The main aim of this study was to investigate the effect of plastic deformation on the biological behaviour of novel Ti-29Nb-13Ta-4.6Zr alloy and to compare it with microcrystalline α titanium, commonly used in medicine. A multiple-stage cold rolling process was applied to improve both the mechanical and functional properties in a TNTZ alloy. A systematic characterization of the microstructure of all tested materials was performed, followed by a correlation with the mechanical properties and phase composition. A cellular response test based on osteosarcoma MG63 osteoblast like cells was carried out in order to analyse the influence of plastic deformation on the biocompatibility of the TNTZ alloy subjected to cold rolling. A detailed surface characterization including inter alia: contact angle and surface free energy, XPS, AES and AFM, was made to define the key factors affecting cellular adhesion to the surface of the deformed samples. The adhered cells were analyzed using confocal and atom force microscopies after 4 days of incubation. Our initial tests indicated that the novel β-Ti alloy subjected to the cold rolling process could be a promising new candidate in the fields of orthopaedics and dentistry implantations.