The research's goal is to create the surfaces of titanium dioxide nanoparticles (TiO2 NPs) in a layer of folic acid (FA) that can effectively target human bladder cancer cells (T24). An efficientmethod for creating FA-coated TiO2 NPs was used, and many tools have been used to analyze its physicochemical properties. The cytotoxic effects of FA-coated NPs on T24 cells and the mechanisms of apoptosis generation were examined employing a variety of methodologies. The prepared FA-coated TiO2 NPs suspensions with a hydrodynamic diameter around 37nm and a negative surface charge of -30mV reduced T24 cell proliferation with stronger IC50 value (21.8±1.9 μg/ml) than TiO2 NPs (47.8±2.5 μg/ml). This toxicity resulted in apoptosis induction (16.63%) that was caused through enhanced reactive oxygen species formation and stopping the cell cycle over G2/M phase. Moreover, FA-TiO2 NPs raised the expression levels of P53, P21, BCL2L4, and cleaved Caspase-3, while decreasing Bcl-2, Cyclin B, and CDK1 in treated cells. Overall, these findings revealed efficient targeting of the FA-TiO2 NPs resulted in increasing cellular internalization caused increased apoptosis in T24 cells. As a result, FA-TiO2 NPs might be a viable treatment for human bladder cancer.