Pseudomonas aeruginosa (P. aeruginosa), a drug-resistant Gram-negative
pathogen, is listed among the “critical”
group of pathogens by the World Health Organization urgently needing
efficacious antibiotics in the clinics. Nanomaterials especially silver
nanoparticles (AgNPs) due to the broad-spectrum antimicrobial activity
are tested in antimicrobial therapeutic applications. Pathogens rapidly
develop resistance to AgNPs; however, the health threat from antibiotic-resistant
pathogens remains challenging. Here we present a strategy to prevent
bacterial resistance to silver nanomaterials through imparting chirality
to silver nanoclusters (AgNCs). Nonchiral AgNCs with high efficacy
against P. aeruginosa causes heritable resistance,
as indicated by a 5.4-fold increase in the minimum inhibitory concentration
(MIC) after 9 repeated passages. Whole-genome sequencing identifies
a Rhs mutation related to the wall of Gram-negative
bacteria that possibly causes morphology changes in resistance compared
to susceptible P. aeruginosa. Nevertheless, AgNCs
with laevorotary chirality (l-AgNCs) induce negligible resistance
even after 40 repeated passages and maintain a superior antibacterial
efficiency at the MIC. l-AgNCs also show high cytocompatibility;
negligible cytotoxicity to mammalian cells including JB6, H460, HEK293,
and RAW264.7 is observed even at 30-fold MIC. l-AgNCs thus
are examined as an alternative to levofloxacin in vivo, healing wound
infections of P. aeruginosa efficaciously. This work
provides a potential opportunity to confront the rising threat of
antimicrobial resistance by developing chiral nanoclusters.