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，并将层粘连蛋白多肽偶联到这些管壁上，在体内检测了其对神经再生的效果。用含有共价结合的完整层粘连蛋白、层粘连蛋白多肽CDPGYIGSR或CSRARKQAASIKVAVSAD的三角形壳聚糖管进行SD大鼠坐骨神经桥接移植(15 Mm)，或不处理的壳聚糖管(每组20只)。作为对照，进行同种异体移植(N=5)。实验1、2、4、6、8周后，每组各处死3只大鼠进行组织学观察。测量管内再生组织的总面积和再生组织附着在管内表面的区域长度。每组大鼠各5只，分别于植入后12周测定植入部位与未处理部位的潜伏商。此外，在远端吻合处10 mm处测量有髓轴突面积的百分比。组织学结果提示，术后12周免疫组织化学证实固定化的层粘连蛋白以及层粘连蛋白寡肽可能有效地促进神经组织的伸展。根据与诱发动作电位相关的神经组织百分比的统计分析，先使用YIGSR，然后使用IKVAV的顺序治疗与完整的层粘连蛋白相匹配，以促进神经再生。然而，与同种异体移植相比，促进再生轴突生长的作用还不够充分。

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.