Antibiofilm and Membrane-Damaging Potential of Cuprous Oxide Nanoparticles against Staphylococcus aureus with Reduced Susceptibility to Vancomycin

The antimicrobial effects of copper ions and salts are well known, but the effects of cuprous oxide nanoparticles (Cu2O-NPs) on staphylococcal biofilms have not yet been clearly revealed. The present study evaluated Cu2O-NPs for their antibacterial and antibiofilm activities against heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) and vancomycin-intermediate S. aureus (VISA). Nanoscaled Cu2O, generated by solution phase technology, contained Cu2O octahedral nanoparticles. Field emission electron microscopy demonstrated particles with sizes ranging from 100 to 150 nm. Cu2O-NPs inhibited the growth of S. aureus and showed antibiofilm activity. The MICs and minimum biofilm inhibitory concentrations ranged from 625 μg/ml to 5,000 μg/ml and from 2,500 μg/ml to 10,000 μg/ml, respectively. Exposure of S. aureus to Cu2O-NPs caused leakage of the cellular constituents and increased uptake of ethidium bromide and propidium iodide. Exposure also caused a significant reduction in the overall vancomycin-BODIPY (dipyrromethene boron difluoride [4,4-difluoro-4-bora-3a,4a-diaza-s-indacene] fluorescent dye) binding and a decrease in the viable cell count in the presence of 7.5% sodium chloride. Cu2O-NP toxicity assessment by hemolysis assay showed no cytotoxicity at 625 to 10,000 μg/ml concentrations. The results suggest that Cu2O-NPs exert their action by disruption of the bacterial cell membrane and can be used as effective antistaphylococcal and antibiofilm agents in diverse medical devices.