Suzuki Masumi, Itoh Soichiro, Yamaguchi Isamu, Takakuda Kazuo, Kobayashi Hisatoshi, Shinomiya Kenichi, Tanaka Junzo
Department of Plastic Surgery, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan.
J Neurosci Res. 2003 Jun 1;72(5):646-59. doi: 10.1002/jnr.10589.
We have developed tendon chitosan tubes having the ability to bind peptides covalently, and the effectiveness of laminin peptides coupled to these tubular wall on nerve regeneration was examined in vivo. Bridge graft implantation (15 mm) into the sciatic nerve of SD rats was carried out using chitosan tubes having a triangular cross section containing either covalently bound intact laminin or the laminin peptides CDPGYIGSR or CSRARKQAASIKVAVSAD or being nontreated (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 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 the nontreated site was determined 12 weeks after implantation. Furthermore, the percentage of myelinated axon area was measured at a 10-mm distance from the distal anastomosed site. Histological findings suggest that the immobilized laminin, confirmed by immunostaining as long as 12 weeks postoperatively, 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 enhancing nerve regeneration. However, when compared with that after isografting, the enhancement of regenerated axon growth was less sufficient.
我们研发出了具有共价结合肽能力的肌腱壳聚糖管,并在体内检测了与这些管壁偶联的层粘连蛋白肽对神经再生的有效性。使用具有三角形横截面的壳聚糖管进行桥接移植(15毫米)到SD大鼠的坐骨神经中,这些壳聚糖管含有共价结合的完整层粘连蛋白或层粘连蛋白肽CDPGYIGSR或CSRARKQAASIKVAVSAD,或者未进行处理(每组N = 20)。作为对照,进行了同基因移植(N = 5)。在1、2、4、6和8周后,每组处死3只大鼠进行组织学观察。测量管内再生组织的总面积以及再生组织附着在管内表面的区域长度。在每个实验组和对照组的5只大鼠中,在植入后12周测定植入部位与未处理部位之间的潜伏期商数。此外,在距远端吻合部位10毫米处测量有髓轴突面积的百分比。组织学结果表明,通过免疫染色证实术后长达12周仍固定存在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延伸。根据对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的序贯处理与完整层粘连蛋白在促进神经再生方面的有效性相当。然而,与同基因移植后的情况相比,再生轴突生长的增强程度不够充分。