Biosynthesis and Characterization of Zinc Oxide‐Doped Selenium Nanocomposite Conjugated With <scp>l</scp> ‐Asparaginase Against Human Hepatoma Cell Line

Nanocarriers have emerged as promising systems for improving drug-delivery efficiency and advancing diagnostic strategies in cancer treatment. l-asparaginase has well-established antitumor and anti-leukemic properties, but its therapeutic performance is often restricted due to its short biological half-life and poor instability in the unmodified form. The present study investigated the encapsulation of l-asparaginase on a green-synthesized nanocarrier to enhance its overall therapeutic efficacy. The nanocarrier was produced utilizing citrus orange peel extract, which served simultaneously as a reducing, stabilizing, and capping material during the formation of zinc oxide-doped selenium nanoparticles (ZnO-Se-NPs). This eco-friendly synthesis method not only stabilized the nanoparticles but also incorporated naturally occurring phytochemicals, including flavonoids, phenolic compounds, ascorbic acid, and carotenoids-that may contribute to additional pharmacological benefits. The synthesized l-asparaginase encapsulated ZnO-Se-NPs (ZnO-Se-Asp) nanocomposite exhibited notable anticancer activity while maintaining low cytotoxicity. The detailed characterization of the nanoparticles was carried out utilizing UV-Vis spectroscopy to confirm the presence of ZnO-Se-Asp, energy dispersive x-ray (EDX), and scanning electron microscope (SEM) for morphological and elemental analysis. The Fourier transform infrared spectroscopy (FTIR) was studied to identify the functional groups, and x-ray diffraction (XRD) was performed to study the crystallinity of synthesized ZnO-Se-Asp nanocomposite. The cytotoxic potential of synthesized ZnO-Se-Asp nanocomposite was estimated using the methyl thiazolyl tetrazolium (MTT) assay through the determined IC₅₀ value, as well as its effects on the cell viability and proliferation were assessed. Finally, intracellular reactive oxygen species (ROS) levels and Caspase-3 activity were performed to determine oxidative stress generation and apoptotic induction. Overall, the results indicated that the green-synthesized ZnO-Se-Asp nanocomposite offers a promising platform capable of improving anticancer efficacy while offering better stability and bioavailability.