Development and Characterization of Genistein-Loaded Silver Nanoparticles for the Induction of Apoptosis on Human Hepatocellular Carcinoma Cell Lines

The defect of the apoptotic process is one of the most important causes that contribute to hepatocarcinoma (HCC) concerning cancer progression. Chemotherapeutic approaches are designed to modulate apoptosis; however, its toxic side effect needs to develop alternative supportive therapy. In this concern, targeted phytoconstituent-loaded nanocarriers are highlighted as an alternative supportive therapy with fewer side effects and regulates apoptosis. The study comprised the development of the genistein-loaded silver nanoparticles (G-AgNPs) and determined the in vitro apoptosis effect. The chemical reduction method was employed for the preparation of G-AgNPs and optimized by using the Box-Behnken design (BBD). The F3 batch shows a zeta potential value of - 29.37 +/- 1.83 mV, with particle size 123.47 +/- 3.76 nm, and % entrapment efficiency of 96.82 +/- 1.05% and hence considered as an optimized one. It was also characterized for solid-state physicochemical characterization and performed in vitro release of genistein at pH 6.8. The result found that G-AgNPs showed 98.05% release in 24 h by following the Higuchi release mechanism. The G-AgNPs were further evaluated to determine the in vitro IC50 (25 mu g/ml), % apoptosis (56.38%), and drug uptake (13.56 +/- 0.956) by using a human hepatocellular (HepG2) cell line. The G-AgNPs observed enhanced % apoptosis in HepG2 compared to standard camptothecin and pure genistein (42.67%). The confocal microscopical analysis and flow cytometry also revealed its significant internalization of genistein in HepG2 cells, accompanied by notable signs of apoptosis. Therefore, G-AgNPs play a substantial role in hepatocellular carcinoma by promoting apoptosis and therapeutic efficacy of genistein.