Synergistic effects of Thymus vulgaris essential oil in combination with antifungal agents and inhibition of virulence factors of Candida albicans

BLANC, ALAN ROY; SORTINO, MAXIMILIANO ANDRÉS; BUTASSI, ESTEFANÍA; SVETAZ, LAURA ANDREA

Phytomedicine Plus

Elsevier

Lugar: Amsterdam; Año: 2023

2667-0313

Background: Combination antifungal therapy has become a prominent approach in medical practice as it takesadvantage of synergistic interactions by interfering with multiple targets of the pathogen, broadening thespectrum of activity, reducing the development of resistance, and improving treatment outcomes. Therefore,combining conventional antifungal drugs with natural products can increase antifungal activity, reduce sideeffects, and optimize therapeutic effects.Purpose: The present study aimed to evaluate the antifungal activity of thyme essential oil (ThyEO) in combination with amphotericin B, caspofungin, fluconazole, itraconazole, and posaconazole (PSZ), as well as itsinhibitory effect on Candida albicans virulence factors.Materials and Methods: The microbroth dilution assay was employed to determine the minimum inhibitoryconcentration (MIC) and minimum fungicidal concentration (MFC). Interactions were assessed using themicrodilution checkerboard assay. Ergosterol and methylene blue assays were used to detect effects on fungalmembrane, while the sorbitol assay was used to detect effects on fungal cell wall. Inhibition of yeast virulencefactors (adherence to epithelial cells, germ tube and pseudomycelium formation, secretion of hydrolytic enzymes, and biofilm formation) was assessed with previously reported methods.Results: Almost all combinations showed additivism against C. albicans, except ThyEO/PSZ (31.25/0.0039 µg/ml), which showed partial synergism. Furthermore, all mixtures were fungicidal against C. albicans strains.ThyEO/PSZ, its components alone, and thymol were shown to disrupt the fungal cytoplasmic membrane,increasing its permeability. ThyEO/PSZ, at sub-inhibitory concentrations, significantly decreased the ability ofC. albicans to adhere to buccal epithelial cells. ThyEO/PSZ, ThyEO, and PSZ were able to reduce the pseudomycelium production of C. albicans while thymol completely inhibited its formation. ThyEO/PSZ, each combination component on its own, and thymol inhibited biofilm formation and preformed biofilm of C. albicans.Notably, ThyEO/PSZ showed synergistic and fungicidal activity against a resistant strain of C. albicans, reducingthe PSZ dose by 4-fold.Conclusion: These findings make ThyEO and ThyEO/PSZ mixture valuable candidates for the development ofalternative antifungals with a lower incidence of adverse effects