Fukushima Kazuyuki, Enomoto Mitsuhiro, Tomizawa Shoji, Takahashi Makoto, Wakabayashi Yoshiaki, Itoh Soichiro, Kuboki Yoshinori, Shinomiya Kenichi
Department of Orthopaedic and Spinal Surgery, Tokyo Medical and Dental University Graduate School, 1-5-45 Yushima Bunkyo-ku Tokyo, 113-8519 Japan.
J Med Dent Sci. 2008 Mar;55(1):71-9.
We developed a honeycomb-shaped lyophilized Type I atelocollagen (Honeycomb Collagen: HC) with different pore sizes, and the effectiveness of the honeycomb shape on nerve regeneration was examined. We analyzed neurite outgrowth of dorsal root ganglion (DRG) explants on HC, both in vitro and, with direct implantation of HC into the defects of adult rat spinal cords, in vivo. The neurites of DRGs on HC extended linearly through the pores. HC with a 400 microm-pore size enhanced neurite extension, and YIGSR laminin peptide coating to the HC extended more neurites than fibronectin coating. The HC scaffolds coated with YIGSR were implanted into 2 mm-defects of spinal cords at the level of T8-9. Four weeks after implantation, the implants had degraded and been replaced with self-tissues, repairing the injured site. Neurofilament-positive fibers were observed in the implantation area and passed the borders between the HC and spinal cord stumps. Functionally, a motor-evoked potential was observed in the quadriceps femoris muscle 10 weeks after implantation. The electrophysiological examination showed reconstruction of axon tracts over the implant. This result indicates that our developed honeycomb shape is advantageous for host spinal cord compared to the random pored sponge shape, and that it promotes axonal regeneration after spinal cord injury.
我们研发了具有不同孔径的蜂窝状冻干I型去端胶原蛋白(蜂窝状胶原蛋白:HC),并研究了蜂窝形状对神经再生的有效性。我们在体外分析了背根神经节(DRG)外植体在HC上的神经突生长情况,并且在体内将HC直接植入成年大鼠脊髓损伤处进行了研究。DRG在HC上的神经突通过孔隙呈线性延伸。孔径为400微米的HC促进了神经突的延伸,并且用YIGSR层粘连蛋白肽包被的HC比用纤连蛋白包被的HC延伸出更多的神经突。将包被有YIGSR的HC支架植入T8 - 9水平的2毫米脊髓损伤处。植入四周后,植入物发生降解并被自身组织替代,修复了损伤部位。在植入区域观察到神经丝阳性纤维,并穿过HC与脊髓残端之间的边界。在功能方面,植入10周后在股四头肌中观察到运动诱发电位。电生理检查显示植入物上方的轴突束得到重建。这一结果表明,与随机孔隙的海绵形状相比,我们研发的蜂窝形状对宿主脊髓更有利,并且它能促进脊髓损伤后的轴突再生。