration i.e. Hemostasis, irritation, proliferation and remodeling section.Insulin and a natural polymer of Chitosan (CS) can potentially repair the integrity of broken pores and skin in the field of wound repair. Zinc oxide (ZnO) possesses both antibacterial and anti-inflammatory properties and accelerates the wounds restoration. The wound recovery capability of insulin injection with chitosan/Zno nanocomposite membrane in diabetic rats were evaluated. Diabetes was induced by STZ administered intraperitoneally (i.p.) as one dose (50 mg/kg b.wt). Forty eight male rats used and divided into six groups. Group I: control wounded, non-treated , non-diabetic rats, Group II : wounded diabetic non treated, Group III : Normal Wound treated with chitosan/ Zinc oxide nanocomposite membrane, Group IV:Diabetic wounded treated with chitosan/ Zinc oxide nanocomposite membrane, Group V: wounded diabetic rats treated with local insulin injection, Group VI : Wounded diabetic treated with chitosan/ Zinc oxide nanocomposite membrane and local insulin injection. A significant decrease in (PI3K), (MAPK) genes expression in addition to miRNA 125 and miRNA 132 in skin tissue of diabetic wounds compared with normal wound. However, increase in PI3K, MAPK, miRNA 125 and miRNA 132 genes were observed in diabetic wounds treated with chitosan/Zno nanocomposite membrane or local insulin injection as compared to diabetic wounds non treated. The present data indicated that local insulin injection in wound area with chitosan /Zno nanocomposite membrane might activate the PI3K/MAPK signalling pathway and some miRNA that promote and accelerates diabetic wound healing.