Research of a novel tissue-engineered nerve graft for sciatic nerve repair in rats

Abstract

Objective To observe the effect of novel tissue engineered nerve grafts, which combined silk fibroin/collagen scaffold with a co-culture of Schwann cells (SCs) and adipose-derived stem cells (ADSCs) , to repair rat sciatic nerve defects. Methods From February, 2015 to August, 2016, the culture and purify SCs and ADSCs were isolated. And co-cultured at a ratio of 2∶1 and introduced into a silk fibroin (SF)/collagen scaffold to construct a tissue-engineered nerve conduit (TENC), which were transplanted to bridge 10 mm long sciatic nerve defects in rats. The experiment was divided randomly into 4 groups (10 rats/group): those bridged with plain SF/collagen scaffolds(Scaffold group), those bridged with TENCs (TENC group), those bridged with autografts (Autograft group) and those unoperated side (Normal group). The mechanical properties were examined using a universal testing machine(Instron 5865). Scanning electron microscopy was performed to observe the structure of the SF/collagen scaffold and the cells' growth. A series of electrophysiological examinations and morphological analyses were performed 12 weeks after surgery to evaluate the effect of the TENC on peripheral nerve regeneration. And One-way ANOVA was used to analyze the data. If the differences between groups were statistically significant, the Turkey's method was further applied for comparison. Results The plain SF/collagen scaffold showed appropriate pore size and good intercommunicating of holes. The cells were tightly attached to and partly coiled about the scaffold and exhibited either a spindle or a spherical shape. The results of the mechanical measurement revealed that the maximum and average Young’s moduli of the SF/collagen scaffold were (10.80 ± 0.30) MPa and (8.14 ± 0.20) MPa, respectively. The mechanical properties ensure that the scaffold could resist muscular contraction and maintain its shape unchanged for a considerable period of time after grafting. All rats in each group had achieved nerve defect regeneration in varying degrees. But in terms of the effect of the repaired nerve, those treated with TENC were similar to those with autologous nerve grafts but superior to those with plain SF/collagen scaffolds. Conclusion The TENC that combined silk fibroin/collagen scaffold with a co-culture of SCs and ADSCs had normal nerve-like structure, and can bridge sciatic nerve defect and promote nerve growth. Key words: Silk fibroin; Collagen; Schwann cells; Adipose-derived stem cells; Co-culture; Tissue engineered nerve