Title: Phytofabrication of silver and cupric oxide nanoparticles using Simarouba glauca and Celastrus paniculatus extracts and their enhanced apoptotic inducing and anticancer effects

Abstract

Objectives and scope : Silver (AgNPs) and Cupric oxide nanoparticles (CuONPs) were phytofabricated utilizing leaf extract of Simarouba glauca (SG) and aerial extract of Celastrus paniculatus (CP) to evaluate anticancer effect and also to verify the apoptosis, cell cycle analysis. Methods used: free radical scavenging assays like DPPH, ABTS and NO; MTT assay, flow cytometry and caspase-3; EAC model with biochemical and haematological parameters. Results and discussions: Characterization was validated using FTIR, SEM-EDX, TEM, XRD and UV-Vis analysis. The green synthesized AgNPs and CuONPs showed potent antioxidant potential with IC50 value of about 34.01+0.64 µg/mL contrasted to ascorbic acid. The anticancer activities on cancerous cell lines like MCF-7 and HT-29 cell lines revealed that AgNPs and CuONPs synthesized using S. glauca and C. paniculatus indicated IC50 values ranging from 70.85+0.67 to 240.6+0.57 µg/mL. They could not effectively prohibit the growth of immortalized normal human breast epithelial cell lines (MCF-10A). To be more precise for anticancerous effect, molecular mechanism was examined in MCF-7 cell line treated with CuO-CP NPs by cell cycle analysis that depicted 75.28 % of cell arrest in Sub G0/G1 phase and 71.29 % of cells were gated in late apoptotic phase of Annexin V and propidium iodide (PI) compared to control cells. The cytotoxicity induced by apoptosis was further confirmed by fluorescent images. The synthesized nanoparticles also demonstrated less hemolysis efficiency and are evidenced by SEM images. We have also evaluated the in vivo antitumor efficacy of CuO-CP NPs treated against Ehrlich ascites carcinoma (EAC) bearing C57 mice for the first time and examined by variations in growth parameters, biochemical assays (like lactoperoxidase, reduced glutathione and myeloperoxidase), hematological profile, and histopathological analysis in comparison with control. Conclusion: The green synthesized nanoparticles exhibited effective control of cancer cells in both in vitro and in vivo laboratory conditions and thus can be evaluated for lead compound and scale up for industrial usage.