Photocatalytic Activation Strategy for Sequential Prodrug Release to Overcome Resistant Biofilms

Drug-resistant biofilms present a formidable therapeutic challenge by limiting the antibiotic penetration and enhancing bacterial tolerance. A rational approach is to first disperse the biofilm to sensitize the embedded bacteria, followed by efficient antibiotic-mediated eradication. Such precisely ordered sequential release is essential to synchronizing biofilm dispersion with bacterial killing, thereby maximizing therapeutic efficacy while minimizing infection spread. However, achieving temporally resolved, time-gated release of distinct therapeutic agents remains highly challenging, particularly within a single-component system. Here, we report a nanoconfined photocatalytic platform that enables the sequential release of nitric oxide (NO) and levofloxacin (LEV) from a single-component prodrug. This strategy ensures an initial NO-mediated downregulation of antibiotic-resistant gene expression and enhanced bacterial susceptibility, followed by LEV-mediated bacterial eradication. The synchronized therapeutic sequence achieved potent antibiofilm efficacy in diabetic mice infected with LEV-resistant Pseudomonas aeruginosa, resulting in biofilm eradication, accelerated wound closure, and attenuated inflammation. Beyond antibacterial therapy, this one-component sequential release paradigm establishes a generalizable strategy for temporally programmed delivery of chemotherapeutic, anti-inflammatory, or anesthetic agents, underscoring both its conceptual novelty and broad translational potential.