Moravec Hynek, Vandrovcova Marta, Chotova Katerina, Fojt Jaroslav, Pruchova Eva, Joska Ludek, Bacakova Lucie
University of Chemistry and Technology, Prague, Department of Metals and Corrosion Engineering, Technicka 5, Prague 6, Czech Republic.
Department of Biomaterials and Tissue Engineering, Institute of Physiology of the Czech Academy of Sciences, Czech Republic.
Mater Sci Eng C Mater Biol Appl. 2016 Aug 1;65:313-22. doi: 10.1016/j.msec.2016.04.037. Epub 2016 Apr 13.
Nanotubes with diameters ranging from 40 to 60nm were prepared by electrochemical oxidation of the Ti-6Al-4V alloy in electrolyte containing ammonium sulphate and ammonium fluoride. The nanotubes were further modified with calcium and phosphate ions or were heat treated. Polished Ti-6Al-4V alloy served as a reference sample. The spreading of human osteoblast-like cells was similar on all nanotube samples but lower than on polished samples. The number of initially adhered cells was higher on non-modified nanotubes, but the final cell number was the highest on Ca-enriched nanotubes and the lowest on heat-treated nanotubes. However, these differences were relatively small and less pronounced than the differences in the concentration of specific molecular markers of cell adhesion and differentiation, estimated by their intensity of immunofluorescence staining. The concentration of vinculin, i.e. a protein of focal adhesion plaques, was the lowest on nanotubes modified with calcium. Collagen I, an early marker of osteogenic cell differentiation, was also the lowest on samples modified with calcium and was highest on polished samples. Alkaline phosphatase, a middle marker of osteogenic differentiation, was observed in lowest concentration on nanotubes modified with phosphorus and the highest on heat-treated samples. Osteocalcin concentrations, a late marker of osteogenic cell differentiation, were similar on all tested samples, although they tended to be the highest on heat-treated samples. Thus, osteogenic differentiation can be modulated by various additional treatments of nanotube coatings on Ti-6Al-4V implants.
通过在含有硫酸铵和氟化铵的电解质中对Ti-6Al-4V合金进行电化学氧化，制备了直径范围为40至60纳米的纳米管。这些纳米管进一步用钙离子和磷酸根离子进行了改性，或进行了热处理。抛光的Ti-6Al-4V合金用作参考样品。人成骨样细胞在所有纳米管样品上的铺展情况相似，但低于在抛光样品上的铺展情况。初始黏附细胞的数量在未改性的纳米管上较高，但最终细胞数量在富含钙的纳米管上最高，在热处理的纳米管上最低。然而，这些差异相对较小，且不如通过免疫荧光染色强度估计的细胞黏附与分化的特定分子标志物浓度差异明显。粘着斑蛋白（即一种粘着斑蛋白）的浓度在经钙改性的纳米管上最低。I型胶原蛋白是成骨细胞分化的早期标志物，在经钙改性的样品上也最低，在抛光样品上最高。碱性磷酸酶是成骨分化的中期标志物，在经磷改性的纳米管上浓度最低，在热处理的样品上浓度最高。骨钙素浓度是成骨细胞分化的晚期标志物，在所有测试样品上相似，尽管在热处理的样品上往往最高。因此，Ti-6Al-4V植入物上纳米管涂层的各种额外处理可以调节成骨分化。
