Objective:To evaluate the graft success rate and postoperative hearing gain for tympanic membrane perforation? using Human-derived acellular dermal matrix under otoendoscope. Methods:A total of 28 patients who had received endoscopic myringoplasty without tympanomeatal flap elevation in our department from November? 2023 to January? 2024 were included. The recovery of tympanic membrane and hearing after surgery were examined using “before-after study in the same patient” design. Preoperative high-resolution computed tomography (HRCT) of temporal bone, endoscopic ear examination, pure tone audiometry and other routine examinations were performed. The outcomes included graft success evaluated by endoscopy and hearing gain evaluated by pure tone audiometry at 0.5,1 and 3 months after the surgery. The therapeutic efficiency was analyzed using SPSS software 27. Results: Three months after the surgery, the graft success rate achieved 93% (26/28). The preoperative air conduction hearing 41.61± 9.40dB HL was significantly greater than postoperatively 28.75±5.68dB HL at 3 months (P=0.009). Conclusion: Otoendoscopic repair of tympanic membrane perforation with Human-derived acellular dermal matrix as an implant material is a reliable treatment with few complications and good postoperative hearing recovery.