Synergistic effects of tea polyphenol epigallocatechin 3-O-gallate and azole drugs against oral Candida isolates

Abstract

Background: The antifungal drug resistance has become an emerging problem in the management of candida infections worldwide. The objective of this study was to examine the efficacy of epigallocatechin 3-O-gallate (EGCG) alone and in combination with fluconazole/ketoconazole drugs against oral Candida isolates.

Methods: Minimum inhibitory concentration (MIC) and minimum fungicidal concentrations (MFC) of EGCG against 60 oral Candida isolates and 4 ATCC strains were determined. Synergism of EGCG with azole drugs was evaluated by checkerboard micro-dilution method and calculated fractional inhibitory concentration index (FICI). Candida cells' ultrastructure was studied by electron microscopy.

Results: MIC and MFC values of EGCG were in the range of 3.91-15.63 and 15.63-31.25μg/mL, respectively. Minimum biofilm inhibitory concentration (MBIC) range of EGCG (62.5-125μg/mL), was less than the ketoconazole (64-256μg/mL) and fluconazole (128-512μg/mL). The combination of EGCG with fluconazole/ketoconazole exhibited synergistic effects (ΣFICI≤0.50). EGCG with azole drugs showed high sensitivity against the tested isolates in growth curve assays. Against the biofilm, the susceptibility of fluconazole/ketoconazole significantly increased (3 to 5 fold), after combination with EGCG (MBIC/4) (P≤0.001). Electron microscopy of EGCG treated cells showed deformation of cell structure, ruptured cell wall and release of intracellular content. In molecular docking experiments, a strong interaction was observed between EGCG and fungal cell membrane molecule ergosterol.

Conclusion: We conclude that EGCG synergistically enhanced the antifungal potential of azole drugs. The synergistic potential of EGCG might be helpful in preventing the development of drug resistance, in lowering the drug dosage, and thus minimizing adverse effects.

Keywords: Antifungal; Epigallocatechin 3-O-gallate; Microscopic analysis; Oral candidiasis; Synergism.