Highly Permeable Genipin-Cross-linked Gelatin Conduits Enhance Peripheral Nerve Regeneration

Here we have evaluated peripheral nerve regeneration with a porous biodegradable nerve conduit (PGGC), which was made from genipin-cross-linked gelatin. To examine the effect of pores, nonporous genipin-cross-linked gelatin conduit (GGC) was considered as the control. Both the PGGC and the GGC were dark blue in appearance with a concentric and round lumina. The PGGC featured an outer surface with pores of variable size homogeneously traversing, and a partially fenestrated inner surface connected by an open trabecular meshwork. The GGC had a rough outer surface whereas its inner lumen was smooth. Both PGGCs and GGCs had similar hydrophilicity on condition of the same material and cross-linking degree. The porosity of PGGCs and GGCs was 90.8 ± 0.9% and 24.3 ± 2.9%, respectively. The maximum tensile force of the GGCs (0.12 ± 0.06 kN) exceeded that of the PGGCs (0.03 ± 0.01 kN), but the PGGCs had a higher swelling ratio than GGCs at 0.5, 1, 3, 6, 12, 24, 48, 60, 72, and 84 h after soaking in deionized water. Cytotoxic testing revealed the soaking solutions of both of the tube composites would not produce cytotoxicity to cocultured Schwann cells. After subcutaneous implantation on the dorsal side of the rat, the PGGC was degraded completely after 12 weeks of implantation whereas a thin tissue capsule was formed encapsulating the partially degraded GGC. Biodegradability of both of the tube groups and their effectiveness as a guidance channel were examined as they were used to repair a 10 mm gap in the rat sciatic nerve. As a result, fragmentation of the GGC was still seen after 12 weeks of implantation, yet the PGGC had been completely degraded. Histological observation showed that numerous myelinated axons had crossed over the gap region in the PGGCs after 8 weeks of implantation despite only few myelinated axons and unmyelinated axons mostly surrounded by Schwann cells seen in the GGCs. In addition, the regenerated nerves in the PGGCs presented a significantly higher nerve conductive velocity than those in the GGCs (P < 0.05). Thus, the PGGCs can not only offer effective aids for regenerating nerves but also accelerate favorable nerve functional recovery compared with the GGCs.