Abstract Bacterial infections and antibiotic resistance represent significant global public health challenges, necessitating the development of innovative antibacterial agents with targeted delivery capabilities. Our study utilized macrophages’ natural ability to recognize bacteria and the increased reactive oxygen species (ROS) at infection sites to develop a novel nanoparticle for targeted delivery and controlled release. We prepared bacteria-activated macrophage membranes triggered by Staphylococcus aureus (Sa-MMs), which showed significantly higher expression of Toll-like receptors (TLRs), compared to normal macrophage membranes (MMs). These Sa-MMs were then used to coat vancomycin-loaded amphiphilic nanoparticles with ROS responsiveness (Van-NPs), resulting in the novel targeted delivery system Sa-MM@Van-NPs. Studies both In vitro and in vivo demonstrated that biocompatible Sa-MM@Van-NPs efficiently targeted infected sites and released vancomycin to eliminate bacteria, facilitating faster wound healing. By combining targeted delivery to infected sites and ROS-responsive antibiotic release, this approach might represent a robust strategy for precise infection eradication and enhanced wound healing.