Selenium nanoparticle modified surfaces attract increasing attention in the field of tissue engineering. Selenium exhibits strong anticancer, antibacterial and anti-inflammatory properties and it maintains relatively low off-target cytotoxicity. In our paper, we present the fabrication, characterization and cytocompatibility of titanium oxide (TiO2) nanotube surface decorated with various surface densities of chemically synthesized selenium nanoparticles. To evaluate antibacterial and anti-cancer properties of such nanostructured surface, gram negative bacteria E. coli, cancerous osteoblast like MG-63 cells and non-cancerous fibroblast NIH/3T3 were cultured on designed surfaces. Our results suggested that selenium nanoparticles improved antibacterial properties of titanium dioxide nanotubes and confirmed the anticancer activity towards MG-63 cells, with increasing surface density of nanoparticles. Further, the selenium decorated TiO2 nanotubes suggested deteriorating effect on the cell adhesion and viability of non-cancerous NIH/3T3 cells. Thus, we demonstrated that selenium nanoparticles decorated TiO2 nanotubes synthesized using sodium selenite and glutathione can be used to control bacterial infections and prevent the growth of cancerous cells. However, the higher surface density of nanoparticles adsorbed on the surface was found to be cytotoxic for non-cancerous NIH/3T3 cells and thus it might complicate the integration of biomaterial into the host tissue. Therefore, an optimal surface density of selenium nanoparticles must be found to effectively kill bacteria and cancer cells, while remaining favorable for normal cells.