The widespread increase in antibiotic resistance (AR), in an extensive range of microorganisms, demands the development of alternative antimicrobials with novel non-specific low-mutation bacterial targets. Silver nanoparticles (AgNPs) and photosensitizers (PSs) are promising antimicrobial agents with broad-spectrum activity and low tendency for antimicrobial resistance development. Herein, we investigated the light-mediated oxidation of AgNPs for accelerated release of Ag⁺ in the antibacterial synergy of PS-AgNP conjugates using protoporphyrin IX (PpIX) as a PS. Also, the influence of polyethyleneimine (PEI) coated AgNPs in promoting antibacterial activity was examined. We synthesized, characterized and tested the antimicrobial effect of three nanoparticles: AgNPs, PpIX-AgNPs, and PEI-PpIX-AgNPs against a methicillin-resistant Staphylococcus aureus strain (MRSA) and a wild-type multidrug resistant (MDR) E. coli. PpIX-AgNPs were the most effective material achieving >7 log inactivation of MRSA and MDR E. coli. The order of bacterial log inactivation was PpIX-AgNPs > PEI-PpIX-AgNPs > AgNPs. This order correlates with the trend of Ag⁺ concentration released by the NPs (PpIX-AgNPs > PEI-PpIX-AgNPs > AgNPs). Our study confirms a synergistic effect between PpIX and AgNPs in the inactivation of AR pathogens with about 10-fold increase in inactivation of ARB relative to AgNPs only. The concentration of Ag⁺ released from NPs determined the log inactivation of MRSA and MDR E. coli more than either the phototoxic effect or the electrostatic interaction promoted by surface charge of nanoparticles with bacteria cells. All NPs showed negligible cytotoxicity to mammalian cells at the bacterial inhibitory concentration after 24 h exposure. These observations confirm the crucial role of optimized Ag⁺ release for enhanced performance of AgNP-based antimicrobials against AR pathogens.

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