This study looked at how the synthetic chitosan-AgNPs-Doxorubicin-folic acid combination affected the A549 cell line in terms of cytotoxicity and anticancer activity. By reducing silver nitrate (AgNO3) and biodegradable chitosan, silver nanoparticles were biosynthesized. The produced conjugate was examined by using FT-IR spectroscopy, atomic force microscopy (AFM), and field emission scanning electron microscopy (FE-SEM). The cytotoxicity assay for the viability of A549 cells revealed that the combination of chitosan, AgNPs, doxorubicin, and folic acid decrease cell viability in a dose-determined by method over 48 hours, which direct to a dependent reduce in the activity of A549 cells. The mechanism analysis of the impacted living cells leading to apoptosis revealed a considerable rise in nuclear concentration, cytochrome c, and cell membrane permeability (dose-dependent). The bright green chromatin in DOX-treated cells was compacted or broken up, indicating an early stage of apoptosis. However, cells treated with the CS-AgNPs-DOX-FA compound displayed orange nuclei and late stage apoptosis. The findings demonstrated that A549 lung cancer cells are cytotoxic to Cs-Ag NPs-DOX-FA. The Cs-Ag NPs-DOX-FA MTT assay demonstrated that the harmful effect of 25 µg/mL on A549 cells is dose-dependent, and a rise in nuclear intensity, membrane permeability, and cytochrome were observed. Cell viability also declined, and the potential of the mitochondrial membrane changed. The fact that the release of DOX was delayed shows that nanoparticles in drug carriers may be used to reduce the exposure of healthy tissues; however, boosting the accumulation to therapeutic medicine in the tumor site.
