Limits...
Glycine-spacers influence functional motifs exposure and self-assembling propensity of functionalized substrates tailored for neural stem cell cultures.

Taraballi F, Natalello A, Campione M, Villa O, Doglia SM, Paleari A, Gelain F - Front Neuroeng (2010)

Bottom Line: The understanding of phenomena involved in the self-assembling of bio-inspired biomaterials acting as three-dimensional scaffolds for regenerative medicine applications is a necessary step to develop effective therapies in neural tissue engineering.Accordingly, the longer spacer of glycines, the more effective is the functional motif in both eliciting NSCs adhesion, improving their viability and increasing their differentiation.Therefore, optimized designing strategies of functionalized biomaterials may open, in the near future, new therapies in tissue engineering and regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: Center for Nanomedicine and Tissue Engineering, A.O. Ospedale Niguarda Ca' Granda Milan, Italy.

ABSTRACT
The understanding of phenomena involved in the self-assembling of bio-inspired biomaterials acting as three-dimensional scaffolds for regenerative medicine applications is a necessary step to develop effective therapies in neural tissue engineering. We investigated the self-assembled nanostructures of functionalized peptides featuring four, two or no glycine-spacers between the self-assembly sequence RADA16-I and the functional biological motif PFSSTKT. The effectiveness of their biological functionalization was assessed via in vitro experiments with neural stem cells (NSCs) and their molecular assembly was elucidated via atomic force microscopy, Raman and Fourier Transform Infrared spectroscopy. We demonstrated that glycine-spacers play a crucial role in the scaffold stability and in the exposure of the functional motifs. In particular, a glycine-spacer of four residues leads to a more stable nanostructure and to an improved exposure of the functional motif. Accordingly, the longer spacer of glycines, the more effective is the functional motif in both eliciting NSCs adhesion, improving their viability and increasing their differentiation. Therefore, optimized designing strategies of functionalized biomaterials may open, in the near future, new therapies in tissue engineering and regenerative medicine.

No MeSH data available.


Related in: MedlinePlus

Quantitative NSCs differentiation assay. Data are expressed as percentages of cells positive for GFAP, β-Tubulin, GALC/O4 and Nestin markers over the total number of cells counted in the same fields (see Materials and Methods for details). NSCs have been differentiated for 7 days in vitro. By increasing the length of the Gly-spacer, the percentage of Nestin+ significantly decreased. Despite that GFAP+ cell fraction increased inversely. β-Tubulin+ and GalC/O4+ cell percentages did not significantly differ among the tested substrates and assembled SAPs. NSCs cultured over plastic well surfaces (Negative Control), being composed of clusters of mainly Nestin+ cells, were not included in this graph. Values are expressed as means ± standard error of the mean. * = significantly different values (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2821182&req=5

Figure 7: Quantitative NSCs differentiation assay. Data are expressed as percentages of cells positive for GFAP, β-Tubulin, GALC/O4 and Nestin markers over the total number of cells counted in the same fields (see Materials and Methods for details). NSCs have been differentiated for 7 days in vitro. By increasing the length of the Gly-spacer, the percentage of Nestin+ significantly decreased. Despite that GFAP+ cell fraction increased inversely. β-Tubulin+ and GalC/O4+ cell percentages did not significantly differ among the tested substrates and assembled SAPs. NSCs cultured over plastic well surfaces (Negative Control), being composed of clusters of mainly Nestin+ cells, were not included in this graph. Values are expressed as means ± standard error of the mean. * = significantly different values (P < 0.05).

Mentions: The same in vitro protocol was adopted to test the degree of differentiation of NSC vs Gly-spacer length at 7 days in vitro (Figure 7). Fractions of GFAP+ cells increased with spacer length and, inversely, Nestin+ cells decreased. In case of both markers, NSCs differentiated over all SAPs show values significantly different from positive control. Nonetheless percentages of β-Tubulin+ and GalC/O4+ cells did not correlate with the length of the spacers.


