Nanomaterials, encompassing nanoparticles and biopolymers, are gaining recognition as potential therapeutic agents. This study explored the antioxidant and anticancer effects of zinc oxide and selenium co-loaded chitosan/alginate bio-composites against the HeLa human cervical cancer cell line. Zinc oxide and selenium nanoparticles were synthesized using a green synthesis approach and subsequently integrated into a chitosan/alginate bio-composite, designated as hydro ZnO/Se NPs. The physicochemical characteristics were analyzed through Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), dynamic light scattering (DLS), zeta potential measurements, and electron microscopy (SEM/TEM). The antioxidant efficacy of the hydro ZnO/Se NPs was assessed via a DPPH scavenging assay. Human dermal fibroblast (HDF) and HeLa cells were subjected to various concentrations of hydro ZnO/Se NPs, with evaluations of cell viability, migration, and colony formation conducted through MTT, wound healing, and clonogenic assays, respectively. Additionally, quantitative polymer ase chain reaction (qPCR) was utilized to examine the expression of apoptosis-related genes, specifically BCL2, caspase3, caspase8, and caspase9. Characterization techniques confirmed the successful synthesis of pure, crystalline, and spherical hydro ZnO/Se NPs. Antioxidant assays revealed substantial DPPH free radical-scavenging activity of the hydro ZnO/Se NPs. In vitro analyses demonstrated that hydro ZnO/Se NPs significantly inhibited cell proliferation, reduced migration, and diminished colony formation, likely due to the upregulation of CASP3, CASP8, and CASP9 gene expression. These findings highlight the therapeutic potential of hydro ZnO/Se NPs as promising anticancer agents for cervical cancer
