中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (5): 903-912.doi: 10.4103/1673-5374.249239

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

有序纤维神经导管桥接坐骨神经缺损时可提供更好的结构和机械性能

  

  • 出版日期:2019-05-15 发布日期:2019-05-15
  • 基金资助:

    中国国家自然科学基金项目(31771052)；国家重点基础研究发展计划（973计划）(2014CB542201)；中国国家重点研究发展项目(2016YFC1101601, 2017YFA0104702)；解放军总医院转化医学项目(2016TM-030)；北京市自然科学基金项目(7172202)；全军医学青年培养项目(16QNP144)；北京市科技项目(Z161100005016059)

Aligned fibers enhance nerve guide conduits when bridging peripheral nerve defects focused on early repair stage

Qi Quan 1 , Hao-Ye Meng 1, 3 , Biao Chang 2 , Guang-Bo Liu 1 , Xiao-Qing Cheng 1 , He Tang 1 , Yu Wang 1, 4 , Jiang Peng 1, 4 , Qing Zhao 4, 5 , Shi-Bi Lu 1   

  1. 1 Department of Orthopedic Surgery, Key Laboratory of Musculoskeletal Trauma & War Injuries PLA, Beijing Key Lab of Regenerative Medicine in Orthopedics, Chinese PLA General Hospital, Beijing, China
    2 Department of Laser Medicine, Chinese PLA General Hospital, Beijing, China
    3 School of Materials Science and Engineering, University of Science & Technology Beijing, Beijing, China
    4 Neural Regeneration Co-innovation Center of Jiangsu Province, Nantong, Jiangsu Province, China
    5 Department of Orthopedic Surgery, First Affiliated Hospital of PLA General Hospital, Beijing, China
  • Online:2019-05-15 Published:2019-05-15
  • Contact: Shi-Bi Lu, PhD, MD, lusb301@126.com; Qing Zhao, PhD, MD, zqmd301@126.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 31771052 (to YW); the National Basic Re¬search Program of China (973 Program), No. 2014CB542201 (to JP); the National Key Research and Development Program of China, No. 2016YFC1101601 (to QZ), 2017YFA0104702 (to YW); the PLA General Hospital Translational Medicine Project of China, No. 2016TM-030 (to QZ); the Beijing Municipal Natural Science Foundation of China, No. 7172202 (to YW); the PLA Youth Training Project for Medical Sci¬ence, China, No. 16QNP144 (to YW); the Beijing Municipal Science and Technology Project, China, No. Z161100005016059 (to YW).

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摘要：

神经导管在修复长距离周围神经缺损方面有助于神经再生，但构建怎样的神经导管才能更好地发挥这样的功能？实验以多步静电纺丝工艺在神经导管内构建了有序的纳米纤维。(1)以扫描电镜和快速傅里叶变换表征了神经导管内纤维的排列，以机械测试机测试了神经导管在拉伸和抗压性方面的性能；(2)在体外实验中以许旺细胞、PC12细胞和背根神经节与有序纤维神经导管共培养，通过免疫组化染色检测许旺细胞标志物S100的阳性表达以及PC12细胞和背根神经节标志物神经丝蛋白的阳性表达；(3)在体内实验中建立成年雌性SD大鼠右侧坐骨神经1cm缺损模型，以有序纤维神经导管进行桥接，以苏木精-伊红染色和免疫组化观察再生基质，以检测再生基质中ATF3及cleaved caspase-3的阳性表达；(4)在桥接后通过静态坐骨神经指数分析运动功能恢复的情况，以电镜观察远端神经节中有髓纤维数量、轴突直径、纤维直径和髓磷脂厚度，以电子天平检测腓肠肌肌肉质量比；(5)结果发现，有序纤维神经导管具有良好的机械性能，且可诱导许旺细胞、PC12细胞和背根神经节沿纳米纤维方向聚集在纤维周围，并促进后2种细胞长出更长的轴突；在坐骨神经缺损模型大鼠中，有序纤维神经导管能够有效促进ATF3和或cleaved caspase-3阳性表达、坐骨神经功能、腓肠肌的肌肉质量以及远端神经节超微结构的恢复。实验结果显示高度有序排列的纳米纤维在桥接周围神经缺损时可增强神经导管的桥梁性能，增强神经导管在修复周围神经缺损的效果。

orcid: 0000-0003-1493-4042 (Qi Quan)
           0000-0001-9055-6755 (Qing Zhao)

关键词: 神经导管, 多步静电纺丝, 周围神经损伤, 有序纤维, 坐骨神经, 结构, 机械性能, 神经支架, 纳米纤维, 神经再生

Abstract:

Nerve conduits enhance nerve regeneration in the repair of long-distance peripheral nerve defects. To help optimize the effective¬ness of nerve conduits for nerve repair, we developed a multi-step electrospinning process for constructing nerve guide conduits with aligned nanofibers. The alignment of the nerve guide conduits was characterized by scanning electron microscopy and fast Fourier trans¬form. The mechanical performance of the nerve guide conduits was assessed by testing for tensile strength and compression resistance. The biological performance of the aligned fibers was examined using Schwann cells, PC12 cells and dorsal root ganglia in vitro. Immunohisto¬chemistry was performed for the Schwann cell marker S100 and for the neurofilament protein NF200 in PC12 cells and dorsal root ganglia. In the in vivo experiment, a 1.5-cm defect model of the right sciatic nerve in adult female Sprague-Dawley rats was produced and bridged with an aligned nerve guide conduit. Hematoxylin-eosin staining and immunohistochemistry were used to observe the expression of ATF3 and cleaved caspase-3 in the regenerating matrix. The recovery of motor function was evaluated using the static sciatic nerve index. The number of my¬elinated fibers, axon diameter, fiber diameter, and myelin thickness in the distal nerve were observed by electron microscopy. Gastrocnemius muscle mass ratio was also determined. The analyses revealed that aligned nanofiber nerve guide conduits have good mechanical properties and can induce Schwann cells, PC12 cells and dorsal root ganglia to aggregate along the length of the nanofibers, and promote the growth of longer axons in the latter two (neuronal) cell types. The aligned fiber nerve conduits increased the expression of ATF3 and cleaved caspase-3 at the middle of the regenerative matrix and at the distal nerve segment, improved sciatic nerve function, increased muscle mass of the gastroc¬nemius muscle, and enhanced recovery of distal nerve ultrastructure. Collectively, the results show that highly aligned nanofibers improve the performance of the nerve conduit bridge, and enhance its effectiveness in repairing peripheral nerve defects.

Key words: nerve regeneration, nerve guide conduit, electrospinning, peripheral nerve injury, aligned fiber, sciatic nerve, structure, mechanical function, nerve scaffold, nanofiber, neural regeneration