Tuning Mesenchymal Stem Cell Response onto Titanium-Niobium-Hafnium Alloy by Recombinant Fibronectin Fragments.

Since metallic biomaterials used for bone replacement possess low bioactivity, the use of cell adhesive moieties is a common strategy to improve cellular response onto these surfaces. In recent years, the use of recombinant proteins has emerged as an alternative to native proteins and short peptides owing to the fact that they retain the biological potency of native proteins, while improving their stability. In the present study, we investigated the biological effect of two different recombinant fragments of fibronectin, spanning the 8-10th and 12-14th type III repeats, covalently attached to a new TiNbHf alloy using APTES silanization. The fragments were studied separately and mixed at different concentrations and compared to a linear RGD, a cyclic RGD and the full-length fibronectin protein. Cell culture studies using rat mesenchymal stem cells demonstrated that low to medium concentrations (30% and 50%) of type III 8-10th fragment mixed with type III 12-14th fragment stimulated cell spreading and proliferation compared to RGD peptides and the fragments separately. On the other hand, type III 12-14th fragment alone or mixed at low volume percentages ≤50% with type III 8-10th fragment increased alkaline phosphatase levels compared to the other molecules. These results are significant for the understanding of the role of fibronectin recombinant fragments in cell responses and thus to design bioactive coatings for biomedical applications.

Bioactive compounds immobilized on Ti and TiNbHf: AFM-based investigations of biofunctionalization efficiency and cell adhesion.

Implant materials require optimal biointegration, including strong and stable cell-material interactions from the early stages of implantation. Ti-based alloys with low elastic modulus are attracting a lot of interest for avoiding stress shielding, but their osseointegration potential is still very low. In this study, we report on how cell adhesion is influenced by linear RGD, cyclic RGD, and recombinant fibronectin fragment III8-10 coated on titanium versus a novel low-modulus TiNbHf alloy. The bioactive molecules were either physisorbed or covalently coupled to the substrates and their conformation on the surfaces was investigated with atomic force microscopy (AFM). The influence of the different bioactive coatings on the adhesion of rat mesenchymal stem cells was evaluated using cell culture assays and quantitatively analyzed at the single cell level by AFM-based single-cell force spectroscopy. Our results show that bioactive moieties, particularly fibronectin fragment III8-10, improve cell adhesion on titanium and TiNbHf and that the covalent tethering of such molecules provides the most promising strategy to biofunctionalize these materials. Therefore, the use of recombinant protein fragments is of high importance for improving the osseointegration potential of implant materials.

Mechanical and physicochemical characterization along with biological interactions of a new Ti25Nb21Hf alloy for bone tissue engineering.

Nowadays, one of the main challenges in metal implants for bone substitution is the achievement of an elastic modulus close to that of human cortical bone as well as to provide an adequate interaction with the surrounding tissue avoiding in vivo foreign body reaction. From this perspective, a new Ti-based alloy has been developed with Nb and Hf as alloying elements which are known as non-toxic and with good corrosion properties. The microstructure, mechanical behaviour and the physicochemical properties of this novel titanium alloy have been studied. Relationship of surface chemistry and surface electric charge with protein adsorption and cell adhesion has been evaluated due to its role for understanding the mechanism of biological interactions with tissues. The Ti25Nb21Hf alloy presented a lower elastic modulus than commercial alloys with a superior ultimate strength and yield strength than CP-Ti and very close to Ti6Al4V. It also exhibited good corrosion resistance. Furthermore, the results revealed that it had no cytotoxic effect on rat mesenchymal stem cells and allowed protein adsorption and cell adhesion. The experimental results make this alloy a promising material for bone substitution or for biomedical devices.