Limits...
The Effect of Electrospun Gelatin Fibers Alignment on Schwann Cell and Axon Behavior and Organization in the Perspective of Artificial Nerve Design.

Gnavi S, Fornasari BE, Tonda-Turo C, Laurano R, Zanetti M, Ciardelli G, Geuna S - Int J Mol Sci (2015)

Bottom Line: Aligned nano-fibers reduced adhesion and proliferation rate compared with random fibers.B5011 neuron-like cells were aligned and had parallel axon growth when cultured on the aligned gelatin fibers.The data show that the alignment of electrospun gelatin fibers can modulate Schwann cells and axon organization in vitro, suggesting that this substrate shows promise as an internal filler for the design of artificial nerves for peripheral nerve reconstruction.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Biological Sciences, University of Torino, Orbassano 10043, Italy. sara.gnavi@unito.it.

ABSTRACT
Electrospun fibrous substrates mimicking extracellular matrices can be prepared by electrospinning, yielding aligned fibrous matrices as internal fillers to manufacture artificial nerves. Gelatin aligned nano-fibers were prepared by electrospinning after tuning the collector rotation speed. The effect of alignment on cell adhesion and proliferation was tested in vitro using primary cultures, the Schwann cell line, RT4-D6P2T, and the sensory neuron-like cell line, 50B11. Cell adhesion and proliferation were assessed by quantifying at several time-points. Aligned nano-fibers reduced adhesion and proliferation rate compared with random fibers. Schwann cell morphology and organization were investigated by immunostaining of the cytoskeleton. Cells were elongated with their longitudinal body parallel to the aligned fibers. B5011 neuron-like cells were aligned and had parallel axon growth when cultured on the aligned gelatin fibers. The data show that the alignment of electrospun gelatin fibers can modulate Schwann cells and axon organization in vitro, suggesting that this substrate shows promise as an internal filler for the design of artificial nerves for peripheral nerve reconstruction.

No MeSH data available.


Related in: MedlinePlus

50B11 differentiation: Confocal images (40× magnification) after DAPI (blue) and β-tubulin (red) staining of B5011 seeded on poly-l-lysine coated coverslips (control condition), random fibers and aligned fibers 24 h after forskolin treatment. Scale bar: 50 μm. (A) 50B11 cell number is expressed as cells/mm2 ± standard error of the mean (SEM) (B); Neurite length is expressed in μm (C); Asterisks refer to significant statistical difference with ***p ≤ 0.001.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4490479&req=5

ijms-16-12925-f005: 50B11 differentiation: Confocal images (40× magnification) after DAPI (blue) and β-tubulin (red) staining of B5011 seeded on poly-l-lysine coated coverslips (control condition), random fibers and aligned fibers 24 h after forskolin treatment. Scale bar: 50 μm. (A) 50B11 cell number is expressed as cells/mm2 ± standard error of the mean (SEM) (B); Neurite length is expressed in μm (C); Asterisks refer to significant statistical difference with ***p ≤ 0.001.

Mentions: 50B11 were seeded under control condition, and also on aligned and random fibers. After 24 h, the adherent cells were counted and their morphology examined by β-tubulin and DAPI staining. Addition of 75 μM forskolin resulted in 50B11 differentiation under all conditions tested (Figure 5). The alignment of gelatin electrospun fibers did not affect the number of adherent 50B11 cells (Figure 5B). After forskolin, 50B11 cells stopped proliferating and started differentiating, resulting in a reduced cell number (p < 0.001) compared with non-treated conditions (Figure 5B). Immunostaining of β-tubulin showed that 50B11 cell maintain their ability to differentiate on gelatin electrospun fibers. Confocal images showed that aligned fibers made neurites align in parallel to the direction of the fibers. Neurites growth on aligned fibers was similar to random fibers and control condition (Figure 5A). There were no differences in 50B11 neurites length under all conditions tested (Figure 5C).


