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The diameter of nanotubes formed on Ti-6Al-4V alloy controls the adhesion and differentiation of Saos-2 cells.

Filova E, Fojt J, Kryslova M, Moravec H, Joska L, Bacakova L - Int J Nanomedicine (2015)

Bottom Line: On day 3, the highest concentrations of both vinculin and talin measured by enzyme-linked immunosorbent assay and intensity of immunofluorescence staining were on 30 V nanotubes.On the other hand, the highest concentrations of ALP, type I collagen, and osteopontin were found on 10 V and 20 V samples.Therefore, the controlled anodization of Ti-6Al-4V seems to be a useful tool for preparing nanostructured materials with desirable biological properties.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomaterials and Tissue Engineering, Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.

ABSTRACT
Ti-6Al-4V-based nanotubes were prepared on a Ti-6Al-4V surface by anodic oxidation on 10 V, 20 V, and 30 V samples. The 10 V, 20 V, and 30 V samples and a control smooth Ti-6Al-4V sample were evaluated in terms of their chemical composition, diameter distribution, and cellular response. The surfaces of the 10 V, 20 V, and 30 V samples consisted of nanotubes of a relatively wide range of diameters that increased with the voltage. Saos-2 cells had a similar initial adhesion on all nanotube samples to the control Ti-6Al-4V sample, but it was lower than on glass. On day 3, the highest concentrations of both vinculin and talin measured by enzyme-linked immunosorbent assay and intensity of immunofluorescence staining were on 30 V nanotubes. On the other hand, the highest concentrations of ALP, type I collagen, and osteopontin were found on 10 V and 20 V samples. The final cellular densities on 10 V, 20 V, and 30 V samples were higher than on glass. Therefore, the controlled anodization of Ti-6Al-4V seems to be a useful tool for preparing nanostructured materials with desirable biological properties.

No MeSH data available.


Related in: MedlinePlus

Immunofluorescence staining of vinculin (green) and F-actin (red) in human Saos-2 osteoblasts.Notes: 10 V (A), 20 V (B), and 30 V (C) nanotubes, on control Ti_C (D), and on glass coverslips (E) on day 3. Cell nuclei (blue) were counterstained with Hoechst 33258. Leica SPE confocal microscope, objective 63×, zoom 2×, scale 25 μm.
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f6-ijn-10-7145: Immunofluorescence staining of vinculin (green) and F-actin (red) in human Saos-2 osteoblasts.Notes: 10 V (A), 20 V (B), and 30 V (C) nanotubes, on control Ti_C (D), and on glass coverslips (E) on day 3. Cell nuclei (blue) were counterstained with Hoechst 33258. Leica SPE confocal microscope, objective 63×, zoom 2×, scale 25 μm.

Mentions: Unfortunately, anodization does not allow for the preparation of nanotubes with a narrow size distribution. Nevertheless, we obtained similar cell-population densities on all nanotube layers, although they displayed a wide range of nanotube diameters (Figure 5). This can be explained by the fact that the wall thickness of all nanotubes (ie, 14±2 nm for 10 V, 18±4 nm for 20 V and 19±4 nm for 30 V nanotubes) was appropriate for integrin-mediated cell adhesion and the formation of focal adhesion plaques. The surface occupied by the head of the integrin receptor is approximately 10 nm, and the optimum space for focal adhesion plaques could be approximately 15 nm.46 In accordance with this, the cells on all tested nanotube films displayed well-developed focal adhesion plaques (Figure 6). Although the cells on the nanotube layers adhered at lower initial cell-population densities than on glass, they grew relatively quickly, and on day 7 they reached similar or even higher population densities (Figure 5).


The diameter of nanotubes formed on Ti-6Al-4V alloy controls the adhesion and differentiation of Saos-2 cells.

Filova E, Fojt J, Kryslova M, Moravec H, Joska L, Bacakova L - Int J Nanomedicine (2015)

Immunofluorescence staining of vinculin (green) and F-actin (red) in human Saos-2 osteoblasts.Notes: 10 V (A), 20 V (B), and 30 V (C) nanotubes, on control Ti_C (D), and on glass coverslips (E) on day 3. Cell nuclei (blue) were counterstained with Hoechst 33258. Leica SPE confocal microscope, objective 63×, zoom 2×, scale 25 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664495&req=5

f6-ijn-10-7145: Immunofluorescence staining of vinculin (green) and F-actin (red) in human Saos-2 osteoblasts.Notes: 10 V (A), 20 V (B), and 30 V (C) nanotubes, on control Ti_C (D), and on glass coverslips (E) on day 3. Cell nuclei (blue) were counterstained with Hoechst 33258. Leica SPE confocal microscope, objective 63×, zoom 2×, scale 25 μm.
Mentions: Unfortunately, anodization does not allow for the preparation of nanotubes with a narrow size distribution. Nevertheless, we obtained similar cell-population densities on all nanotube layers, although they displayed a wide range of nanotube diameters (Figure 5). This can be explained by the fact that the wall thickness of all nanotubes (ie, 14±2 nm for 10 V, 18±4 nm for 20 V and 19±4 nm for 30 V nanotubes) was appropriate for integrin-mediated cell adhesion and the formation of focal adhesion plaques. The surface occupied by the head of the integrin receptor is approximately 10 nm, and the optimum space for focal adhesion plaques could be approximately 15 nm.46 In accordance with this, the cells on all tested nanotube films displayed well-developed focal adhesion plaques (Figure 6). Although the cells on the nanotube layers adhered at lower initial cell-population densities than on glass, they grew relatively quickly, and on day 7 they reached similar or even higher population densities (Figure 5).

Bottom Line: On day 3, the highest concentrations of both vinculin and talin measured by enzyme-linked immunosorbent assay and intensity of immunofluorescence staining were on 30 V nanotubes.On the other hand, the highest concentrations of ALP, type I collagen, and osteopontin were found on 10 V and 20 V samples.Therefore, the controlled anodization of Ti-6Al-4V seems to be a useful tool for preparing nanostructured materials with desirable biological properties.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomaterials and Tissue Engineering, Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.

ABSTRACT
Ti-6Al-4V-based nanotubes were prepared on a Ti-6Al-4V surface by anodic oxidation on 10 V, 20 V, and 30 V samples. The 10 V, 20 V, and 30 V samples and a control smooth Ti-6Al-4V sample were evaluated in terms of their chemical composition, diameter distribution, and cellular response. The surfaces of the 10 V, 20 V, and 30 V samples consisted of nanotubes of a relatively wide range of diameters that increased with the voltage. Saos-2 cells had a similar initial adhesion on all nanotube samples to the control Ti-6Al-4V sample, but it was lower than on glass. On day 3, the highest concentrations of both vinculin and talin measured by enzyme-linked immunosorbent assay and intensity of immunofluorescence staining were on 30 V nanotubes. On the other hand, the highest concentrations of ALP, type I collagen, and osteopontin were found on 10 V and 20 V samples. The final cellular densities on 10 V, 20 V, and 30 V samples were higher than on glass. Therefore, the controlled anodization of Ti-6Al-4V seems to be a useful tool for preparing nanostructured materials with desirable biological properties.

No MeSH data available.


Related in: MedlinePlus