Using a two-step method of surface mechanical attrition treatment and calcium ion implantation to promote the osteogenic activity of mesenchymal stem cells as well as biomineralization on a β-titanium surface

Surface treatment is known as a very efficient measure by which to modulate the surface properties of biomaterials in terms of grain structure, topography, roughness and chemistry to determine the osseointegration of implants. In this work, a two-step method of surface modification was employed to impart high osteogenic activity and biomineralization capacity on a Ti–25Nb–3Mo–2Sn–3Zr alloy (a type of β-titanium named TLM). The preliminary surface mechanical attrition treatment (SMAT) refined the average grain size from 170 ± 19 μm to 74 ± 8 nm in the TLM surface layer and promoted the surface to be much rougher and more hydrophilic. The subsequent Ca-ion implantation did not change the surface roughness and topography obviously, but enhanced the surface wettability of the SMAT-treated TLM alloy. The in vitro evaluations of the adhesion, proliferation, osteogenic genes (RUNX2, ALP, BMP-2, OPN, OCN and COL-I) and protein (ALP, OPN, OCN and COL-I) expressions, as well as extracellular matrix (ECM) mineralization of mesenchymal stem cells (MSCs) revealed that the initial SMAT-treated sample significantly enhanced the adhesion and osteogenic functions of MSCs compared to an untreated TLM sample, and the subsequent introduction of Ca ions onto the SMAT-derived nanograined sample further promotes the MSC adhesion, proliferation, osteo-differentiation and ECM mineralization due to the adsorption of more proteins such as laminin (Ln), fibronectin (Fn) and vitronectin (Vn) on the surface, as well as the increase in extracellular Ca concentrations. In addition, the biomineralization capacity of the samples was also evaluated by soaking them in simulated bodily fluid (SBF) at 37 °C for 28 days, and the results showed that the Ca-ion implanted sample significantly boosted the deposition of Ca and P containing minerals on its surface, which was associated with the generation of more Ti–OH groups on the surface after ion implantation. The combination of the SMAT technique and Ca-ion implantation thus endowed the TLM alloy with outstanding osteogenic and biomineralization properties, providing a potential means for its future use in the orthopedic field.