中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (34): 5454-5461.doi: 10.3969/j.issn.2095-4344.1445

• 细胞外基质材料 extracellular matrix materials • 上一篇    下一篇

胶原/硫酸肝素支架联合神经干细胞促进脊髓损伤后运动功能的恢复

曹宗锐1，郑  博1，钟  琳2，胡良聪3，张秀莉1，屈  波1，蒋  涛1
  

  1. 1成都医学院第一附属医院骨科，四川省成都市  610500；2昆明医科大学第二附属医院，云南省昆明市  650101；3华中科技大学同济医学院附属协和医院骨科，湖北省武汉市  430022
  • 收稿日期:2019-05-22 出版日期:2019-12-08 发布日期:2019-12-08
  • 通讯作者: 蒋涛，主任医师，副教授，成都医学院第一附属医院骨科，四川省成都市 610500
  • 作者简介:曹宗锐，男，1982年生，四川省达州市人，主治医师，讲师，主要从事关节、骨质疏松和脊柱疾病研究。
  • 基金资助:

    四川省教育厅支撑项目(15AZ0265)，项目名称：老年男性髋部骨折患者血清睾酮水平与骨质疏松、骨折风险的相关性，项目负责人：蒋涛

Collagen/heparin sulfate scaffold combined with neural stem cells promote motor function recovery after spinal cord injury

Cao Zongrui1, Zheng Bo1, Zhong Lin2, Hu Liangcong3, Zhang Xiuli1, Qu bo1, Jiang Tao1
  

  1. 1Department of Orthopedics, First Affiliated Hospital of Chengdu Medical College, Chengdu 610500, Sichuan Province, China; 2Second Affiliated Hospital of Kunming Medical University, Kunming 650101, Yunan Province, China; 3Department of Orthopedics, Affiliated Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
  • Received:2019-05-22 Online:2019-12-08 Published:2019-12-08
  • Contact: Jiang Tao, Chief physician, Associate professor, Department of Orthopedics, First Affiliated Hospital of Chengdu Medical College, Chengdu 610500, Sichuan Province, China
  • About author:Cao Zongrui, Attending physician, Lecturer, Department of Orthopedics, First Affiliated Hospital of Chengdu Medical College, Chengdu 610500, Sichuan Province, China
  • Supported by:

    Sichuan Provincial Department of Education-Supported Project, No. 15AZ0265 (to JT)

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摘要：

文章快速阅读：

 

文题释义：
硫酸肝素：是在所有动物组织中发现的线性多糖，为一种蛋白多糖，其中2个或3个硫酸肝素链紧密附着在细胞表面或细胞外基质蛋白附近，硫酸肝素结合大量细胞外蛋白质并调节多种生物活性。
弥散张量成像：是一种描述脊髓结构的新方法，是核磁共振成像(MRI)的特殊形式。举例来说，如果说核磁共振成像是追踪水分子中的氢原子，那么弥散张量成像便是依据水分子移动方向制图。弥散张量成像图(呈现方式与以前的图像不同)可以揭示脑瘤如何影响神经细胞连接，引导医疗人员进行脊髓手术，其还可以揭示同脑卒中、多发性硬化症、精神分裂症、阅读障碍有关的细微反常变化。
 
