Efficacy of Freeze-thawed Peripheral Nerve Allograft in the Dog

冻融狗同种异体周围神经移植物的疗效

• 批准号: 12833002
• 负责人: TOMIZAWA Nobuyuki
• 金额: $ 2.18万
• 依托单位: Iwate 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 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12833002/
• 关键词: freeze-thawed nerve allograft; peripheral nerve; Schwann cell; dog; rat; 同種冷凍神経移植

To obtain the fundamental information firstly, the functional recovery from peripheral nerve defects was studied by carrying out freeze-thawed nerve allograft and autograft of rat sciatic nerves. Although several neurological examinations were performed after nerve grafting, only superficial pain perception was recovered within the 110 days postoperatively in most animals. Sciatic function index was not effective to evaluate the motor nerve function because of individual deviations.To investigate the efficacy of Schwann cell (SC) transplantation with freeze-thawed peripheral nerve allograft, 4 types of SC cultures from adult rat sciatic nerves were compared. The method by Calderon-Martinez was the most excellent technique on viability and purity of SCs. The same method for culturing SCs was performed in adult beagle dogs. As a result, the purity of SCs was about 90% and the number of SCs was about 3.6×10^7 from 3cm of dog sciatic nerve.In order to evaluate the application of SC cultures along with freeze-thawed nerve allograft, co-culture of SCs and freeze-thawed nerve grafts, and the direct injection of cultured SCs to freeze-thawed nerve grafts were performed. No living SCs were observed in co-cultured nerve graft. On the other hand, there were many viable SCs in nerve graft by the direct injection of cultured SCs to freeze-thawed nerve grafts.From the results of this study, it was elucidated that freeze-thawed nerve allograft was effective on the recovery of sensory function, and SC culture from adult rat sciatic nerves was available for promotion of nerve regeneration. Moreover, it was proved that SC culture from adult dogs was also possible, and the direct injection of cultured SCs to nerve grafts was effective for the application of SC cultures along with freeze-thawed nerve allograft.

为了首先获得基本信息，本研究采用冻融同种异体神经移植和大鼠自体坐骨神经移植的方法，对周围神经损伤后的功能恢复进行了研究。虽然神经移植后进行了几次神经学检查，但大多数动物在术后110天内只恢复了表面的痛觉。由于个体差异，坐骨神经功能指数不能有效地评价运动神经功能。为探讨雪旺细胞(SC)与冻融同种异体周围神经移植的效果，比较了4种培养的成年大鼠坐骨神经SC。Calderon-Martinez的方法在干细胞活性和纯度方面是最好的方法。同样的方法在成年比格犬身上进行干细胞的培养。结果表明，从犬坐骨神经3 cm处获得的SCs纯度约为90%，细胞数约为3.6×107。为评价SC培养与冻融同种异体神经移植的应用，将培养的SCs与冻融神经移植物共培养，并将培养的SCs直接注入冻融神经移植物中。在共培养的神经移植物中没有观察到存活的干细胞。另一方面，通过将培养的SCs直接注入冻融神经移植物中，移植的神经中有大量的SCs存活，说明冻融同种异体神经移植对感觉功能的恢复是有效的，成年大鼠坐骨神经的SC培养可以促进神经再生。此外，成年犬的SC培养也是可行的，将培养的SC直接注射到神经移植物中是与冻融同种异体神经移植一起应用的有效方法。