BACKGROUND: Periprosthetic joint infection (PJI) is a significant complication following arthroplasty, attributed to the biofilm formation. This study evaluates the effectiveness of vancomycin-loaded microbubbles (Van-MBs) in conjunction with ultrasound-targeted microbubble destruction (UTMD) on biofilm disruption and bactericidal efficiency. METHODS: Van-MBs were prepared using the thin-film hydration method and characterized using microscopy, dynamic light scattering analysis, and high-performance liquid chromatography (HPLC). Confocal laser scanning microscopy (CLSM) was used to assess the penetration of Van and Van-MBs into biofilms. Biofilms were treated with Van, Van-MBs, UTMD, and Van-MBs + UTMD. CLSM and crystal violet staining were utilized to assess the morphology, viability, and biomass of the biofilms. Bacterial activity was examined through scanning electron microscopy (SEM) and plate counting, while gene expression was analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: The results demonstrated that Van-MBs penetrated deeper into methicillin-resistant CONCLUSION: These findings demonstrate the potential of Van-MBs in combination with UTMD as an innovative approach to enhance antibiotic efficacy and eliminate biofilms in the treatment of PJI.