Current antibiotic treatments fail to eliminate the
Clostridium difficile
(
C. difficile
) spores and induce dysbiosis and intestinal inflammation via off-target effect, which causes refractory
C. difficile
infection raise an unmet need for a spore-specific antimicrobial treatment. We developed a sporicidal and antimicrobial vancomycin-loaded spore-targeting iron oxide nanoparticle (van-IONP) that selectively binds to
C. difficile
spores. Cryo-electron microscopy showed that vancomycin-loaded nanoparticles can target and completely cover spore surfaces. They not only successfully delayed the germination of the spores but also inhibited ∼50% of vegetative cell outgrowth after 48 h of incubation. The van-IONPs also inhibited the interaction of spores with HT-29 intestinal mucosal cells
in vitro
. In a murine model of C. difficile infection, the van-IONP significantly protected the mice from infected by
C. difficile
infection, reducing intestinal inflammation, and facilitated superior mucosal viability compared with equal doses of free vancomycin. This dual-function targeted delivery therapy showed advantages over traditional therapeutics in treating
C. difficile
infection.
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