A significant percentage of tympanic membrane (TM) perforations require some form of therapy to heal. Topical application of epidermal growth factor (EGF), a potent mitogen for epidermal and mesodermal cells, promotes healing of injuries in tissues histologically similar to TM, such as cornea and skin. We evaluated TM as a target tissue for EGF action. Specific, high affinity receptors for EGF were detected in TM (Kd = 3.1 nM, 150 fmol receptor/mg protein). Autoradiography of iodine 125-EGF binding to intact TMs revealed that EGF receptors were present on cells in the stratified squamous epithelial layer and in the stromal/mucoepithelial layer. Repetitive treatment of perforated cat TMs with 20 μg of EGF formulated in saline induced substantial hyperplasia of epithelial and stromal layers compared to paired TMs treated with saline. A single treatment of perforated cat TMs with 50 μg of EGF formulated in a hydrogel or in shredded Gelfoam produced significantly (p< .05) smaller perforations at 6 days following the operation compared to paired TMs treated with vehicles. These results demonstrate that TM is a target tissue for EGF and that topical treatment with EGF stimulates healing of TM perforations.