Erxu Tao, Yiwei Sun, Yaxin Liu, Cailiang Shen, Yuanyin Wang, Xianwen Wang
ABSTRACT Implant‐related infections mediated by bacterial biofilm colonization remain a predominant contributor to orthopedic prosthesis failure. While conventional high‐dose antibiotic regimens frequently demonstrate limited clinical efficacy, often necessitating prosthesis replacement postdebridement, such interventions impose substantial psychological and financial burdens on patients. To address this challenge, ES@Cu 2 O nanozymes with dual‐enzyme activities were engineered by integrating cuprous oxide nanoparticles (Cu 2 O NPs) with the copper ionophore elesclomol (ES), aiming to combat implant‐related infections through enhanced cuproptosis‐like death. ES@Cu 2 O nanozymes exploit peroxidase‐like (POD‐like) activity to catalyze the conversion of endogenous hydrogen peroxide (H 2 O 2 ) into reactive oxygen species (ROS), generating a potent oxidative surge within the biofilm microenvironment. Concurrently, the glutathione‐peroxidase‐like (GSH‐Px‐like) activity of ES@Cu 2 O effectively depletes overexpressed glutathione (GSH), thereby increasing the ROS‐mediated therapeutic efficacy. Notably, ES exacerbates aberrant intracellular Cu 2 + accumulation, intensifying cuproptosis‐like death. Compared with the Cu 2 O treatment group, the ES@Cu 2 O treatment group demonstrated significantly greater antibacterial and biofilm eradication capabilities in vitro. RNA sequencing (RNA‐seq) revealed that in the ES@Cu 2 O treatment group, key energy metabolism pathways, including the TCA cycle, pyruvate metabolism, and oxidative phosphorylation, were substantially suppressed relative to those in the Cu 2 O treatment group, supporting the mechanism that ES‐mediated Cu 2 + overload potently enhances cuproptosis‐like death. In vivo, ES@Cu 2 O exhibited excellent antibacterial and restorative effects in mouse implant‐related infection models, while biosafety assessments confirmed its negligible systemic toxicity. Overall, this study revealed that the ES@Cu 2 O nanozymes with dual‐enzyme activities robustly eliminate biofilms and eradicate colonized bacteria in implant‐related infections by enhancing cuproptosis‐like death, suggesting a novel therapeutic strategy with significant potential for clinical translation.