Glycine-spacers influence functional motifs exposure and self-assembling propensity of functionalized substrates tailored for neural stem cell cultures.

Taraballi F, Natalello A, Campione M, Villa O, Doglia SM, Paleari A, Gelain F - Front Neuroeng (2010)

Quantitative NSCs differentiation assay. Data are expressed as percentages of cells positive for GFAP, β-Tubulin, GALC/O4 and Nestin markers over the total number of cells counted in the same fields (see Materials and Methods for details). NSCs have been differentiated for 7 days in vitro. By increasing the length of the Gly-spacer, the percentage of Nestin+ significantly decreased. Despite that GFAP+ cell fraction increased inversely. β-Tubulin+ and GalC/O4+ cell percentages did not significantly differ among the tested substrates and assembled SAPs. NSCs cultured over plastic well surfaces (Negative Control), being composed of clusters of mainly Nestin+ cells, were not included in this graph. Values are expressed as means ± standard error of the mean. * = significantly different values (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2821182&req=5

Figure 7: Quantitative NSCs differentiation assay. Data are expressed as percentages of cells positive for GFAP, β-Tubulin, GALC/O4 and Nestin markers over the total number of cells counted in the same fields (see Materials and Methods for details). NSCs have been differentiated for 7 days in vitro. By increasing the length of the Gly-spacer, the percentage of Nestin+ significantly decreased. Despite that GFAP+ cell fraction increased inversely. β-Tubulin+ and GalC/O4+ cell percentages did not significantly differ among the tested substrates and assembled SAPs. NSCs cultured over plastic well surfaces (Negative Control), being composed of clusters of mainly Nestin+ cells, were not included in this graph. Values are expressed as means ± standard error of the mean. * = significantly different values (P < 0.05).
Mentions: The same in vitro protocol was adopted to test the degree of differentiation of NSC vs Gly-spacer length at 7 days in vitro (Figure 7). Fractions of GFAP+ cells increased with spacer length and, inversely, Nestin+ cells decreased. In case of both markers, NSCs differentiated over all SAPs show values significantly different from positive control. Nonetheless percentages of β-Tubulin+ and GalC/O4+ cells did not correlate with the length of the spacers.

Bottom Line: The understanding of phenomena involved in the self-assembling of bio-inspired biomaterials acting as three-dimensional scaffolds for regenerative medicine applications is a necessary step to develop effective therapies in neural tissue engineering.Accordingly, the longer spacer of glycines, the more effective is the functional motif in both eliciting NSCs adhesion, improving their viability and increasing their differentiation.Therefore, optimized designing strategies of functionalized biomaterials may open, in the near future, new therapies in tissue engineering and regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: Center for Nanomedicine and Tissue Engineering, A.O. Ospedale Niguarda Ca' Granda Milan, Italy.

ABSTRACT
The understanding of phenomena involved in the self-assembling of bio-inspired biomaterials acting as three-dimensional scaffolds for regenerative medicine applications is a necessary step to develop effective therapies in neural tissue engineering. We investigated the self-assembled nanostructures of functionalized peptides featuring four, two or no glycine-spacers between the self-assembly sequence RADA16-I and the functional biological motif PFSSTKT. The effectiveness of their biological functionalization was assessed via in vitro experiments with neural stem cells (NSCs) and their molecular assembly was elucidated via atomic force microscopy, Raman and Fourier Transform Infrared spectroscopy. We demonstrated that glycine-spacers play a crucial role in the scaffold stability and in the exposure of the functional motifs. In particular, a glycine-spacer of four residues leads to a more stable nanostructure and to an improved exposure of the functional motif. Accordingly, the longer spacer of glycines, the more effective is the functional motif in both eliciting NSCs adhesion, improving their viability and increasing their differentiation. Therefore, optimized designing strategies of functionalized biomaterials may open, in the near future, new therapies in tissue engineering and regenerative medicine.

No MeSH data available.


Related in: MedlinePlus