With the rapid development of nanotechnology, much has been anticipated with copper oxide nanoparticles (CuO NP) due to their extensive industrial and commercial application. However, it has raised concern over the environmental safety and human health effects. In this study, CuO nanoparticles were synthesized using the green method with floral extract of Calotropis gigantea and characterized by standard physiochemical techniques like DLS, Zeta potential determination, UV- Visible Spectroscopy, XRD, FTIR, FESEM, and TEM. Mechanistic cytotoxicity studies were performed using experimental and computational assays including morphological analysis, hatching, and viability rate analysis along with ROS and apoptosis analysis. Physiochemical characterization of CuO NP determined the size and zeta potential of synthesized nanoparticles to be 30 ± 09 nm to 40 ± 2 nm and −38 mV ± 12 mV respectively. Cytotoxicity evaluation with Zebrafish revealed malfunctioned organ development with differential viability and hatching rate at 48 hpf and 72 hpf with LC50 of 175 ± 10 mg/l. Computational analysis depicted the influential role of CuO nanoparticles on zebrafish embryo’s he1a, sod1 and p53 functional expression through hydrophobic and hydrogen bond interaction with amino acid residues. Study demonstrated valuable information of cytotoxic impact which can be influential in further studies of their eco-toxicological effects.

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Aim: To investigate the biocompatibility of green synthesized copper oxide nanoparticles (CuO Np) using floral extract of Calotropis gigantea in room condition. Materials & methods: Green synthesized and characterized CuO Np was evaluated for their cellular and molecular biocompatibility by experimentally and computational molecular docking. Results: Synthesized CuO NP was found to have a size 32 ± 09 nm with ζ potential -35 ± 12 mV. LC50 value was found to be 190 μg/ml. In vitro and in silico cytotoxicity analysis with HEK293 cells revealed the cytotoxic effect of CuO Np as consequences of interaction with histidine and arginine amino acid residues of Sod3 and p53 proteins via hydrogen bond of length 3.09 and 3.32 Å leading to oxidative stress ensuing toward apoptosis and cell cycle arrest. Conclusion: The outcomes proved the synthesized material as an alternative to the conventional method of synthesizing copper nanoparticles for biomedical and clinical applications.

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