Objectives: Nanotechnology holds an important area in recent research due of its immense use in the different field of sciences. The small size, large surface area, orientation, and physical properties make them appropriate to be used in medical sciences, physics, and chemistry. Physical and chemical processes of synthesizing nanoparticles are continued for ages. The biological approaches to preparing nanoparticles have drawn the attention of researchers due to eco-friendly nature, low cost, and easier steps for synthesis. To synthesize titanium nanoparticles, Vigna radiata legumes were powdered and used, and the effectiveness of biologically synthesized titanium nanoparticles against clinical pathogens and anticancer activity were checked. Methods: Green synthesis of titanium dioxide nanoparticles is carried out in simple steps. The extract of V. radiata legumes was used for the biological synthesis of the titanium dioxide nanoparticles which was characterized by Fourier transform infrared spectroscopy and scanning electron microscopy. Antibacterial activity of the titanium dioxide nanoparticles was checked against nine clinical pathogens. 2,2-diphenyl-1-picryl-hydrazyl-hydrate assay was performed to determine the antioxidant activity of the nanoparticles. Cytotoxic activity against osteosarcoma cell lines was performed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: Oval shaped titanium nanoparticles were biologically synthesized. The particles showed effectiveness against both Gram-positive and Gram-negative bacteria. The particles exhibited antioxidant nature. Cytotoxicity assay revealed the nanoparticles were capable of inhibiting proliferation of osteosarcoma cell lines. Conclusions: V. radiata has been confirmed to be a good source for biosynthesis of titanium nanoparticles. The cytotoxicity effect of the nanoparticles hints that it can be further used for treatment and medicine purposes. © 2016, Innovare Academics Sciences Pvt. Ltd. All rights reserved.