Anti angiogenic and anticancer activities of nanoformulated naringenin with combination of selenium nanoparticles evaluated on in vitro and in ova models

Abstract

Cancer poses a substantial global health challenge, demanding innovative therapeutic approaches that can effectively target both tumor growth and progression. Among these approaches, angiogenesis, the process of generating new blood vessels from pre-existing ones, holds a pivotal role in various physiological and pathological processes, including tumor growth, wound healing, and inflammatory disorders. Targeting angiogenesis has emerged as a promising strategy for developing novel therapeutic interventions against a range of diseases, including cancer. In this study, we investigate the potential anti-cancer and anti-angiogenic properties of both individually and combination of nanoformulated naringenin (NF-n) and selenium nanoparticles (SeNPs). The synthesis of NF-n was achieved through the solvent precipitation method and its characterization was analysed by Dynamic Light Scattering (DLS), Fourier Transform Infrared Spectroscopy (FT-IR), and Scanning Electron Microscopy (SEM). Meanwhile, SeNPs were synthesized via the Glutathione reduction method, and their characterization included DLS and Transmission Electron Microscopy (TEM). For assessing the in vitro anti-angiogenic activity of NF-n and SeNPs, we used Human Umbilical Cord Vascular Endothelial Cells (HUVECs). This in vitro assessment involved analysing cell viability, cell migration, and the mRNA expressions of key angiogenic markers such as Nestin, Neuropilin-1 (NRP-1), Neuropilin-2 (NRP-2), and CD93. To evaluate in vitro anti-cancer activity, we used Michigan Cancer Foundation-7 (MCF-7) breast cancer cell lines and performed cell viability, cell migration and mRNA expressions such as TNF-and#945;, NF-and#954;B, and Bcl-2. The in ova anti-angiogenic effects were examined on the Chick Chorioallantoic Membrane (CAM) model, where quantitative measurement of blood vessel branches, histopathological examination of CAM, immunohistochemical analysis of CAM and mRNA expression analyses were conducted. newline The NF-n was successfully synthesized by solvent precipitation method and synthesized nanoparticles were characterized by DLS, FT-IR and SEM. The SeNPs were successfully synthesized and were characterized by DLS and TEM. The in vitro anti-angiogenic assays such as cell proliferation and cell migration results demonstrated combination of NF-n and SeNPs led to a significant inhibition of cell proliferation and migration of HUVEC compared to control upon with alone and combinational with NF-n and SeNPs treatments. The combination also resulted in a notable reduction in mRNA expression levels compared to the control and individual treatment with drugs (NF-n and SeNPs). The in vitro anti-cancer activity results exhibited significant inhibition of cell proliferation and migration in MCF-7 cell lines. The mRNA expressions of TNF-and#945;, NF-and#954;B, and Bcl-2 were significantly downregulated after the combinational treatment of NF-n and SeNPs compared to individual treatments. The Morphometric analysis of the in ova CAM model demonstrated that the combination of NF-n and SeNPs significantly decreased the size of blood vessels including vessel density, total nets, total branch points, and total vessel network length. Histopathological findings indicated thinner vessel walls and reduced numbers of newly formed blood vessels compared to the control group. Moreover, mRNA expression levels of key factors like Vascular Endothelial Growth Factor-A (VEGF-A), VEGF-R2, Ang-1, Ang-2 and Cyclophilin-A (Cyc-A) were significantly downregulated upon the combination treatment with NF-n and SeNPs. Overall, this study sheds light on the promising potential of combining NF-n and SeNPs to elicit potent anti-cancer and anti-angiogenic effects, providing a valuable foundation for the development of cancer therapies. newline newline