Programmable Engineered Living Vehicles as Autonomous Biofactories for Synergistic Osteomyelitis Management and Bone...

Conventional static biomaterials and synthetic nanomedicines struggle to penetrate recalcitrant biofilms and orchestrate tissue regeneration in deep-seated infections due to limited autonomous navigation and a lack of spatiotemporal responsiveness. Here, we conceptualize and develop a programmable engineered living vehicle (ELV) that functions as an autonomous biofactory for the synergistic management of drug-resistant osteomyelitis. Built upon a self-propelled Escherichia coli Nissle 1917 chassis with innate ecological dominance, these living biofactories are hierarchically equipped with environmental biosensors and logic-gated computing modules. Crucially, leveraging AlphaFold 3-guided molecular docking, we engineered multiscale thermo-responsive genetic logic circuits that enable precise, temperature-dependent biomanufacturing of bacteriolytic lysostaphin (LYS) and vasoactive intestinal peptide (VIP) (≥37°C), coupled with a fail-safe thermal self-destruct mechanism (≥42°C). Furthermore, surface-engineered (DSS)₆ peptides confer robust hydroxyapatite affinity, acting as a bone-homing navigation module. In a rat osteomyelitis model induced by methicillin-resistant Staphylococcus aureus (MRSA), intravenously deployed ELV biofactories autonomously navigate to the infected niche, actively penetrate biofilms, and outperform vancomycin in pathogen clearance. Concurrently, localized payload secretion reprograms the osteo-immune microenvironment, significantly accelerating bone repair. This work establishes a versatile paradigm for next-generation smart living therapeutics, bridging synthetic biology with micro-engineering to address complex clinical challenges.