Diabetic ulcers (DUs) are a chronic, non-healing diabetes complication that leads to high hospital expenses and, in extreme cases, to amputation. Peripheral vascular diseases, diabetic neuropathy, abnormal cellular and cytokine activity are among the major factors that hinder diabetic wound healing. DUs represent an important challenge in the development of new and efficient wound dressings. The extracellular matrix (ECM) has been effectively used as a scaffold for constructive remodeling of multiple tissues in animal and human. Sheep’s urinary bladder matrix was evaluated for its wound healing activity in streptozotocin-induced diabetic rats using excision model. In this experiment, 48 male Sprague dawley rats weighing 220–250g were divided into four equal groups of control, vaseline, diabetics + (10mg/wound) and diabetics+ECM2 (50mg/wound). Diabetes was induced by intraperitoneal injection of streptozotocin (45mg/kg B.W) solved in 0.05M citrate buffer. Seven days after confirming diabetes statue, skin wounds were created on the back of each rat. Rate of wound healing and histological assay using hematoxylin and Eosin staining (H&E) were used for evaluation of the wound healing in different groups. ECM treated animals exhibited significant improvement in both wound area and rate of wound healing when compared to controls (p<0.001). The ECM treated wounds were found to epithelize faster as compared to controls. The sheep’s ECM promotes significant wound healing in male diabetic rats and further studies on this activity in animal models and humans are suggested.