Natural bone consists of hard nanostructured hydroxyapatite (HA) in a nanostructured protein-based soft hydrogel template (ie, mostly collagen). For this reason, nanostructured HA has been an intriguing coating material on traditionally used titanium for improving orthopedic applications. In addition, helical rosette nanotubes (HRNs), newly developed materials which form through the self-assembly process of DNA base pair building blocks in body solutions, are soft nanotubes with a helical architecture that mimics natural collagen. Thus, the objective of this in vitro study was for the first time to combine the promising attributes of HRNs and nanocrystalline HA on titanium and assess osteoblast (bone-forming cell) functions. Different sizes of nanocrystalline HA were synthesized in this study through a wet chemical precipitation process following either hydrothermal treatment or sintering. Transmission electron microscopy images showed that HRNs aligned with nanocrystalline HA, which indicates a high affinity between both components. Some of the nanocrystalline HA formed dense coatings with HRNs on titanium. More importantly, results demonstrated enhanced osteoblast adhesion on the HRN/nanocrystalline HA-coated titanium compared with conventional uncoated titanium. Among all the HRN/nanocrystalline HA coatings tested, osteoblast adhesion was the greatest when HA nanometer particle size was the smallest. In this manner, this study demonstrated for the first time that biomimetic HRN/nanocrystalline HA coatings on titanium were cytocompatible for osteoblasts and, thus, should be further studied for improving orthopedic implants.