An Ultrasound‐Activated Dual‐Targeted Strategy Integrating Targeted Biofilm Damage and Gluconeogenesis Inhibition for Enhanced Biofilm Eradication

ABSTRACT

Sonodynamic therapy (SDT) for bacterial infections primarily relies on high levels of reactive oxygen species (ROS) to kill bacteria. However, excessive ROS can also damage normal tissues. Herein, this study proposes a concept of an ultrasound (US)‐activated dual‐targeted antibacterial strategy that integrates targeted biofilm damage with gluconeogenesis inhibition. The US‐responsive antibacterial platform (HfO _2‐x @2‐ML), composed of defect‐engineered hafnium oxide (HfO _2‐x ), linolenic acid (LA), and a gluconeogenesis inhibitor (modified 2‐mercaptothiazole, 2‐MT), was constructed on titanium implants. This platform enables efficient biofilm eradication while potentially reducing collateral tissue damage. LA preferentially associates with bacterial membranes and synergizes with the polarization‐induced electric field to amplify membrane‐localized ROS‐driven oxidative damage, which may facilitate 2‐MT entry and inhibit FBPase‐associated gluconeogenesis. Molecular dynamics simulations indicated that LA interacted with membrane‐associated protein (6S7V) and fatty acid‐binding protein (4X9X), while transcriptomic analysis showed that FBP gene expression was downregulated after HfO _2‐x @2‐ML + US treatment. HfO _2‐x @2‐ML aggravated membrane lipid peroxidation, decreased ATP levels, and achieved a 99.78 ± 0.11% antibacterial rate against methicillin‐resistant Staphylococcus aureus (MRSA). Furthermore, Piezo1‐associated Ca ²⁺ signaling may contribute to US‐induced osteogenesis, supporting a potentially cytocompatible strategy to reduce recurrence from residual bacteria.