Quatsome nanovesicles as antibacterial platform: Mechanistic insights into their activity against planktonic and biofilm Staphylococcus aureus.

The growing threat of antibiotic-resistant pathogens has intensified the demand for alternative antibacterial materials. Quatsomes-nanovesicles composed of cholesterol and quaternary ammonium surfactants (QAS)-emerge as promising candidates due to their intrinsic antimicrobial properties and tunable physicochemical characteristics. Here, we investigate the antibacterial activity of quatsomes incorporating QAS with either tetradecyl (C14) or hexadecyl (C16) alkyl chains against Staphylococcus aureus, a leading cause of hospital-acquired infections. Both quatsome types exhibited potent bactericidal activity in planktonic cultures, with C16-containing formulations showing a 2.5-fold lower minimum bactericidal concentration than C14 counterparts. Confocal microscopy suggested a partial penetration of cationic quatsomes into the bacterial peptidoglycan layer, accompanied by significant increases in ζ-potential, suggesting strong electrostatic interactions without visible membrane disruption, as confirmed by scanning electron microscopy. Both formulations also demonstrated high efficacy against mature S. aureus biofilms, with no significant differences between alkyl chain lengths, indicating a mechanism primarily targeting the extracellular biofilm matrix. In addition, they showed a good antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA). A preliminary safety assessment using reconstructed human epidermis (Episkin™) confirmed the non-irritant nature of both formulations. These findings highlight the potential of QAS-based quatsomes as effective and biocompatible nanocarriers for topical antibacterial applications, offering a promising platform for combating antibiotic-resistant infections in both planktonic and biofilm states.