Eicosapentaenoic acid as an antibiofilm agent disrupts mature biofilms of Candida albicans

The biofilm formation of <i>Candida albicans</i>, a major human fungal pathogen, represents a crucial virulence factor during candidiasis. Eicosapentaenoic acid (EPA), a polyunsaturated fatty acid, has emerged as a potential antibiofilm agent against <i>C</i>. <i>albicans</i>. Herein, we aim to investigate the antifungal effect of EPA (1 mM) on the mature biofilm of <i>C. albicans</i> and explore the underlying mechanism. Crystal violet and XTT assays showed that EPA exerted a strong inhibitory efficacy on preformed biofilms in <i>C. albicans</i>. Biofilm architecture and cell viability were observed using scanning electron microscopy and confocal laser scanning microscopy, indicating that EPA could block the yeast-to-hypha transition and damage the structure, thereby exhibiting antibiofilm activity. RNA sequencing analysis revealed that EPA treatment led to the downregulation of genes associated with hyphal formation and biofilm development. From the signaling pathway perspective, EPA regulated the <i>C. albicans</i> biofilms involving two signaling pathways, namely, Ras1-cAMP-PKA and Cek-MAPK pathways. Additionally, the EPA could effectively reduce the production of key messenger cAMP in the Ras1-cAMP-PKA pathway. Interestingly, in response to EPA, ergosterol biosynthesis-related genes were down-regulated, indicating EPA as antifungal agent might reduce the risk of developing drug resistance. The findings of this study highlight the potential of EPA as an alternative or adjunctive antibiofilm agent against <i>C. albicans</i>-related infections.