Synergy of green-synthesized silver nanoparticles and Vatica diospyroides fruit extract in inhibiting Gram-positive bacteria by inducing membrane and intracellular disruption

Abstract

Silver nanoparticles (AgNPs) are used in biomedicine applications. Other drugs combined with the AgNPs can improve efficacy in the treatment of diseases, and most such studies have focused on antibiotics. We determined the synergistic effects of Phyllanthus emblica-derived AgNPs in combination with Vatica diospyroides cotyledon extracts (VCE) against bacteria using agar well diffusion, broth microdilution, and minimum inhibitory concentration (MIC). Synergy of AgNPs and VCE was confirmed with the fractional inhibitory concentration index (FICI). To evaluate patterns of bacterial death, flow cytometry and electron microscopy were used. We found that the effective incubation time of AgNPs against bacteria was highly variable. Increasing AgNPs in the combination influenced antibacterial activity against Staphylococcus aureus and Bacillus subtilis. The MIC values interpreted through FICI showed synergy against S. aureus and indifference against B. subtilis. Flow cytometric profiles confirmed that the fraction of S. aureus that respond to a combination of VCE with AgNPs increased in dose-dependent manner. The response patterns of bacteria proceeded simultaneously as the cells lost intracellular components and suffered membrane damage. Synergy of AgNPs with a plant extract has become a promising approach, as green AgNPs and plant extracts are biocompatible and cost-effective resources that can utilized for the treatment of bacterial infectious diseases.