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Adhesion, growth and differentiation of osteoblast-like cells on materials for bone implants
Doubková, Martina ; Bačáková, Lucie (advisor) ; Filová, Eva (referee)
This thesis focuses on testing and improving Ti-6Al-4V ELI biomaterials, which are currently one of the most used titanium alloys in biomedicine (predominantly in orthopaedics and dentistry), in cooperation with research institutions and private companies developing and producing such materials. The metallic samples were previously modified by plasma electrolytic oxidation (PEO) with use of electrolytes of a different composition to induce development of a homogeneous TiO2 layer on its surface. In vitro interactions of human osteoblast-like cell line Saos-2 with the surface of Ti-6Al-4V ELI alloy samples are investigated. Initial cell attachment, spreading, morphology, cell population density, viability, calcium deposition and expression of selected osteogenic markers, e.g. collagen type I, alkaline phosphatase and osteocalcin, were evaluated on cultured cells. The cells behavior were then correlated with physicochemical properties of the material surface, such as its topography, roughness, wettability, surface layer chemical composition etc. The results are also compared with those obtained in cells cultured on control samples of untreated alloys as well as microscopic glass coverslips and bottom of standard polystyrene cell culture wells. The aim of this thesis is to select the most promising...
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Growth and differentiation of human osteoblast-like cells on TI-6AL-4V alloy modified with anodization
Doubková, Martina ; Bačáková, Lucie ; Pařízek, Martin ; Vandrovcová, Marta ; Musílková, Jana ; Lisá, Věra ; Gabor, R. ; Marvan, J.
Titanium and its alloys are widely used as hard tissue implants for their high biocompatibility and suitable physical properties. Along with other surface treatment methods, anodization technique is known to enhance osseointegration. The aim of this study is to evaluate the adhesion, growth and osteogenic differentiation of human osteoblast-like SAOS-2 cells on Ti-6Al-4V samples anodized in electrolytes composed of a stable volume of KOH (336.48 g/l) and variable volumes of liquid glass (124.47 g/l, 84,12 g/l and 37.38 g/l; samples s5, s6 and s7, respectively). Non-anodized Ti-6Al-4V samples, cell culture polystyrene (PS) and microscopic glass coverslips served as control materials. On days 2 and 4 after seeding, the cell number did not differ significantly among the tested samples. However, on day 7, the cell number on s6 samples reached the lowest values, which could be attributed to a non-homogeneous TiO2 film on s6 samples formed during anodization. Nevertheless, the osteogenic differentiation, estimated by the intensity of fluorescence of collagen I in cells grown in a differentiation medium, was the highest on s6 samples. On s5 samples, coated with homogeneous TiO2 films, both cell numbers and intensity of fluorescence of collagen I was relatively high. The bone matrix mineralization, evaluated by Alizarin Red staining, was the highest on s5 samples in standard culture medium, and similar on all tested samples in differentiation medium. Thus, the surface modification of s5 samples could be considered the most suitable for application in bone implants.
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