Biofilms are complex bacterial communities that significantly hinder the treatment of chronic and recurrent infections by enhancing bacterial virulence and conferring resistance to antimicrobial therapies. To address this challenge, an intelligent DNA nanomedicine has been engineered to dismantle biofilms and target resistant bacteria, offering an innovative solution for chronic wound infections. These nanomedicines initiate biofilm degradation through in situ generation of potent oxidative radicals, enabling deep biofilm penetration and precise bacterial targeting. Utilizing aptamers for specific bacterial identification, the nanomedicines concentrate therapeutic agents directly at infection sites. The combined effect of severe oxidative stress and sustained silver ion release ensures a continuous, focused assault on pathogens, effectively eradicating resistant bacteria. This strategy demonstrated broad-spectrum efficacy against both Gram-positive and Gram-negative bacteria, significantly enhancing wound healing in a diabetic infection model. By integrating intelligent bacterial eradication with modulation of the wound microenvironment, this approach presents a promising solution for overcoming biofilm-associated resistance and advancing chronic wound infection treatment.