中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (8): 686-692.doi: 10.3969/j.issn.1673-5374.2013.08.002

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

石英晶体微天平快速准确鉴别脊神经前后根

  

  • 收稿日期:2012-12-13 修回日期:2013-02-01 出版日期:2013-03-15 发布日期:2013-03-15
  • 基金资助:

    国家自然科学基金面上项目（No. 30973058，81171694）；省科技厅社会发展计划（BE2010743）；南京医科大学第一附属医院创新团队（No. IRT-015），江苏省优势学科项目。

Rapid identification of spinal ventral and dorsal roots using a quartz crystal microbalance

Tao Sui1, Jun Que2, Dechao Kong1, Hao Xie1, Daode Wang1, Kun Shi1, Xiaojian Cao1, Xiang Li1   

  1. 1 Department of Orthopedics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China
    2 Department of Intensive Care Unit, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China
  • Received:2012-12-13 Revised:2013-02-01 Online:2013-03-15 Published:2013-03-15
  • Contact: Xiaojian Cao, M.D., Chief physician, Department of Orthopedics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China, xiaojiancao@ gmail.com. Xiang Li, M.D., Chief physician, Department of Orthopedics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China, lixiangnjgk@163.com.
  • About author:Tao Sui☆, M.D. Tao Sui and Jun Que contributed equally to this work.
  • Supported by:

    This work was supported by the National Natural Science Foundation of China, No. 30973058, 81171694; Jiangsu Province Natural Science Foundation, No. BE2010743; Jiangsu Graduate Student Innovation Project, No.CXZZ11_0721; the Program for Development of Innovative Research Team in the First Affiliated Hospital of Nanjing Medical University, No. IRT-015; and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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摘要：

神经束的快速、准确鉴别是实现正确吻合的关键。依据脊神经前后根内乙酰胆碱酯酶含量不同的事实，课题组提出了一种基于乙酰胆碱酯酶抗体的石英晶体微天平法。此方法是将乙酰胆碱酯酶抗体包被于石英晶体微天平电极表面，并与样品溶液中的乙酰胆碱酯酶发生抗原抗体反应，在电极表面形成抗原抗体复合物，增加电极质量，导致其频率发生改变，从而实现脊神经前后根的快速、准确鉴别。为验证其效果，实验将犬的脊神经前后根取出，切成1-2mm的片段，浸泡于250 μL PBS中，同时将乙酰胆碱酯酶单克隆抗体固定于石英晶体微天平电极表面。结果显示，在10min内，脊神经前后根均能引起电极频率的改变，且前根明显高于后根。同时对牛血清白蛋白和单纯的PBS溶液的检测均未见电极频率变化。说明石英晶体微天平是一种快速、高灵敏度、高特异性、操作方便的鉴别断裂脊神经前后根的方法。

关键词: 神经再生, 周围神经损伤, 石英晶体微天平, 生物传感器, 脊神经, 神经束, 乙酰胆碱酯酶, 抗原抗体反应, 鉴别, 吻合, 神经修复, 基金资助文章, 图片文章

Abstract:

The fast and accurate identification of nerve tracts is critical for successful nerve anastomosis. Taking advantage of differences in acetylcholinesterase content between the spinal ventral and dorsal roots, we developed a novel quartz crystal microbalance method to distinguish between these nerves based on acetylcholinesterase antibody reactivity. The acetylcholinesterase antibody was immobilized on the electrode surface of a quartz crystal microbalance and reacted with the acetylcholinesterase in sample solution. The formed antigen and antibody complexes added to the mass of the electrode inducing a change in frequency of the electrode. The spinal ventral and dorsal roots were distinguished by the change in frequency. The ventral and dorsal roots were cut into 1 to 2-mm long segments and then soaked in 250 μL PBS. Acetylcholinesterase antibody was immobilized on the quartz crystal microbalance gold electrode surface. The results revealed that in 10 minutes, both spinal ventral and dorsal roots induced a frequency change; however, the frequency change induced by the ventral roots was notably higher than that induced by the dorsal roots. No change was induced by bovine serum albumin or PBS. These results clearly demonstrate that a quartz crystal microbalance sensor can be used as a rapid, highly sensitive and accurate detection tool for the quick identification of spinal nerve roots intraoperatively.

Key words: neural regeneration, peripheral nerve injury, basic research, quartz crystal microbalance, biosensor, spinal nerve, nerve tract, acetylcholinesterase, antigen-antibody reaction, identification, anastomosis, nerve repair, grants-supported paper, neuroregeneration