Development of the Artificial Nerve Using Laminin Peptides and rhMDP

我们开发了具有共价结合多肽特性的肌腱壳聚糖管，并在体内研究了层粘连蛋白肽偶联于这些管壁对神经再生的有效性。采用三角形截面的壳聚糖管将15mm的桥移植物植入SD大鼠坐骨神经，壳聚糖管含有共价结合的完整层粘连蛋白、层粘连蛋白肽CDPGYIGSR或CSRARKQAASIKVAVSAD，或不处理（每组N=20）。作为对照，进行等接枝（N=5）。各组分别于1、2、4、6、8周后处死3只进行组织学观察。测量再生组织在试管内的总面积和再生组织附着在试管内表面的面积长度。实验组和对照组各取5只大鼠，在植入12周后测定植入部位与未治疗部位之间的潜伏期商。此外，在远端吻合部位10mm处测量有髓鞘轴突面积的百分比。组织学结果表明，术后12周免疫染色证实的固定层粘连蛋白以及层粘连蛋白寡肽可以有效地辅助神经组织伸展。根据与诱发动作电位相关的神经组织百分比的统计分析，首先使用YIGSR然后使用IKVAV的顺序治疗在增强神经再生方面的效果与完整层粘连蛋白相匹配。然而，与等接相比，再生轴突生长的增强不够充分。

We have developed tendon chitosan tubes having the property to covalently bind peptides, and the effectiveness of laminin peptides coupled to these tubular wall on nerve regeneration was examined in vivo. Bridge graft implantation (15mm) into the sciatic nerve of SD rats was carried out using chitosan tubes having a triangular cross section containing either covalently bound intact laminin, the laminin peptides CDPGYIGSR or CSRARKQAASIKVAVSAD, or being non-treated (N=20 in each group). As a control, isografting (N=5) was carried out. Three rats in each experimental group were sacrificed for histology observations after respectively 1, 2, 4, 6 and 8 weeks. The total area of regenerating tissue in the tube and the length of the area where regenerating tissue attached to the inner surface of the tube were measured. In five rats from each experimental and control group, the latency quotient between the implanted and non-treated site was determined 12 weeks after implantation. Furthermore, the percentage of the myelinated axon area was measured at a 10 mm distance of the distal anastomosed site. Histological findings suggest that the immobilized laminin confirmed by immunostain as long as 12 weeks post operatively, as well as laminin oligopeptides may effectively assist nerve tissue extension. According to statistical analysis of the percentage neural tissue found in relation to evoked action potentials, the sequential treatments with YIGSR first followed by IKVAV matched the effectiveness of intact laminin in order to enhance nerve regeneration. However, when compared with isografting, the enhancement of regenerated axon growth was less sufficient.