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Fullerene-titanium (C60/Ti) composites cause no DNA damage response in human osteoblast-like MG 63 cells

Kopova I., Bacakova L., Vacik J., Lavrentiev V.

Engineering of Biomaterials

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2010

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Vol. 13, no. 99-101

109--110

EN

Fullerenes (C60) and fullerene-based composites are considered as promising substrates for biological cell colonization. It might be mainly due to their nanostructure, resembling the nanoarchitecture of the natural extracellular matrix. Thin films of binary C60/Ti composites with various concentrations of Ti ranging from 25% (i.e., 25 Ti atoms and 75 C60 molecules) to 70% were deposited on microscopic glass coverslips in micro-patterned form through a metallic mask, and were tested for their potential use in bone tissue engineering. It is known that fullerenes and their derivatives can cause cytotoxic injury, cell death or inhibition of cell growth. These effects are based mainly on the reactivity of fullerenes, which may weaken with time due to the oxidization and polymerization of fullerenes in an air atmosphere. We therefore tested the dependence between the age of C60/Ti composites (i.e., from one week to one year) and the level of DNA damage of human osteoblast-like MG 63 cells in cultures on these materials. The DNA damage was analyzed by immunofluorescence staining of markers of DNA damage response, such as phosphorylation of histone H2AX and focal recruitment of p53-binding protein. As positive control to markers of DNA damage response was used 7 days long treatment with 2,5 mM Thymidine. We also monitored the proliferation and morphological changes of the cells. After 7 days of cultivation, we observe no cytotoxic morphological changes, such as enlarged cells or cytosolic vacuole formation, which are signs of cell senescence, and no increased induction of cell death. In addition, there was no increased level of DNA damage response on the C60/Ti composites (FIG.1). We also found no significant differences in cell population densities and no increased level of DNA damage among various Ti concentrations (FIG.1). Moreover, there was no effect of the age of the C60/Ti composites on the cell population densities or on the DNA damage response (FIG.1). These results suggest that fullerenes in combination with Ti do not cause cytotoxic injury and this material could be used in bone tissue engineering.

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Boron-doped nanocrystalline diamond films as substrates for adhesion, growth and differentiation of human osteoblast-like cells

Bacakova L., Grausova L., Kromka A., Vacik J., Lisa V., Rezek B., Haenen K.

Engineering of Biomaterials

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2010

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Vol. 13, no. 96-98

138--139

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Micro- and nanopatterned surfaces for guided adhesion, growth and phenotypic maturation of cells

Bacakova L., Filova E., Grausova L., Vandrovcova M., Parizek M., Novotna K., Svorcik V., Vacik J., Rypacek F., Kromka A., Heitz J., Shard A.

Engineering of Biomaterials

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2009

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Vol. 12, no. 89-91

18--21

EN

Micropatterned surfaces were created by UV light-irradiation of polytetrafluoroethylene through a metallic mask, by successive plasma polymerization of acrylic acid and 1,7-octadiene, or by creation of prominences and grooves by deposition of fullerenes C60 through a metallic mask. All these surface types were capable of inducing regionally-selective adhesion, proliferation and phenotypic maturation of vascular endothelial cells, vascular smooth muscle cells or human bone-derived MG 63 cells. Nanopatterned surfaces created by tethering GRGDSG oligopeptides through polyethylene oxide chains on a polymeric surface promoted spreading, formation of focal adhesion plaques and DNA synthesis in vascular smooth muscle cells. Surfaces nanopatterned with nanocrystalline diamond gave good support for the adhesion, growth and metabolic activity of osteoblast-like MG 63 cells.

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Adhesion of bone and vascular cells on carbon fibre-reinforced carbon composites coated with a fullerene layer

Bacakova L., Grausova L., Vacik J., Jungova I.

Inżynieria Biomateriałów

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2005

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R. 8, nr 47-53

3--6

EN

Carbon fibre-reinforced carbon composites (CFRC), i.e. materials promising for hard tissue surgery, were coated by a fullerene layer in order to strengthen the material surface and create its nanostructure pattern which is known to be attractive for colonization with bone cells. The fullerene layer was relatively resistant to wear, at least swabbing with cotton, rinsing with liquids and exposure to cells and proteolytic enzymes. Both human osteoblast-like MG 63 cells and rat vascular smooth muscle cells (VSMC) in 1-and 2-day-old cultures adhered to these surfaces in Iower numbers in comparison with the control uncoated material and tissue culture polystyrene. In addition, the VSMC on the fullerene-coated surfaces were less spread. The Iower cell adhesion was probably due to a relatively high hydrophobicity of fullerenes. On the other hand, the spreading of MG 63 cells was comparable to that observed on the control surfaces, and these cells also assembled dot-like vinculin-containing focal adhesion plaques and relatively rich fine filamentous beta-actin cytoskeleton. We suppose that the cell adhesion may be enhanced by derivatization of fullerenes with specific chemical functional groups or peptidic ligands for cell adhesion receptors.