ABSTRACT: Fabrication of silver nanoparticles (AgNPs) using Chinese herbal medicine is popular as the bioactive components included in them would generate potential synergistic effect with the metal nanoparticles. The leaf of Mentha pulegium, whose extract contains a range of phytochemicals and exhibits a wide spectrum of bioactivities, is used as Chinese herbal medicine after drying naturally. Thus, the green synthesis of AgNPs using Mentha pulegium has aroused interests from analysts. However, the biosynthesis of AgNPs under alkaline conditions and the biological activities remain elusive, where alkaline conditions may influence the physicochemical properties and the biological activities of biosynthesized AgNPs. In this study, we were stimulated to fabricate bioactive AgNPs using Mentha pulegium extract under alkaline conditions, accompanied by a systematic evaluation on the effect of biosynthesis parameters on the formation, average size, and polydispersity of AgNPs. Our results showed that alkaline conditions could accelerate the formation of AgNPs with a small average size but at a disadvantage to the polydispersity. Additionally, the as-prepared AgNPs had a hexagonal structure and spherical shape with an average size of 15.7 ± 0.1 nm, existing in the monodispersed form and revealing a high degree of stability. The AgNPs exhibited potent antioxidant and significant inhibitory activity for both bacterial and cancer cell lines. The MIC values of AgNPs for Staphylococcus aureus and Escherichia coli were both 50.0 μg·mL-1, and the IC50 values for HCT116, HepG2, and HeLa cells were 9.0, 14.5, and 31.5 μg·mL-1, respectively. The AgNPs biosynthesized using M. pulegium under alkaline conditions, which had a smaller size and more surface loads, are entirely different with those synthesized under acidic conditions, and the anticancer activity increased significantly. The internalization of AgNPs inside these five cells displayed a variant trend with variable AgNPs concentrations, suggesting the different mechanism of cell death. For two pathogens, HCT116 and HepG2 cancer cell lines, both cell wall and intracellular damage may be responsible for the cell death. However, for Hela cell line the cell death may be rooted in oxidative stress or intracellular penetration. These results confirmed that the AgNPs biosynthesized from M. pulegium extract under alkaline conditions would act as better anticancer agents in biomedicine.
