Bio-Synthesis of TiO and Ag-TiO Nanoparticles using Aegle marmelos Seed Extract and their Potential Efficacy Investigation in Human Leukemic Cell Proliferation, Inflammation, and Oxidative Stress.

The current study described the bio-functionalized metal nanoparticles TiO and Ag-TiO synthesis using Aegle marmelos (stone apple) seed extract. The TiO and Ag-TiO NPs were characterized by ultraviolet-visible (UV-Vis) spectroscopy, Fourier-transform infrared spectroscopy (FTIR), and scanning electron microscopy energy-dispersive X-ray (SEM-EDX) analysis. The stability of green-synthesized nanoparticles was studied by zeta potential analysis. Both NPs showed Cytotoxic effects on K562 (a cell line for human acute leukemia) with different efficacy IC value of TiO NPs was 151.3 µg/ml, whereas 27.65 µg/ml for Ag-TiO NPs. Microscopic analysis (100x) revealed concentration-dependent morphological changes, with Ag-TiO NPs exhibiting enhanced cytotoxicity. A flow cytometry study showed that there were significant increases in late apoptotic cells and decreases in healthy cells after treatment with Ag-TiO NPs. Overall these studies confirmed anti-cancer activity of TiO and Ag-TiO NPs. Free radical scavenging property was studied by DPPH assay, both TiO and Ag-TiO exhibits antioxidant activity with IC value of 437.1 μg/ml (Ag-TiO NP) and 544.1 μg/ml (TiO NP) respectively. This result predicts silver doping enhanced an antioxidant property. In vivo anti-inflammatory activity surprisingly showed TiO₂ and Ag-TiO₂ NPs dramatically suppress COX-2 and TNF-α production in lipopolysaccharide (LPS)-stimulated cells and also cut down expression of IL-1α and CRP levels in a dose-dependent manner. Furthermore, the same was confirmed by histopathological examination; TiO₂ and Ag-TiO₂ NPs mitigate tissue damage and inflammation and also showed hepatoprotective properties. In conclusion, both TiO and Ag-TiO NPs exhibit strong anti-cancer, antioxidant, and anti-inflammatory properties in both in vitro and in vivo conditions.