Kaizawa Yukitoshi, Kakinoki Ryosuke, Ikeguchi Ryosuke, Ohta Soichi, Noguchi Takashi, Takeuchi Hisataka, Oda Hiroki, Yurie Hirofumi, Matsuda Shuichi
Cell Transplant. 2017 Feb 16;26(2):215-228. doi: 10.3727/096368916X692951. Epub 2016 Sep 21.
Cells, scaffolds, growth factors, and vascularity are essential for nerve regeneration. Previously, we reported that the insertion of a vascular bundle and the implantation of bone marrow-derived mesenchymal stem cells (BM-MSCs) into a nerve conduit promoted peripheral nerve regeneration. In this study, the efficacy of nerve conduits containing a vascular bundle, BM-MSCs, and thermally decellularized allogenic nerve matrix (DANM) was investigated using a rat sciatic nerve model with a 20-mm defect. Lewis rats were used as the sciatic nerve model and for the preparation of BM-MSCs, and Dark Agouti rats were used for the preparation of the DANM. The revascularization and the immunogenicity of the DANM were investigated histologically. The regeneration of nerves through nerve conduits containing vessels, BM-MSCs, and DANM (VBD group) was evaluated based on electrophysiological, morphometric, and reinnervated muscle weight measurements and compared with that of vessel-containing conduits that were implanted with BM-MSCs (VB group). The DANM that was implanted into vessel-containing tubes (VCTs) was revascularized by neovascular vessels that originated from the inserted vascular bundle 5-7 days after surgery. The number of CD8+ cells found in the DANM in the VCT was significantly smaller than that detected in the untreated allogenic nerve segment. The regenerated nerve in the VBD group was significantly superior to that in the VB group with regard to the amplitude of the compound muscle action potential detected in the pedal adductor muscle; the number, diameter, and myelin thickness of the myelinated axons; and the tibialis anterior muscle weight at 12 and 24 weeks. The additional implantation of the DANM into the BM-MSC-implanted VCT optimized the axonal regeneration through the conduit. Nerve conduits constructed with vascularity, cells, and scaffolds could be an effective strategy for the treatment of peripheral nerve injuries with significant segmental defects.
细胞、支架、生长因子和血管形成对于神经再生至关重要。此前，我们报道将血管束插入并将骨髓间充质干细胞（BM-MSCs）植入神经导管可促进周围神经再生。在本研究中，使用大鼠坐骨神经20毫米缺损模型，研究了含有血管束、BM-MSCs和热脱细胞同种异体神经基质（DANM）的神经导管的疗效。Lewis大鼠用作坐骨神经模型并用于制备BM-MSCs，Dark Agouti大鼠用于制备DANM。对DANM的血管再生和免疫原性进行了组织学研究。基于电生理、形态测量和再支配肌肉重量测量，评估通过含有血管、BM-MSCs和DANM的神经导管（VBD组）的神经再生情况，并与植入BM-MSCs的含血管导管（VB组）进行比较。植入含血管管（VCTs）的DANM在术后5-7天被源自插入血管束的新生血管重新血管化。在VCT中DANM中发现的CD8+细胞数量明显少于未处理的同种异体神经段中检测到的数量。在踏板内收肌中检测到的复合肌肉动作电位的幅度、有髓轴突的数量、直径和髓鞘厚度以及12周和24周时的胫前肌重量方面，VBD组的再生神经明显优于VB组。将DANM额外植入植入BM-MSCs的VCT可优化通过导管的轴突再生。构建具有血管、细胞和支架的神经导管可能是治疗具有明显节段性缺损的周围神经损伤的有效策略。
