Abstract:
Background Tissue engineered peripheral nerve regeneration materials focus on restoring the peripheral nerve regeneration microenvironment. Traditional polycaprolactone-serine electrostatic spinning materials can mimic the topological cues of the peripheral nerve matrix, but lack of regeneration-related biological sites.
Objective To combine the Schwann cells extracellular matrix with electrostatic spinning materials thus reproducing the peripheral nerve regeneration microenvironment in both physical and bioinformatic terms, and investigate the effect of this composite material on nerve fiber regeneration as well as on the Schwann cells.
Methods Rat primary Schwann cells were extracted, purified and proliferated in culture. Polycaprolactone was co-spun with silk to obtain an oriented electrospun material, purified Schwann cells were grown on the material and extracellular matrix secretion was stimulated using ascorbic acid, decellularization was performed using Triton X-100 and ammonia to obtain a cell-free composite. The material was characterized by scanning electron microscopy as well as by immunofluorescence chemical staining. After verification of the successful assembly of the composite, rat dorsal root ganglion tissue was grown on the surface of the composite and the purely oriented material was used as a comparison; after 7 days of dorsal root ganglion growth, immunofluorescence staining was performed to assess the axonal growth rate as well as the migration ability of Schwann cells.
Results The cells were successfully extracted, purified and proliferated. Electron microscopic and immunofluorescence results demonstrated that the extracellular matrix of Schwann cells was successfully compounded with electrospun nanofibers and that the oriented fiber morphology was retained, only a small amount of DNA remained after decellularization (2.398 ± 0.233 ng/mg). The extension length of dorsal root ganglion axons on the composite surface was significantly greater than that of the scaffold material alone (1503 ± 147.4 μm vs 567.7 ± 34.63 μm, P＜0.000 1). The migration distance of Schwann cells in the composite was greater than the extension of axons (2073 ± 112.9 μm vs 1503 ± 147.4 μm, P=0.005 6).
Conclusion The strategy of combining the extracellular matrix of Schwann cells with electrospinning materials can support the rapid growth of DRG axons and the rapid migration of Schwann cells.
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