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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

Human Saos-2 osteoblasts on 10 V, 20 V, and 30 V nanotubes, on control Ti_C, and on glass coverslips on day 3.Notes: Immunofluorescence intensity (A, B) and absorbance measured by enzyme-linked immunosorbent assay (C, D) of talin (A, C) and vinculin (B, D). Data expressed as mean ± standard error of mean. P≤0.05 considered significant in comparison with samples labeled above columns.
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f7-ijn-10-7145: Human Saos-2 osteoblasts on 10 V, 20 V, and 30 V nanotubes, on control Ti_C, and on glass coverslips on day 3.Notes: Immunofluorescence intensity (A, B) and absorbance measured by enzyme-linked immunosorbent assay (C, D) of talin (A, C) and vinculin (B, D). Data expressed as mean ± standard error of mean. P≤0.05 considered significant in comparison with samples labeled above columns.

Mentions: The concentration and distribution molecular markers of cell adhesion, ie, talin and vinculin, were evaluated by immunofluorescence staining and enzyme-linked immunosorbent assay (ELISA). The immunofluorescence of talin was relatively diffused, and focal adhesions were not clearly visible (Figure S1). The fluorescence intensity of talin per cell was most intense on the 30 V nanotube sample, and there was a very low signal on glass (Figure 7A). As mentioned earlier, well-developed focal adhesion plaques containing vinculin were found on all surfaces (Figure 6). They seemed to be slightly larger on smooth Ti_C surfaces than on nanotubes. On glass, focal adhesions in Saos-2 cells were smaller than on the other surfaces, and the signal of vinculin was lower than on 30 V nanotubes and Ti_C surfaces. The vinculin staining intensity per cell was the highest on sample 30 V with nanotubes of the largest diameter and was lower on 20 V and 10 V samples (Figure 7B). Therefore, the intensity of talin and vinculin fluorescence was higher on nanotubes with a larger diameter and larger wall thickness than on the nanotubes with smaller diameters and wall thickness.


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)

Human Saos-2 osteoblasts on 10 V, 20 V, and 30 V nanotubes, on control Ti_C, and on glass coverslips on day 3.Notes: Immunofluorescence intensity (A, B) and absorbance measured by enzyme-linked immunosorbent assay (C, D) of talin (A, C) and vinculin (B, D). Data expressed as mean ± standard error of mean. P≤0.05 considered significant in comparison with samples labeled above columns.
© Copyright Policy
Related In: Results  -  Collection

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

f7-ijn-10-7145: Human Saos-2 osteoblasts on 10 V, 20 V, and 30 V nanotubes, on control Ti_C, and on glass coverslips on day 3.Notes: Immunofluorescence intensity (A, B) and absorbance measured by enzyme-linked immunosorbent assay (C, D) of talin (A, C) and vinculin (B, D). Data expressed as mean ± standard error of mean. P≤0.05 considered significant in comparison with samples labeled above columns.
Mentions: The concentration and distribution molecular markers of cell adhesion, ie, talin and vinculin, were evaluated by immunofluorescence staining and enzyme-linked immunosorbent assay (ELISA). The immunofluorescence of talin was relatively diffused, and focal adhesions were not clearly visible (Figure S1). The fluorescence intensity of talin per cell was most intense on the 30 V nanotube sample, and there was a very low signal on glass (Figure 7A). As mentioned earlier, well-developed focal adhesion plaques containing vinculin were found on all surfaces (Figure 6). They seemed to be slightly larger on smooth Ti_C surfaces than on nanotubes. On glass, focal adhesions in Saos-2 cells were smaller than on the other surfaces, and the signal of vinculin was lower than on 30 V nanotubes and Ti_C surfaces. The vinculin staining intensity per cell was the highest on sample 30 V with nanotubes of the largest diameter and was lower on 20 V and 10 V samples (Figure 7B). Therefore, the intensity of talin and vinculin fluorescence was higher on nanotubes with a larger diameter and larger wall thickness than on the nanotubes with smaller diameters and wall thickness.

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