Anticancer Capacities of Green‐Synthesized Zinc Oxide Nanoparticles Loaded With Mangiferin: Mechanistic Insights Into Membrane Damage, pH Change, Oxidative Stress, Apoptosis, and Nuclear Lysis in HepG2 and Caco‐2 Cells

ABSTRACT Given the adverse effects of conventional chemotherapeutic agents, natural compounds and biocompatible materials have garnered growing interest in cancer therapy. Our previous research demonstrated that zinc oxide nanoparticles (ZnO NPs), synthesized through an ultrasound‐mediated green method using coffee leaves, functioned as effective nanocarriers for the controlled release of mangiferin (MGF). Here, we investigated the anticancer activity of MGF, ZnO NPs, and MGF‐loaded ZnO NPs (MGF‐ZnO NPs) on HepG2 and Caco‐2 cells, along with their underlying anticancer mechanisms. We found that both ZnO NPs and MGF‐ZnO NPs significantly reduced cell viability and increased the extracellular to intracellular pH ratio (pHe/pHi) and reactive oxygen species levels in HepG2 and Caco‐2 cells. MGF‐ZnO NPs further induced notable morphological changes and enhanced cell membrane permeability, chromatin condensation, and nuclear lysis. Additionally, both ZnO NPs and MGF‐ZnO NPs increased the overall apoptotic rate in cancer cells, with MGF enhancing the cytotoxic effects of MGF‐ZnO NPs. Molecular docking analysis revealed that MGF‐ZnO NPs may disrupt cell membrane biogenesis by inhibiting the biological activity of choline/ethanolamine phosphotransferase 1 (CEPT1), an enzyme involved in phospholipid synthesis, through hydrogen bonding and hydrophobic interactions. Additionally, MGF‐ZnO NPs also exhibited strong binding affinity to DNA bases through hydrogen bonds (DC15(B) and DG12(A)) and hydrophobic interactions (DC23(B), DG22(B), DA18(B), DA17(B), and others), potentially compromising DNA function. This study suggests that loading ZnO NPs with MGF is a promising approach to enhance cancer therapy and underscores the complex mechanisms involved in nanotherapeutic action.