Effective integration of stimulation and direction in bionic scaffolds by materials and microstructure design has been the focus in the advancement of nerve regeneration. Hydrogels are the most promising biomimicked materials used in developing nerve grafts, but the highly hydrated networks limit the fabrication of hydrogel materials into complex biomedical devices. Herein, facile lithography-free and spontaneously micropatterned techniques were used to fabricate a smart protein hydrogel-based scaffold, which carried topographical, electrical, and chemical induction for neural regulation. The synthesized tissue-mimicked silk-gelatin (SG)/polylactic acid bilayer system can self-form three-dimensional ordered corrugation micropatterns with well-defined dimensions (wavelength, λ) based on the stress-induced topography. Through magnetically and topographically guided deposition of the synthesized nerve growth factor-incorporated Fe3O4-graphene nanoparticles (GFPNs), a biologically and electrically conductive cell passage with one-dimensional directionality was constructed to allow for a controllable constrained geometric effect on neuronal adhesion, differentiation, and neurite orientation. Particularly, the SG with corrugation patterns of λ ≈ 30 μm resulted in the optimal cell adhesion and differentiation in response to the pattern guidance. Furthermore, the additional electrical stimulation applied on GFPN-deposited SG resulted in a 1.5-fold increase in the neurite elongation by day 7, finally leading to the neuronal connection by day 21. Such a hydrogel device with synergistic effects of physical and chemical enhancement on neuronal activity provides an expectable opportunity in the development of next-generation nerve conduits.
Keywords: cell alignment; electrical stimulation; hydrogels; lithography-free fabrication; nerve tissue scaffold; silk.