Nanotopography has a significant regulatory effect on cell adhesion behavior. We fabricated titanium alloy surfaces with titanium dioxide nanotubes using electrochemical anodization, which exhibited increased surface roughness and superhydrophilicity. Using single-cell force spectroscopy, we observed that the nanotubular structure altered the mechanical forces experienced at the cell-surface interface. Proteomic analysis revealed that Nesprin-1 protein transmits these signals to the intracellular core signaling system, leading to the upregulation and activation of GSK3β, STAT3, and related pathways. These events regulate cytoskeletal remodeling and adhesion complex assembly, thereby promoting cell adhesion.