A metal/peptide interface was developed to nucleate hydroxyapatite from aqueous solution. To achieve this, three different modifications were performed on mirror polished Ti-6Al-4V (MPT): 1) thermal oxidation (TO) at 500, 600, 700, 800 °C, 2) chemical treatment with H2O2/HCl and 3) chemical treatment with H2O2/HCl followed by TO at 500, 600, 700, 800 °C. It was found that with the increase of the TO temperature the topography increased and the composition of the surfaces changed, forming TiO2 crystals and increasing the Al2O3 content on the surface. These changes affected locally the surfaces charge behaviour in acidic and alkaline aqueous environments that were clue to the electrostatic interaction between the peptide, which presented positive and negative charged amino acids, and the surface. The peptide adsorption was highest on the surfaces that was chemically treated and thermal oxidised at 500 °C. The peptide coating was confirmed on the surface by the change of the charge behaviour in acidic and alkaline aqueous environments and its chemical composition. In addition, the growth of hydroxyapatite (HA) was obtained on the peptide coated surfaces confirming its bioactivity. Finally, it was shown that the surfaces with HA presented a better cell adhesion than the other modified surfaces.