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Development and evaluation of artificial nerve which could enhance axonal elongation in peripheral and central nervous system

增强周围和中枢神经系统轴突伸长的人工神经的研制和评价

基本信息

批准号：
11307037
负责人：
NISHIMURA Yoshihiko
金额：
$ 23.5万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11307037/
关键词：
neural stem cell spinal cord regeneration central system injury immunohistochemistry sciatic nerve peripheral nervous system アルギン酸神経再生脊髄再生末梢神経再生

项目摘要

Alginate, derived from brown seaweeds was chemically crosslinked with covalent bond. Biodegradation speed can be controlled by initial concentration of alginate and dose of irradiation. For peripheral nerve repair, sciatic nerve gap in the cats was bridged using alginate sponge, which was specially designed to biodegradate within 3 months. Three months after operation, many regenerated axon were found in the middle of the gap. Electrophysiologically, muscle action potentials could be recorded at the gastrocnemius muscle after electrical stimulation of the sciatic nerve proximal to the gap. Rat sciatic nerve gap model, regenerating axons can be seen in the biodegrading alginate, which does not have sponge structure. For central nerve repair, freeze-dried alginate sponge was implanted into the gap of rat spinal cord. Regenerating axons was seen in the implanted alginate and in the host spinal cord, which was examined by HRP-tracing study. Electrophysiologically, action potential was recorded beyond the gap. Next, hippocampus-derived neurospheres were prepared from transgenic rat fetuses expressing green fluorescent protein (GFP), and transplanted into an alginate-filled lesion of young rat spinal cord. One, two and four weeks after transplantation, a large number of grafted cells survived, many of which expressed immunoreactiveity for glial fibrillary acidic protein, and a few expressed immunoreactiveity for beta-tubulin These findings demonstrate that rat fetal hippocampus-derived neurosphere cells could survive, differentiate, and integrate well into the host spinal cord tissue.

以褐藻为原料，通过共价键对海藻酸钠进行化学交联。藻酸盐的初始浓度和辐照剂量可以控制藻酸盐的降解速度。对于周围神经修复，使用藻酸盐海绵桥接猫的坐骨神经间隙，藻酸盐海绵被专门设计为在3个月内生物降解。术后3个月，在差距中间发现许多再生轴突。电生理上，电刺激差距附近的坐骨神经后，可在腓肠肌记录到肌肉动作电位。大鼠坐骨神经缺损模型中，可生物降解的海藻酸钠中可见再生轴突，其不具有海绵状结构。将冷冻干燥的海藻酸钠海绵植入大鼠脊髓差距内进行中枢神经修复。HRP示踪研究显示，移植藻酸盐的脊髓和宿主脊髓中均可见再生轴突。在电生理学上，记录超过差距的动作电位。接着，从表达绿色荧光蛋白（GFP）的转基因大鼠胎儿制备海马衍生的神经球，并将其移植到年轻大鼠脊髓的藻酸盐填充病变中。移植后1、2、4周，移植细胞大量存活，其中多数表达胶质细胞酸性蛋白，少数表达β-微管蛋白。这些结果表明，大鼠胚胎海马神经球细胞能存活、分化并与宿主脊髓组织融合。