Titanium and its alloys are widely used for substitution of hard tissues, especially in orthopaedic and dental surgery. Despite the benefit of the use of titanium for such applications, there are still questions which must be sorted out. Surface properties are crucial for cell adhesion, proliferation and differentiation. Mainly, micro/nanostructured surfaces positively influence osteogenic differentiation of human mesenchymal stem cells. Ti6Al4V is a biocompatible α + β alloy which is widely used in orthopaedics. The aim of this study was to investigate the interaction of the nanostructured and ground Ti6Al4V titanium alloys with simulated body fluid complemented by the defined precipitation of hydroxyapatite-like coating and to study the cytotoxicity and differentiation capacity of cells with such a modified titanium alloy. Nanostructures were fabricated using electrochemical oxidation. Human mesenchymal stem cells (hMSC) were used to evaluate cell adhesion, metabolic activity and proliferation on the specimens. The differentiation potential of the samples was investigated using PCR and specific staining of osteogenic markers collagen type I and osteocalcin. Our results demonstrate that both pure Ti6Al4V, nanostructured samples, and hydroxyapatite-like coating supported hMSC growth and metabolic activity. Nanostructured samples improved collagen type I synthesis after 14 days, while both nanostructured and hydroxyapatite-like coated samples enhanced collagen synthesis on day 21. Osteocalcin synthesis was the most enhanced by hydroxyapatite-like coating on the nanostructured surfaces. Our results indicate that hydroxyapatite-like coating is a useful tool guiding hMSC osteogenic differentiation.

2nd Faculty of Medicine Charles University Prague 5 Úvalu 84 150 06 Prague Czech Republic

Department of Metals and Corrosion Engineering Faculty of Chemical Technology University of Chemistry and Technology Technická 5 166 28 Prague Czech Republic

Department of Tissue Engineering Institute of Experimental Medicine of the Czech Academy of Sciences Vídeňská 1083 Prague 4 142 20 Czech Republic

Faculty of Science Charles University Prague Albertov 2038 6 128 00 Prague Czech Republic

University Centre for Energy Efficient Buildings Czech Technical University Prague Třinecká 1024 273 43 Buštěhrad Czech Republic

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