Anticandidal Activity of Hydroalcoholic Extract of Phyllanthus niruri L. (Stone-Breaker)

HIGHLIGHTS

• Hydroalcoholic extract of Phyllanthus niruri (HE-Pn) demonstrates anti-Candida activity.

• HE-Pn reduces the germ-tube formation and adhesion of Candida albicans.

• HE-Pn causes cell wall detachment, cytoplasmic granulation, vacuolation and chromatin condensation.

• HE-Pn shows no toxicity on human keratinocyte cell line.

Abstract

Candida is becoming more resistant to conventional treatments, and causes persistent and severe infections. This study evaluates the antifungal and virulence activities of the hydroalcoholic extract of Phyllanthus niruri (HE-Pn) on Candida. HE-Pn was prepared by maceration technique. Chemical composition of HE-Pn was determined using Gas Chromatography-Mass Spectrometry (GS-MS). Antifungal screening was done using agar well diffusion. CLSI M27-A3 was used to determine the Minimum Inhibitory (MIC) and Fungicidal Concentrations (MFC). Effects of HE-Pn on adhesion and germ tube of C. albicans (ATCC MYA-2876) were determined using XTT assay and germ tube formation assay, respectively. Transmission Electron Microscopy (TEM) was performed to visualize the post-exposure cellular changes. HE-Pn cytotoxicity was determined using human keratinocyte cell line (HaCaT). Chlorhexidine digluconate (2 mg/mL) was used as the positive control. Linolenic acid ethyl ester was the most abundant chemical component of HE-Pn. All strains tested were sensitive to HE-Pn. MIC were 0.03 - 8 mg/mL and MFC were 0.5 - 64 mg/mL for all test strains. C. albicans (ATCC MYA-2876) showed 50% of adhesion reduction with > 4 mg/mL of HE-Pn and germ-tube formation was inhibited with 0.25 and 2 mg/mL. TEM exhibited cytoplasmic granulation, intracellular vacuoles, detachment of cell wall and plasma membrane and chromatin condensation of Candida. No toxicity of HE-Pn was noted on HaCaT cells. HE-Pn shows an anti-Candida activity and can be used as an inhibitory agent against adhesion and germ tube formation of Candida albicans (ATCC MYA-2876) without causing any toxicity to human cells.

Keywords:
Phyllanthus niruri; hydroalcoholic; plant extract; Candida albicans; HaCaT cells; antifungal agent