 
背景：近些年来，胶原/硫酸肝素支架作为神经组织工程支架已被用于周围神经、脊髓和脑损伤的修复中。
目的：观察胶原/硫酸肝素支架联合神经干细胞移植对脊髓损伤后运动功能恢复的影响。
方法：将SD乳鼠神经干细胞接种于胶原/硫酸肝素支架上培养7 d，构建细胞-支架复合物。将60只成年雌性SD大鼠(购自成都达硕生物科技有限公司)随机分为4组，每组15只：假手术组仅切除T10椎板，模型组切除T10处1.5 mm脊髓建立脊髓损伤模型，支架组建立脊髓损伤模型后植入胶原/硫酸肝素支架，细胞-支架组建立脊髓损伤模型后植入细胞-支架复合物。术后1，2，3，4，6，8周进行BBB评分与斜板实验，观察大鼠肢体功能恢复；术后8周检测大鼠双后肢运动诱发电位，观察运动功能恢复情况；术后8周对脊髓损伤处进行核磁扫描，获得弥散张量成像图像，观察神经纤维再生情况。实验获得成都医学院第一附属医院伦理委员会批准。
结果与结论：①术后2，3，4，6，8周，细胞-支架组的BBB评分、斜板实验角度均高于模型组、支架组  (P < 0.05，P < 0.01)；②细胞-支架组大鼠双侧后肢的运动诱发电位振幅高于模型组、支架组(P < 0.05，P < 0.01)，潜伏期短于模型组、支架组(P < 0.05，P < 0.01)；③弥散张量成像显示，假手术组神经纤维完整，模型组病变部位周围神经纤维缺乏不同方向的连续性，支架组、细胞-支架组可见较多数量的神经纤维穿过损伤区域，并且细胞-支架组穿过损伤区域的神经纤维数量更多；④结果表明，胶原/硫酸肝素支架联合神经干细胞可促进大鼠脊髓损伤处神经纤维的再生，改善大鼠双侧后肢运动功能。

关键词: 脊髓损伤, 胶原, 硫酸肝素, 胶原/硫酸肝素支架, 支架, 神经干细胞, 神经再生, 功能恢复, 弥散张量纤维束成像

Abstract:

Abstract
BACKGROUND: In recent years, collagen/heparin sulfate scaffolds have been used as nerve tissue engineering scaffolds in the repair of peripheral nerves, spinal cords, and brain injury.
OBJECTIVE: To investigate the effects of collagen/heparin sulfate scaffold combined with neural stem cells on motor function recovery after spinal cord injury.
METHODS: Sprague-Dawley rat neural stem cells were inoculated on a collagen/heparin sulfate scaffold for 7 days to construct a cell-scaffold complex. Sixty adult female Sprague-Dawley rats (purchased from Chengdu Dashuo Biotechnology Co., Ltd., China) were randomly divided into four groups with 15 rats in each group. In the sham operation group, only T10 lamina was removed. In the model group, rat models of spinal cord injury were stablished by removing 1.5 mm long spinal cord at the level of T10 lamina. In the scaffold group, collagen/heparin sulfate scaffolds were implanted in the spinal cord injury sites. In the cell-scaffold complex group, collagen/heparin sulfate scaffold-neural stem cells complex was implanted into the injury site. At postoperative 1, 2, 3, 4, 6 and 8 weeks, BBB score and oblique plate test were performed to evaluate the recovery of limb function in rats. At postoperative 8 weeks, the motor evoked potentials of rat hind limbs were detected and the recovery of motor function was evaluated. At postoperative 8 weeks, a nuclear magnetic scan was performed on the spinal cord injury site, and a diffusion tensor imaging image was obtained to observe nerve fiber regeneration. This study was approved by the Ethics Committee of the First Affiliated Hospital of Chengdu Medical College, China.
RESULTS AND CONCLUSION: At postoperative 2, 3, 4, 6 and 8 weeks, the BBB score and oblique plate test angle in the cell-scaffold complex group were significantly greater than those in the model and scaffold groups (P < 0.05, P < 0.01). The amplitude of motor evoked potential in the rat hind limbs in the cell-scaffold group was significantly higher than that in the model and scaffold groups (P < 0.05, P < 0.01). The latency in the cell-scaffold group was significantly shorter than that in the model and scaffold groups (P < 0.05, P < 0.01). Diffusion tensor imaging revealed that nerve fibers were intact in the sham operation group and the nerve fibers around the lesions in the model group lacked continuity in different directions. In the scaffold and cell-scaffold groups, a large number of nerve fibers passed through the lesion area. The number of nerve fibers passing through the lesion area in the cell-scaffold group was higher than that in the scaffold group. These results suggest that collagen/heparin sulfate scaffold combined with neural stem cells can promote the regeneration of nerve fibers in the spinal cord injury area and improve the motor function of rat hind limbs.

Key words: spinal cord injury, collagen, heparin sulfate, collagen/heparin sulfate scaffold, neural stem cells, nerve regeneration, functional recovery, diffusion tensor

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