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Guoqing Wan,1,2 Lingao Ruan,2,3 Yu Yin,2,3 Tian Yang,2,3 Mei Ge,2 Xiaodong Cheng1,4 1School of Life Science and Technology, China Pharmaceutical University, Nanjing, 2Shanghai Laiyi Center for Biopharmaceutical R&D, 3School of Pharmacy, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 4Department of Integrative Biology & Pharmacology, The University of Texas Health Science Center, Houston, TX, USA Abstract: Acinetobacter baumannii resistance to carbapenem antibiotics is a serious clinical challenge. As a newly developed technology, silver nanoparticles (AgNPs) show some excellent characteristics compared to older treatments, and are a candidate for combating A. baumannii infection. However, its mechanism of action remains unclear. In this study, we combined AgNPs with antibiotics to treat carbapenem-resistant A. baumannii (aba1604). Our results showed that single AgNPs completely inhibited A. baumannii growth at 2.5 µg/mL. AgNP treatment also showed synergistic effects with the antibiotics polymixin B and rifampicin, and an additive effect with tigecyline. In vivo, we found that AgNPs–antibiotic combinations led to better survival ratios in A. baumannii-infected mouse peritonitis models than that by single drug treatment. Finally, we employed different antisense RNA-targeted Escherichia coli strains to elucidate the synergistic mechanism involved in bacterial responses to AgNPs and antibiotics. Keywords: Acinetobacter baumannii, AgNPs, synergistic, antibiotic combination, anti­sense RNA