Effects of Acid-alkali Treatment on Bioactivity and Osteoinduction of Porous Titanium: An in Vitro Study

Background: To elucidate the bioactivity and bone regeneration of porous titanium surfaces treated using acid-alkali combination, and to define the optimal alkali reaction time.

Methods: Ten groups of porous Ti with at least 3 per group undergoing different acid-alkali treated time were prepared. The surface was characterized by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), bicinchoninic acid method (BCA), optical contact angle measurement and Raman spectrometry. Compression testing was performed with a universal testing machine. The bioactivity and osteoinduction were evaluated by a series of biological tests using a simulated body fluid (SBF) test, cell proliferation test, vinculin, ALP and OCN expression, and cell mineralization.

Results: The acid-alkali treatment formed micro- and nano-scale structures on the sample surfaces. The alkali treatment for 12 h achieved the sharpest nano-scale surface relief and the most protein absorption. The treated porous surface was coated with a NaHTiO layer. The acid-alkali etching did not compromise the elastic modulus and compressive strength of the porous Ti samples. In addition to hydroxyapatite, a perovskite phase was also formed on the treated porous samples in SBF. Non-treated dense Ti showed more cell adhesion and proliferation (P < 0.05), while osteoinduction and mineralization were more pronounced on the treated porous sample (P < 0.05).

Conclusion: Acid-alkali treatment is an effective means of generating nano-scale relief on porous Ti surface, and is beneficial for bioactivity and bone regeneration. The 15 min acid and 12 h alkali etching is the optimal combination. The osteoinductive efficacy may be attributable to the surface physical chemistry and the formation of hydroxyapatite and perovskite layers, rather than direct cell adhesion and proliferation.