Objective To investigate the efficacy of endoscopic cartilage packing in the treatment of cholesteatoma in the outer and middle ear. Methods Twenty-six patients (26 ears) with cholesteatoma who underwent surgery from January 2019 to January 2023 were collected. The epitympanic cholesteatoma was completely removed, and the epitympanic was filled with cartilage (tragus, concha cavity or nasal septum cartilage). The endoscopic examination, pure tone audiometry and acoustic immittance were reexamined at 6 months after operation. One year after operation, otoendoscopy was performed. Results The preoperative air conduction pure tone audiometry was (41.25±2.39dBHL), and it was (26.32±4.34 dBHL) 6 months after operation. Air-bone gap (ABG) was (26.45±5.39 dBHL) before operation and (12.69±6.43 dBHL) 6 months after operation. Six months after operation, there was no invagination of the lateral wall of the attic and no recurrence of cholesteatoma. One year after operation, 1 case had invagination, and cholesteatoma recurred, 1/26 (3.84%). Conclusion Cartilage packing can effectively prevent the lateral epitympanic retraction, reduce the recurrence of cholesteatoma, and improve the hearing after operation.