Otitis media (OM) is the primary reason for pediatric antibiotic prescriptions. Nontypeable Haemophilus influenzae (NTHi) and Streptococcus pneumoniae (S. pneumoniae) are two of the most common pathogens causing OM in the middle ear, responsible for approximately 35% and 23% of all OM cases, respectively. Current treatment requires a rigorous regimen of multidose antibiotics over 5–10 days. Systemic antibiotic exposure and often prematurely terminated treatment due to the challenge of drug administration to young patients are believed to contribute to antibiotic resistance. Moreover, the development of new antibiotics has stagnated in the last ten years. In response to this crisis, nanotechnology has emerged as a promising solution to tackle drug resistance and enhance treatment outcomes. A wide range of biomimetic nanomaterials, termed nanozymes, have demonstrated potent antimicrobial efficacy. Lipid-based nanomaterials serving as delivery vehicles for antimicrobials, as well as metal-, metal oxide-, and non-metal-based nanozymes, have been utilized as therapeutics to treat bacterial infections. To address the challenges of antibiotic resistance and the lack of antibiotic development, this thesis selected metal-oxide nanoparticles with enzyme-like activity to treat OM. This class of nanozymes was chosen because their antimicrobial effects leverage the pathogens' metabolic products. The thesis began by developing a hydrogel formulation containing biocompatible silver nanoparticles with peroxidase activities, which effectively eradicated S. pneumoniae and NTHi. The formulation demonstrated minimum inhibitory concentrations (MICs) lower than or comparable to conventional antibiotics with undetectable cytotoxicity. Subsequently, the thesis developed smart therapeutics targeting OM pathogens. Vanadium pentoxide nanowires (V2O5 NWs) with haloperoxidase activities were designed to convert a metabolic product of S. pneumoniae (i.e., H2O2) into a potent antiseptic (HOBr); HOBr is only produced during an active episode of S. pneumoniae infection. A single dose of the nanowire formulation eradicated OM in chinchillas without harming tissues or affecting hearing sensitivity. Building upon that success, a cascade nanozyme was designed to target bacterial biofilms in the middle ear, which cause chronic or recurrent OM affecting one-third of U.S. children. In addition to the haloperoxidase mentioned above, glucose oxidase activity was introduced into the system by incorporating gold nanoparticles. The cascade activities converted non-ROS, like glucose and O2, into HOBr, which proved critical for mitigating bacterial biofilms in the middle ear. Considering that both pathogens coexist, the V2O5 NWs formulation was used to convert S. pneumoniae's H2O2 into a powerful antiseptic (HOBr), thereby eradicating both S. pneumoniae and NTHi simultaneously. Furthermore, the V2O5 NWs inhibited coexisting biofilms and exhibited a synergistic effect when combined with antibiotics. In conclusion, this thesis systematically investigates nanozyme-based therapeutics for the antibiotic-free treatment of OM. These therapeutics offer precise, autonomous antimicrobial synthesis and targeted drug delivery, thereby reducing antibiotic reliance and improving therapeutic outcomes for pediatric patients. The extension of this thesis will focus on achieving non-invasive drug delivery strategies. Preliminary data of a hydrogel formulation containing DSPG liposomes with encapsulated commercial antibiotics demonstrated viability for future study. This approach points to sustained drug release, enabling targeted delivery through the tympanic membrane while circumventing systemic antibiotic exposure.