For the effective treatment of chronic otitis media caused by methicillin resistant Staphylococcus aureus (MRSA), the local delivery of vancomycin was attempted based on a formulation with pluronic F-127. The phase transition property of pluronic F-127, which is liquid at room temperature and becomes solid at body temperature, enabled convenient injection and reduced the side effects of the potent antibiotic. The temperature responsive polymer–vancomycin matrix system was also shown to effectively deliver vancomycin locally to the infected inner ear, plus the MRSA growth was inhibited by the formulation. The otitis media was completely cured by the sustained local delivery of vancomycin from the matrix in vivo. Accordingly, this system has potential clinical application for treating chronic otitis media with improved patient compliance and better therapeutic efficacy.