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

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.

Controlling the morphology and outgrowth of nerve and neuroglial cells: The effect of surface topography

“…Filaments or fibers have been made of synthetic polymers -like polypropylene (PP) [79], polycaprolactone (PCL) [80], poly-L-lactide (PLLA) [71], poly(acrylonitrile-comethylacrylate (PAN-MA) [48,70] -and natural polymers (e.g. gelatin [63] and chitosan [81]).…”

“…The effect of topography on cells of neuroglial phenotype has been studied mainly on dissociated primary Schwann cells [58 62], but there are few studies using Schwann cell lines, like RT4-D6P2T [63] and CRL-2765 [64]. Studies on the cellular response of astrocytes on micropatterned substrates remain limited [65,66].…”

“…To our knowledge, there is no additional study on the effect of oligodendrocyes onto micropatterned substrate. Interestingly, cell adhesion and proliferation were deteriorated on the aligned vs. the random fibers [63]. When Schwann cells were cultured on aligned electrospun chitosan fibers of wider diameter (i.e.…”

“…Schwann cells on parallel aligned fibers of subcellular size change the shape and orientation state they exhibit on the smooth films and become oriented and aligned along the fibers axis [59,62,63,81].…”

Peptide nanostructures on nanofibers for peripheral nerve regeneration

“…Our results are comparable with other literature reports where the nanofibrous scaffolds showed lower proliferation rates of Human embryo skin fibroblasts (Duan et al, ) and neural stem cells (Christopherson, Song, & Mao, ) than the TCPS control. Gnavi et al () reported that aligned nanofibers have shown that SCs (RT4‐D6P2T) proliferated more slowly than under TCPS control conditions. It may be attributed to the fact that nanofibrous scaffolds promote better cell spreading and extension compared with the TCPS control, which reduces the space available for the new cell adhesion and thus lower proliferation rates.…”