The effects of surface treatments on electron beam melted Ti-6Al-4V disks on osteogenesis of human mesenchymal stromal cells

Additive manufactured (AM) Titanium-6Aluminum-4Vanadium (Ti64) scaffolds display unique mechanical and biological properties for implant devices. The elastic modulus can be tailored by adjusting the porosity, further facilitating bone ingrowth. Although Ti64 implants are biocompatible, the effects of AM surfaces without porous structures, and how the topography and surface chemistry of the respective surfaces affect the osteogenesis of bone marrow-derived mesenchymal stromal cells (BMSCs) has not yet been revealed. In this paper, we cultured BMSCs on solid electron beam melted Ti64 disks subjected to three surface treatments: chemical etching (HF), atomic-layer deposition of TiO₂ (TiO₂), and polished (POL), or left untreated (AB). The biocompatibility and osteogenic properties of these surfaces were investigated, and the results were compared to cells cultured in regular tissue-culture polystyrene culturing wells (TCPS). The surfaces were hydrophobic, except for the polished surface which was hydrophilic. All surface treatments are biocompatible and allow for osteogenic differentiation, as revealed by viability assays and gene expression analysis. Scanning electron microscopy shows that cells adhere differently depending on the surface properties, with more filopodia on the rougher surfaces, AB and TiO₂ disks, and more lamellipodia on the smoother surfaces, HF and POL disks. All groups stimulated with beta glycerophosphate, ascorbic acid, and dexamethasone, have elevated expression of genes related to matrix formation, where the cells cultured on the disks treated with TiO₂, HF and POL have the overall highest expression. The AB group appears to be less favorable in regards to matrix formation. Considering the matrix mineralization, the rougher surfaces, AB and TiO₂, are able to induce matrix mineralization, with an elevated gene expression of vitamin D receptors and calcium deposition of unstimulated cells. Finally, imaging at day 21 revealed an even amount of cells and matrix, covering most of the partially melted particles. Our results suggests that surface topography is more important to osteogenesis than the wettability of the surface. Overall, the present study contributes to the understanding of using surface modifications to AM Ti64 implant materials and reveals how they affect bone growth.