The tympanic membrane plays an important role in the human hearing system, which is easily perforated under unfavorable conditions, leading to loss of hearing and otitis media. Many autologous materials and artificial materials have been used to repair a perforated tympanic membrane, but these materials sometimes can cause severe hearing loss because of their adhesion to the ossicle during the healing process and the postoperative process. Herein, we report Janus membranes with asymmetric cellular adhesion behaviors for regenerating the eardrum. These Janus membranes are constructed by co-depositing a tannic acid (TA)/3-aminopropyltriethoxysilane (APTES) coating on one surface of the polypropylene microfiltration membrane. Cellular experiments indicate that the Janus membranes have good biocompatibility and asymmetric cellular adhesion properties. The repair of the tympanic membrane perforation experiment and laser Doppler vibrometer (LDV) measurements prove that the hydrophilic surface of Janus membranes repairs perforated eardrums, and meanwhile the hydrophobic surface can avoid adhering to the inner ear tissue for reducing hearing loss. The Janus membranes have good prospects in the treatment of tympanic membrane perforation.