There are serious concerns on the toxicity of vanadium in Ti-6Al-4V alloy. In this regard, we describe the biological footprint of Ti-6Al-4V and compare with a viable alternate Ti-6Al-7Nb alloy, in terms of novel experimentation pertaining to cellular activity that include qualitative and quantitative analysis of Feret's diameter of cells, area, and perimeter, and proteins—actin, vinculin, and fibronectin. Interestingly, Ti-6Al-7Nb was characterized by superior cell attachment, proliferation, viability, morphology, and spread, which were significantly different from Ti-6Al-4V alloy. Additionally, immunofluorescence studies demonstrated stronger vinculin signals associated with actin stress fibers in the outer regions of the cells and cellular extensions in Ti-6Al-7Nb alloy. These striking observations suggest enhanced cell–substrate interaction and activity on the surface of niobium-containing titanium alloy. The significant differences in the cellular response between the two alloys clearly point to the determining role of alloying element (Nb versus V) in a conclusive manner. Based on this study, next generation of titanium alloys is proposed to focus on niobium-containing alloy. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013.