The cascade catalytic strategy could effectively enhance the antibacterial activity by regulating the production of hydroxyl radicals (˙OH) in the sites of bacterial infection. In this work, a ruthenium metal nanoframe (Ru NF) was successfully synthesized via the palladium template method. The cascade catalysis in the bacterial infection microenvironment was achieved by physically adsorbed natural glucose oxidase (GOx), and hyaluronic acid (HA) was coated on the outer layer of the system for locating the infection sites accurately. Eventually, a composite nano-catalyst (HA-Ru NFs/GOx) based on the ruthenium nanoframe was constructed, which exhibited excellent cascade catalytic activity and good biocompatibility. The prepared HA-Ru NFs/GOx enhances the antibacterial activity and inhibits bacterial regeneration through the outbreak of reactive oxygen species (ROS) caused by self-activating cascade reactions. In addition, in vivo experiments indicate that HA-Ru NFs/GOx could efficiently cause bacterial death and significantly promote wound healing/skin regeneration. Accordingly, ruthenium metal framework nanozymes could be used as an effective cascade catalytic platform to inhibit bacterial regeneration and promote wound healing, and have great potential as new antibacterial agents against antibiotic-resistant bacteria.