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Biofilm infections in wounds seriously delay the healing process, and methicillin-resistant <i>Staphylococcus aureus</i> is a major cause of wound infections. In addition to inactivating micro-organisms, low-temperature gas plasma can restore the sensitivity of pathogenic microbes to antibiotics. However, the combined treatment has not been applied to infectious diseases. In this study, low-temperature gas plasma treatment promoted the effects of different antibiotics on the reduction of <i>S. aureus</i> biofilms in vitro. Low-temperature gas plasma combined with rifampicin also effectively reduced the <i>S. aureus</i> cells in biofilms in the murine wound infection model. The blood and histochemical analysis demonstrated the biosafety of the combined treatment. Our findings demonstrated that low-temperature gas plasma combined with antibiotics is a promising therapeutic strategy for wound infections.