Advances i n b iomaterial su rface s tructure a nd de sign h ave i mproved t issue eng ineering. We re port speci cally in this chapter on vertically aligned TiO2 nanotube surface structuring for the optimization of t itanium (Ti) i mplants ut ilizing na notechnology. e TiO2 na notube su rface s tructure formation, mechanism, biocharacteristics, and emerging role in tissue engineering and regenerative medicine are reviewed. e main focus will be on the unique 3-D tube-shaped nanostructure of TiO2 and its e ects on creating profound impacts on cell behavior. In particular, we discuss how the adhesion, proliferation, phenotypic f unctionality, a nd d i erentiation a re en hanced o n su rfaces w ith T iO2 na notube su rface structuring for advancing orthopedic and related applications. In fact, the presence of TiO2 nanotube surface structuring on Ti for orthopedic applications had a critical e ect that improved the proliferation and mineralization of osteoblasts in vitro, enhanced the bone bonding strength with rabbit t ibia over conventional Ti implants in vivo, and induced human mesenchymal stem cell (MSC) di erentiation into an osteogenic lineage because of the unique nanotopographical features and high-quality biocompatibility of the TiO2 nanotube surface. e nanotopography of the TiO2 nanotubular surface structure also has sub stantial i n uence o n up regulating e xtracellular m atrix ( ECM) p roduction i n c hondrocyte cultures for potential use in osteochondral bone/cartilage interface materials. is type of TiO2 surface nano-con guration is advantageous in regulating many positive cell and tissue responses for various tissue engineering and regenerative medicine applications.