Ozonated Oil Nanoemulsion Induces Ferroptosis and Metabolic Disruption in Candida albicans via Membrane Destabilization

The poor stability and limited water dispersibility of ozonated oil constrain its antifungal applications. To overcome these limitations, we formulated an ozonated oil nanoemulsion (Nano-Ozo) and elucidated its antifungal mechanism against Candida albicans. Nano-Ozo was characterized by particle size, zeta potential, microstructure, and ozone retention capacity. Antifungal efficacy was assessed by minimum inhibitory concentration (MIC), membrane integrity assays, reactive oxygen species (ROS) detection, metabolomics, and a murine model of vulvovaginal candidiasis (VVC). The nanoemulsion had a uniform size (98.28 ± 5.23 nm), a high zeta potential (53.20 ± 0.28 mV), and prolonged ozone stability. It showed potent antifungal activity (MIC = 31.25 μL/mL) by disrupting membrane integrity, provoking an ROS burst, and inducing ferroptosis, as indicated by changes in SOD, MDA, LDH, and GSH levels. Metabolomic analysis revealed the inhibition of glycolysis and the TCA cycle. In the VVC model, Nano-Ozo markedly reduced vaginal fungal burden and ameliorated tissue inflammation. Given its excellent stability, biological efficacy, and multi-targeted antifungal action, Nano-Ozo holds great potential for developing localized antifungal therapies, such as topical or vaginal formulations. These findings underscore Nano-Ozo as a promising nanoengineering-based antifungal strategy.