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

ABSTRACT 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.