Experimental studies on tissue-engineered peripheral nerve using culturing of Schwann cell

雪旺细胞培养组织工程周围神经的实验研究

• 批准号: 12671439
• 负责人: NAKAO Yasushi
• 金额: $ 1.41万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671439/
• 关键词: Schwann cell; artificial nerve; nerve graft; Schawann細胞; HNK-1糖鎖抗原

Tissue engineering offers the potential to create replacement structures from autologous cells and biodegradable polymer conduits. Since they contain living cells, these structures have the potential to grow and induce regeneration. The objectives of this study are to create Schwann cells-seeded biodegradable nerve guide, and to assess in vivo nerve regeneration through the guides.A novel, biodegradable polymer conduit (2 mm in diameter, 0.5 mm in thickness, 25 mm in length) was composed of a polycaprolactone-polylactic acid copolymer reinforced with woven polyglycolic acid. First, Schwann cells were isolated from Wistar rat sciatic nerve and expanded in vitro by eight weeks. Second, these cells were seeded into the polymer conduit via a low-pressure injection and centrifuge technique. Two weeks after seeding, Schwann cells culturing in a porous conduit wall were observed using electron microscope. Third, the Schwann cells-seeded conduits were used to bridge 20-mm defects in the Wistar … More rat sciatic nerve. Polymer conduits lacking Schwann cells and silicone conduits were implanted into another set of animals as controls. After 8 weeks, sciatic nerve including conduit was harvested and fixed by glutaraldehyde. At the mid portion of the conduit, cross sections were cut and stained with Toluidine blue to evaluate axonal contents.Electron microscopic study prior to grafting showed numerous number of Schwann cell culturing in a porous conduit wall with 3-dimensional manner. Histological assessment with light microscope at 8 weeks demonstrated excellent regeneration with well-myelinated fibers in the bridging tissue through the Schwann cells-seeded polymer conduits. In the untreated polymer conduits, four animals showed bridging tissue but nerve regeneration was poor. There was no regeneration through the silicone conduits. These findings suggest that tissue-engineered devices, composed of biodegradable polymer materials and adherent living Schwann cells can facilitate nerve regeneration in reconstruction for a long nerve gap. Less

组织工程提供了从自体细胞和可生物降解的聚合物导管创建替代结构的可能性。由于它们含有活细胞，这些结构具有生长和诱导再生的潜力。本研究的目的是制备雪旺细胞种植的可生物降解神经导管，并评价其体内神经再生能力。首先，从Wistar大鼠坐骨神经中分离雪旺细胞，并在体外扩增8周。其次，通过低压注射和离心技术将这些细胞接种到聚合物导管中。接种后2周，电镜观察许旺细胞在多孔导管壁中的培养情况。第三，种植雪旺细胞的导管用于桥接Wistar大鼠20 mm的缺损。 ...更多信息 大鼠坐骨神经将缺乏雪旺细胞的聚合物导管和硅胶导管植入另一组动物作为对照。8周后，取坐骨神经带导管，戊二醛固定。在导管的中部切下横截面，用甲苯胺蓝染色以评估轴突含量，移植前的电子显微镜研究显示在多孔的导管壁中以三维方式培养了大量的雪旺细胞。在8周时用光学显微镜进行的组织学评估表明，通过施旺细胞接种的聚合物导管，桥接组织中具有良好的有髓纤维的良好再生。在未处理的聚合物导管中，4只动物显示桥接组织，但神经再生较差。通过硅胶导管没有再生。这些发现表明，由可生物降解的聚合物材料和粘附的活雪旺细胞组成的组织工程装置可以促进长神经缺损重建中的神经再生。少

项目成果

Harukazu Saito, et al.: "Specific regulation and localization of carbohydrate epitope on motor and sensory Schwann cells"J Jpn.Soc.Surg.Hand. 17. 399-402 (2000)

Harukazu Saito 等人：“运动和感觉雪旺细胞上碳水化合物表位的特异性调节和定位”J Jpn.Soc.Surg.Hand。

Noriko Okuyama, et al.: "The effect of total area of fascicle in cable grafting on axonal regeneration"J Jpn.Soc.Surg.Hand. 18. 192-195 (2001)

Noriko Okuyama 等人：“电缆移植中束总面积对轴突再生的影响”J Jpn.Soc.Surg.Hand。

仲尾保志: "末梢神経の最前線から"臨床整形外科. 35・13. 1479-1487 (2000)

中尾康：“从周围神经的前沿”临床骨科 35・13（2000）。

仲尾 保志: "再生医療工学の最先端"シーエムシー出版. 76-79 (2002)

Yasushi Nakao：“再生医学工程的前沿”CMC Publishing 76-79（2002）。

仲尾保志: "末梢神経移植の最前線から"臨床整形外科. 35・13. 1479-1487 (2000)

中尾靖：《来自周围神经移植的前沿》《临床骨科》35・13（2000）。