The rising dangers of bacterial infections have created an urgent need for the development of a new generation of antibacterial nanoagents and therapeutics. A new near‐infrared 808 nm laser‐mediated nitric oxide (NO)‐releasing nanovehicle (MoS2‐BNN6) is reported through the simple assembly of α‐cyclodextrin‐modified MoS2 nanosheets with a heat‐sensitive NO donor N,N′‐di‐sec‐butyl‐N,N′‐dinitroso‐1,4‐phenylenediamine (BNN6) for the rapid and effective treatment of three typical Gram‐negative and Gram‐positive bacteria (ampicillin‐resistant Escherichia coli, heat‐resistant Escherichia faecalis, and pathogen Staphylococcus aureus). This MoS2‐BNN6 nanovehicle has good biocompatibility and can be captured by bacteria to increase opportunities of NO diffusion to the bacterial surface. Once stimulated by 808 nm laser irradiation, the MoS2‐BNN6 nanovehicle not only exhibits photothermal therapy (PTT) efficacy but also can precisely control NO release, generating oxidative/nitrosative stress. The temperature‐enhanced catalytic function of MoS2 induced by 808 nm laser irradiation simultaneously accelerates the oxidation of glutathione. This acceleration disrupts the balance of antioxidants, ultimately resulting in significant DNA damage to the bacteria. Within 10 min, the MoS2‐BNN6 with enhanced PTT/NO synergetic antibacterial function achieves >97.2% inactivation of bacteria. The safe synergetic therapy strategy can also effectively repair wounds through the formation of collagen fibers and elimination of inflammation during tissue reconstruction.

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