Studying the Efficacy of Copper Oxide Loaded Graphitic Carbon Nitride Nanosheets on VEGF-Induced Angiogenesis in Chick Chorioallantoic Membrane (CAM Assay) In Vivo Studies and Cytotoxicity in Human Hep3B Liver Cancer Cell Lines

Angiogenesis, a hallmark in many cancers, is the uncontrollable formation of blood vessels from the pre-existing vessels and leads to metastasis. The objective of the study was to first time evaluate the influence of CuO-loaded g-C 3 N 4 nanosheets to test its anti-angiogenic activity in the chorioallantoic membrane of chick and viability test in liver cancer cell line. The prepared material was tested using anti-cancer activity in vivo and in vitro analysis. Initially, prepared sphere-shaped-cupric oxide and cupric oxide supported g-C 3 N 4 nanosheet crystallinity, purity, morphology, and physicochemical properties (such as optical and thermal) are analyzed. On the other hand, the prepared materials angiogenesis analysis is examined using a series of chorioallantoic membrane (CAM) assays in the in vivo models which confirmed that the cupric oxide loaded on exfoliated g-C 3 N 4 nanosheets can eradicate the tumor cell. Results also showed that CuO/g-C 3 N 4 nanosheets strongly disturbed the physiological process of angiogenesis in the chorioallantoic membrane blocking the VEGF-induced blood vessel formation. The findings indicate that 40wt% of CuO/g-C 3 N 4 nanocomposite is effective and probably acts as anti-angiogenic therapy compared to other weight percentages of nanocomposite for the suppression of VEGF growth factor activities. Furthermore, the effect on cell viability is analyzed by using a human Hep3B liver cancer cell line when using the 40wt% CuO/g-C 3 N 4 nanocomposite. Graphical Abstract