中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (3): 573-579.doi: 10.4103/1673-5374.290905

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

硬膜外电刺激可有效恢复完全性脊髓横断损伤大鼠的运动功能

  

  • 出版日期:2021-03-15 发布日期:2020-12-17
  • 基金资助:

    国家自然科学基金（81601052，815201081017）；北京诺华科技计划（2018034）；中国北京市科学技术项（D16100002816005）；中国人民解放军重大项目子公司（AWS17J004）。

Epidural electrical stimulation effectively restores locomotion function in rats with complete spinal cord injury

Song Wang1, 2, Li-Cheng Zhang2, Hai-Tao Fu3, Jun-Hao Deng2, Gao-Xiang Xu2, Tong Li4, Xin-Ran Ji2, *, Pei-Fu Tang2, *   

  1. 1 School of Medicine, Nankai University, Tianjin, China;  2 Department of Orthopedics, Chinese PLA General Hospital, Beijing, China;  3 Department of Orthopedics, The Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, China;  4 Characteristic Medical Center of the Chinese People’s Armed Police Force, Tianjin, China
  • Online:2021-03-15 Published:2020-12-17
  • Contact: Xin-Ran Ji, MD, 18510622211@163.com; Pei-Fu Tang, MD, pftang301@163.com.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, Nos. 81601052 (to XRJ), 81520108017 (to PFT); the Beijing Nova Program of Science and Technology of China, No. 2018034 (to XRJ); the Beijing Municipal Science and Technology Project of China, No. D16100002816005 (to PFT); the Subsidiary of PLA Major Project of China, No. AWS17J004 (to PFT). 

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摘要：

硬膜外电刺激可以通过重新激活残存的神经环路恢复脊髓损伤后的肢体运动功能。既往硬膜外电刺激研究常采用单一电极位点、连续性强直刺激的模式，该刺激条件下机体容易出现耐受性和动作协调性的下降。实验构建完全性脊髓横断伤大鼠模型，在T8水平垂直切断脊髓以排除残存神经纤维对硬膜外电刺激结果的干扰，然后进行硬膜外电刺激，该柔性硬膜外电极具有良好的解剖学拓扑结构能够与植入节段的椎管形态相匹配，同时电极刺激位点可以准确贴服于脊髓L2-L3和S1节段； (1) 为了评估植入硬膜外电刺激电极的生物相容性，在电极植入后第7天对电极下方的脊髓组织取材进行GFAP/Iba-1双标免疫荧光染色显示，电极植入后脊髓局部的小胶质细胞和星形胶质细胞在数量、形态方面无明显增加，未见到活化的Iba-1+细胞聚集，说明植入物不会引起脊髓组织的炎性反应；(2)大鼠步态分析显示，术后第3天，脊髓损伤大鼠在瞬时脉冲刺激下可以在跑步机上恢复协调的步态动作，重建的步态周期可明确区分出支撑相和摆动相并能随刺激发放频率进行动态调整；(3) 为了评估柔性硬膜外电极（内含3个刺激信号发送触点）与椎骨形态的匹配程度及刺激电极的硬膜外位点水平，在电极植入前及植入后均使用Micro-CT对大鼠胸腰椎椎体扫描显示： (4)基于脊髓L2-L3和S1节段的3位点刺激电极联合瞬发性脉冲刺激可以有效恢复SCI大鼠步态动作的实验结果分析，硬膜外电刺激是一种可靠的方法，有待深入探讨其中更多的问题。此项研究于2019年4月19日被中国人民解放军总医院动物伦理委员会批准（批准号：2019-X15-39）。

https://orcid.org/0000-0002-6994-5008 (Xin-Ran Ji); 

https://orcid.org/0000-0003-4279-1704 (Pei-Fu Tang)

关键词: 行为学, 损伤, 模型, 神经功能, 大鼠, 恢复, 再生, 修复, 脊髓

Abstract: Epidural electrical stimulation can restore limb motor function after spinal cord injury by reactivating the surviving neural circuits. In previous epidural electrical stimulation studies, single electrode sites and continuous tetanic stimulation have often been used. With this stimulation, the body is prone to declines in tolerance and locomotion coordination. In the present study, rat models of complete spinal cord injury were established by vertically cutting the spinal cord at the T8 level to eliminate disturbance from residual nerve fibers, and were then subjected to epidural electrical stimulation. The flexible extradural electrode had good anatomical topology and matched the shape of the spinal canal of the implanted segment. Simultaneously, the electrode stimulation site was able to be accurately applied to the L2–3 and S1 segments of the spinal cord. To evaluate the biocompatibility of the implanted epidural electrical stimulation electrodes, GFAP/Iba-1 double-labeled immunofluorescence staining was performed on the spinal cord below the electrodes at 7 days after the electrode implantation. Immunofluorescence results revealed no significant differences in the numbers or morphologies of microglia and astrocytes in the spinal cord after electrode implantation, and there was no activated Iba-1+ cell aggregation, indicating that the implant did not cause an inflammatory response in the spinal cord. Rat gait analysis showed that, at 3 days after surgery, gait became coordinated in rats with spinal cord injury under burst stimulation. The regained locomotion could clearly distinguish the support phase and the swing phase and dynamically adjust with the frequency of stimulus distribution. To evaluate the matching degree between the flexible epidural electrode (including three stimulation contacts), vertebral morphology, and the level of the epidural site of the stimulation electrode, micro-CT was used to scan the thoracolumbar vertebrae of rats before and after electrode implantation. Based on the experimental results of gait recovery using three-site stimulation electrodes at L2–3 and S1 combined with burst stimulation in a rat model of spinal cord injury, epidural electrical stimulation is a promising protocol that needs to be further explored. This study was approved by the Animal Ethics Committee of Chinese PLA General Hospital (approval No. 2019-X15-39) on April 19, 2019. 

Key words: behavior, injury, model, neurological function, rat, recovery, regeneration, repair, spinal cord