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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

Proliferation assay: Fluorescent images (20× magnification) after DAPI (blue) and phalloidin (red) staining of RT4-D6P2T (A) and primary SC (B) on poly-l-lysine coated coverslips (control condition), random fibers and aligned fibers after 1, 3, 5, and 7 DIV (days in vitro) after seeding. Scale bar: 100 μm; RT4-D6P2T (C) and primary SC (D) cell number is expressed as cells/mm2 ± standard error of the mean (SEM). Asterisks refer to significant statistical difference with **p ≤ 0.01 and ***p ≤ 0.001.
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ijms-16-12925-f003: Proliferation assay: Fluorescent images (20× magnification) after DAPI (blue) and phalloidin (red) staining of RT4-D6P2T (A) and primary SC (B) on poly-l-lysine coated coverslips (control condition), random fibers and aligned fibers after 1, 3, 5, and 7 DIV (days in vitro) after seeding. Scale bar: 100 μm; RT4-D6P2T (C) and primary SC (D) cell number is expressed as cells/mm2 ± standard error of the mean (SEM). Asterisks refer to significant statistical difference with **p ≤ 0.01 and ***p ≤ 0.001.

Mentions: After three (p < 0.001), five (p < 0.001) and seven (p < 0.001) days, RT4-D6P2T cells proliferated more slowly when seeded on aligned fibers than under control conditions or random fibers (Figure 3C). After five (p < 0.01) and seven (p < 0.01) days, primary SC also had slower proliferation rates on aligned fibers than under control conditions or random fibers (Figure 3D). Both RT4-D6P2T and primary SC cells reached confluence under all conditions tested.


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)

Proliferation assay: Fluorescent images (20× magnification) after DAPI (blue) and phalloidin (red) staining of RT4-D6P2T (A) and primary SC (B) on poly-l-lysine coated coverslips (control condition), random fibers and aligned fibers after 1, 3, 5, and 7 DIV (days in vitro) after seeding. Scale bar: 100 μm; RT4-D6P2T (C) and primary SC (D) cell number is expressed as cells/mm2 ± standard error of the mean (SEM). Asterisks refer to significant statistical difference with **p ≤ 0.01 and ***p ≤ 0.001.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-16-12925-f003: Proliferation assay: Fluorescent images (20× magnification) after DAPI (blue) and phalloidin (red) staining of RT4-D6P2T (A) and primary SC (B) on poly-l-lysine coated coverslips (control condition), random fibers and aligned fibers after 1, 3, 5, and 7 DIV (days in vitro) after seeding. Scale bar: 100 μm; RT4-D6P2T (C) and primary SC (D) cell number is expressed as cells/mm2 ± standard error of the mean (SEM). Asterisks refer to significant statistical difference with **p ≤ 0.01 and ***p ≤ 0.001.
Mentions: After three (p < 0.001), five (p < 0.001) and seven (p < 0.001) days, RT4-D6P2T cells proliferated more slowly when seeded on aligned fibers than under control conditions or random fibers (Figure 3C). After five (p < 0.01) and seven (p < 0.01) days, primary SC also had slower proliferation rates on aligned fibers than under control conditions or random fibers (Figure 3D). Both RT4-D6P2T and primary SC cells reached confluence under all conditions tested.

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