The Effect of Electrospun Gelatin Fibers Alignment on Schwann Cell and Axon Behavior and Organization in the Perspective of Artificial Nerve Design.

Gnavi S, Fornasari BE, Tonda-Turo C, Laurano R, Zanetti M, Ciardelli G, Geuna S - Int J Mol Sci (2015)

50B11 differentiation: Confocal images (40× magnification) after DAPI (blue) and β-tubulin (red) staining of B5011 seeded on poly-l-lysine coated coverslips (control condition), random fibers and aligned fibers 24 h after forskolin treatment. Scale bar: 50 μm. (A) 50B11 cell number is expressed as cells/mm2 ± standard error of the mean (SEM) (B); Neurite length is expressed in μm (C); Asterisks refer to significant statistical difference with ***p ≤ 0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490479&req=5

ijms-16-12925-f005: 50B11 differentiation: Confocal images (40× magnification) after DAPI (blue) and β-tubulin (red) staining of B5011 seeded on poly-l-lysine coated coverslips (control condition), random fibers and aligned fibers 24 h after forskolin treatment. Scale bar: 50 μm. (A) 50B11 cell number is expressed as cells/mm2 ± standard error of the mean (SEM) (B); Neurite length is expressed in μm (C); Asterisks refer to significant statistical difference with ***p ≤ 0.001.
Mentions: 50B11 were seeded under control condition, and also on aligned and random fibers. After 24 h, the adherent cells were counted and their morphology examined by β-tubulin and DAPI staining. Addition of 75 μM forskolin resulted in 50B11 differentiation under all conditions tested (Figure 5). The alignment of gelatin electrospun fibers did not affect the number of adherent 50B11 cells (Figure 5B). After forskolin, 50B11 cells stopped proliferating and started differentiating, resulting in a reduced cell number (p < 0.001) compared with non-treated conditions (Figure 5B). Immunostaining of β-tubulin showed that 50B11 cell maintain their ability to differentiate on gelatin electrospun fibers. Confocal images showed that aligned fibers made neurites align in parallel to the direction of the fibers. Neurites growth on aligned fibers was similar to random fibers and control condition (Figure 5A). There were no differences in 50B11 neurites length under all conditions tested (Figure 5C).

Bottom Line: Aligned nano-fibers reduced adhesion and proliferation rate compared with random fibers.B5011 neuron-like cells were aligned and had parallel axon growth when cultured on the aligned gelatin fibers.The data show that the alignment of electrospun gelatin fibers can modulate Schwann cells and axon organization in vitro, suggesting that this substrate shows promise as an internal filler for the design of artificial nerves for peripheral nerve reconstruction.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Biological Sciences, University of Torino, Orbassano 10043, Italy. sara.gnavi@unito.it.

ABSTRACT
Electrospun fibrous substrates mimicking extracellular matrices can be prepared by electrospinning, yielding aligned fibrous matrices as internal fillers to manufacture artificial nerves. Gelatin aligned nano-fibers were prepared by electrospinning after tuning the collector rotation speed. The effect of alignment on cell adhesion and proliferation was tested in vitro using primary cultures, the Schwann cell line, RT4-D6P2T, and the sensory neuron-like cell line, 50B11. Cell adhesion and proliferation were assessed by quantifying at several time-points. Aligned nano-fibers reduced adhesion and proliferation rate compared with random fibers. Schwann cell morphology and organization were investigated by immunostaining of the cytoskeleton. Cells were elongated with their longitudinal body parallel to the aligned fibers. B5011 neuron-like cells were aligned and had parallel axon growth when cultured on the aligned gelatin fibers. The data show that the alignment of electrospun gelatin fibers can modulate Schwann cells and axon organization in vitro, suggesting that this substrate shows promise as an internal filler for the design of artificial nerves for peripheral nerve reconstruction.

No MeSH data available.


Related in: MedlinePlus