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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

Scanning electron microscopy (SEM) micrographs and 2D fast fourier transform (FTT) analysis of nano-fibers collected using rotating mandrel rates of 300 (A), 1200 (B) and 2400 (C) rpm. Scale bars: 10 µm.
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ijms-16-12925-f001: Scanning electron microscopy (SEM) micrographs and 2D fast fourier transform (FTT) analysis of nano-fibers collected using rotating mandrel rates of 300 (A), 1200 (B) and 2400 (C) rpm. Scale bars: 10 µm.

Mentions: Randomly oriented fibers with an average fiber dimension of 300 nm were fabricated as previously described [28]. Aligned GL/PEO_GPTMS (gelatin/polyethylene-oxide/(3-Glycidoxypropyl)methyldiethoxysilane) nano-fibers were made using a rotating mandrel collector, and its rotation was varied from 0 to 2400 rpm to analyze the influence of this parameter on fiber alignment. FFT (2D Fast Fourier Transform) analysis of the SEM (Scanning Electron Microscopy) images was used to quantitatively analyze the degree of the GL based nano-fibers alignment. In FFT analysis, a graphical plot of frequency distribution was generated by summing the pixel intensities along the radius of the FFT output image obtained from the original SEM image. For a rotation of 2400 rpm, two sharp peaks were observed at a distance of ~180°, confirming the morphological data that showed a large number of fibers aligned in a preferential direction (Figure 1C). On the other hand, rotating at ~300 rpm produced no fiber orientation, confirmed by SEM image and FTT analysis (Figure 1A). Nanofibers size was measured at different rotating mandrel speeds showing a slight reduction in the fiber average diameters when the speed is increased. The measured diameters were 204 ± 48 nm for aligned nanofibers using a rotating speed of 2400 rpm, 238.9 ± 74 nm for 1200 rpm and 254.7 ± 68.5 nm for 300 rpm. No significant differences in fiber size were observed from fibers collected on plane or rotating collector. Nanofibers obtained using a rotating speed of 2400 rpm were used for cell test, as they showed a high degree of alignment on a preferential direction (as confirmed by FFT and SEM analysis)


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)

Scanning electron microscopy (SEM) micrographs and 2D fast fourier transform (FTT) analysis of nano-fibers collected using rotating mandrel rates of 300 (A), 1200 (B) and 2400 (C) rpm. Scale bars: 10 µm.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-16-12925-f001: Scanning electron microscopy (SEM) micrographs and 2D fast fourier transform (FTT) analysis of nano-fibers collected using rotating mandrel rates of 300 (A), 1200 (B) and 2400 (C) rpm. Scale bars: 10 µm.
Mentions: Randomly oriented fibers with an average fiber dimension of 300 nm were fabricated as previously described [28]. Aligned GL/PEO_GPTMS (gelatin/polyethylene-oxide/(3-Glycidoxypropyl)methyldiethoxysilane) nano-fibers were made using a rotating mandrel collector, and its rotation was varied from 0 to 2400 rpm to analyze the influence of this parameter on fiber alignment. FFT (2D Fast Fourier Transform) analysis of the SEM (Scanning Electron Microscopy) images was used to quantitatively analyze the degree of the GL based nano-fibers alignment. In FFT analysis, a graphical plot of frequency distribution was generated by summing the pixel intensities along the radius of the FFT output image obtained from the original SEM image. For a rotation of 2400 rpm, two sharp peaks were observed at a distance of ~180°, confirming the morphological data that showed a large number of fibers aligned in a preferential direction (Figure 1C). On the other hand, rotating at ~300 rpm produced no fiber orientation, confirmed by SEM image and FTT analysis (Figure 1A). Nanofibers size was measured at different rotating mandrel speeds showing a slight reduction in the fiber average diameters when the speed is increased. The measured diameters were 204 ± 48 nm for aligned nanofibers using a rotating speed of 2400 rpm, 238.9 ± 74 nm for 1200 rpm and 254.7 ± 68.5 nm for 300 rpm. No significant differences in fiber size were observed from fibers collected on plane or rotating collector. Nanofibers obtained using a rotating speed of 2400 rpm were used for cell test, as they showed a high degree of alignment on a preferential direction (as confirmed by FFT and SEM analysis)

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