利用激光技术对周围神经的倍数再生与重塑进行高选择性干预与分析的研究

Outgrowth of collateral sprouts from both afferent and motor axons is a natural process that arises during regeneration in the peripheral nervous system. It was generally acknowledged by physiologists and anatomists. In our previous study, we found that if there are enough distal endoneurial tubes into which the proximal regenerative axons can grow, the regenerative axon will sprout and maintain the most possible collaterals with an estimated maximum value of approximately 3.3, and suggest that the collateral sprouts might be a new donor nerve resource in clinical nerve repair. At the same time, we created a high selective nerve fiber incision instrument which can cut nerve within 100 micrometers (several neurons or fibers) by using laser techniques. In this project, we will study the electrophysiological characteristics of different kinds of peripheral nerve fibers in micrometer level in vivo, and try to draw the spectrum of their features to develop a new classification and identification method of peripheral nerve fibers based on the precisely analytic function. And then, we will try to clarify the distribution of all these fibers in the spinal cord and the transmission characteristics of neural signal and their integration patterns. Also, we will investigate the regulation the remodeling mechanism among different neural fibers and that with their effectors in spinal cord and brain. Thus, we will greatly improve the operating technique of peripheral into the level of micrometers, and develop new repairing methods for peripheral nerve injury to improve restoration effects.

周围神经再生过程中能够形成数倍的新生轴芽，被解剖和生理学家公认。课题组前期研究阐明了这些倍数再生轴芽的临床价值，明确了倍数再生极限为3.3，并提出了利用倍数再生机制修复严重神经损伤的可行性方案。与此同时课题组将激光技术与医学知识交叉，自主研制了微米级神经纤维切割仪，可在100微米级别进行高选择性切割（几-几十神经元/纤维）。本研究将在此基础上1、在微米级别研究不同种类神经纤维的在体电生理特性，绘制其分类特征图谱，发展基于功能精确分析的周围神经纤维新的分类与标识方法；2、阐明生理状态下周围神经各种纤维成分在脊髓层面的分布规律及神经信号传递特征与整合模式；3、深入探讨周围神经损伤修复倍数再生过程中，不同种类神经纤维间、及其与效应器和各级中枢之间的相互调节与重塑机制。从而将周围神经操作技术提高到微米级别；有依据地发展新的周围神经损伤修复方法；改善周围神经损伤的修复效果。

周围神经再生过程中能够形成数倍的新生轴芽，被解剖和生理学家公认。课题组前期研究阐明了这些倍数再生轴芽的临床价值，明确了倍数再生极限为3.3，并提出了利.用倍数再生机制修复严重神经损伤的可行性方案。本课题将激光技术与医学知识交叉，利用自主研制的微米级神经纤维切割仪，在100 微米级别进行高选择性切割（几-几十神经元/纤维），同时结合改进的多重标记方法，以及组织光透明技术，从微米尺度对周围神经进行标记、示踪、并进行结构信息的获取与解析；并对周围神经损伤倍数再生修复过程中的适应性改变做了进一步的探讨,重点如下：.1、以激光技术为基础，结合组织透明与改良标记，发展了一种在微米尺度对周围神经从肌肉-神经干-脊髓进行整体成像的新方法；.2、在微米尺度阐明了周围神经主要纤维成分在神经干中的分布以及周围神经运动成分在脊髓前角的三维分布特征及其相互关系；.3、获得肢体主要神经干及其分支神经信号传递特征与在脑中的反馈时空特性，并绘制了特征性分析图谱；.4、整体获取了神经肌肉接头在小鼠多种骨骼中的空间结构信息与三维分布特征；.5、从整体角度初步阐明了周围神经损伤倍数再生修复过程中脊髓运动神经元池重塑的结构基础。. 本研究的结果为深入理解周围神经损伤修复的结构基础提供了数据支撑，为利用倍数再生的神经纤维修复严重周围神经损伤的方案设计提供了实验依据，具有潜在的临床价值。

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