中国组织工程研究

中国组织工程研究 ›› 2021, Vol. 25 ›› Issue (16): 2514-2520.doi: 10.3969/j.issn.2095-4344.3116

• 复合支架材料 composite scaffold materials • 上一篇    下一篇

康复训练结合神经营养因子3-壳聚糖支架改善脊髓损伤大鼠骨骼肌形态和运动功能

陈星颖1，郝  飞2，3，高钰丹1，赵  文1，段红梅1，杨朝阳1，李晓光1，2，3   

  1. 1首都医科大学神经生物学系，北京市  100069；北京航空航天大学，2生物与医学工程学院，生物材料和神经再生北京市重点实验室，3医工交叉创新研究院，生物医学工程高精尖创新中心，北京市   100083
  • 收稿日期:2020-03-17 修回日期:2020-03-28 接受日期:2020-06-03 出版日期:2021-06-08 发布日期:2021-01-07
  • 通讯作者: 李晓光，博士，教授，首都医科大学神经生物学系，北京市 100069；北京航空航天大学生物与医学工程学院，生物材料和神经再生北京市重点实验室，北京市 100083；北京航空航天大学医工交叉创新研究院，生物医学工程高精尖创新中心，北京市 100083
  • 作者简介:陈星颖，女，1994年生，福建省福州市人，首都医科大学在读硕士，主要从事应用活性生物材料支架体结合康复训练对于功能恢复影响的相关研究。
  • 基金资助:
    国家自然科学基金(81941011，31730030，31650001，31320103903)，项目负责人：李晓光；国家重点研发计划(2017YFC1104002，2017YFC1104001)，项目负责人：李晓光；国家自然科学基金(31670988，31971279)，项目负责人：杨朝阳；北京市科技计划(Z181100001818007)，项目负责人：杨朝阳；北京市教育委员会2018年度科技计划重点项目(KZ201810025030)，项目负责人：杨朝阳；国家自然科学基金(31771053)，项目负责人：段红梅；中国博士后科学基金面上项目(230210465)，项目负责人：郝飞

Rehabilitation training combined with neurotrophin 3-chitosan scaffolds enhanced skeletal muscle morphology and functional recovery in rats with spinal cord injury

Chen Xingying1, Hao Fei2, 3, Gao Yudan1, Zhao Wen1, Duan Hongmei1, Yang Chaoyang1, Li Xiaoguang1, 2, 3#br#

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  1. 1Department of Neurobiology, Capital Medical University, Beijing 100069, China; 2Beijing Key Laboratory of Biomaterials and Nerve Regeneration, School of Biological Science and Medical Engineering, Beihang University, Beijing 100083, China; 3Biomedical Engineering Innovation Center, Medical Industry Cross Innovation Research Institute, Beihang University, Beijing 100083, China 
  • Received:2020-03-17 Revised:2020-03-28 Accepted:2020-06-03 Online:2021-06-08 Published:2021-01-07
  • Contact: Li Xiaoguang, MD, Professor, Department of Neurobiology, Capital Medical University, Beijing 100069, China; Beijing Key Laboratory of Biomaterials and Nerve Regeneration, School of Biological Science and Medical Engineering, Beihang University, Beijing 100083, China; Biomedical Engineering Innovation Center, Medical Industry Cross Innovation Research Institute, Beihang University, Beijing 100083, China
  • About author:Chen Xingying, Master candidate, Department of Neurobiology, Capital Medical University, Beijing 100069, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81941011, 31730030, 31650001, 31320103903 (to LXG); the National Key Research and Development Program, No. 2017YFC1104002, 2017YFC1104001 (to LXG); the National Natural Science Foundation of China, No. 31670988, 31971279 (to YCY); the Beijing Science and Technology Plan, No. Z181100001818007 (to YCY); the Key Project of Pcience and Technology Plan of Beijing Municipal Education Commission in 2018, No. KZ201810025030 (to YCY); the National Natural Science Foundation of China, No. 31771053 (to DHM); the General Program of China Postdoctoral Science Foundation, No. 230210465 (to HF)

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摘要：

文题释义：
活性生物材料支架：神经营养因子3-壳聚糖活性生物材料支架，可以在体外缓慢释放神经营养因子3达14周，具有良好的生物活性和组织相容性，能为体外细胞培养及体内细胞生存提供良好的细胞外微环境。

背景：前期研究显示，神经营养因子3(neurotrophin 3，NT3)-壳聚糖可诱发脊髓损伤大鼠内源性神经发生和轴突再生，促进大鼠运动和感觉功能恢复。
目的：观察康复训练结合NT3-壳聚糖活性生物材料支架对完全性脊髓损伤大鼠骨骼肌形态变化和功能恢复的影响。
方法：将50只成年雌性Wistar大鼠随机分为5组，每组10只：假手术组不造模，其余4组制备T7-T8全切5 mm脊髓损伤模型，单损组造模后不进行任何干预，另3组分别给予康复训练、NT3-壳聚糖活性生物材料支架、NT3-壳聚糖活性生物材料支架结合康复训练干预，康复训练于造模后2周开始。造模前及造模后2，4，6，8，10，12周对各组大鼠进行开放场地的BBB评分；造模后12周，取后肢骨骼肌(胫骨前肌、腓肠肌、比目鱼肌)进行苏木精-伊红和乙酰胆碱酯酶染色，评定各组大鼠肌肉萎缩和运动终板的变化情况。实验方案经首都医科大学动物实验委员会批准(批准号为AEEI-2018-105)。
结果与结论：①假手术组术后各时间点的BBB评分高于其他4组(P < 0.05)，NT3-壳聚糖结合康复训练组造模后8，10，12周的评分高于单损组、单损结合康复训练组、NT3-壳聚糖组(P < 0.05)；②造模后12周苏木精-伊红染色显示，造模4组的各骨骼肌肌纤维横截面积和直径小于假手术组(P < 0.05)，其中NT3-壳聚糖结合康复训练组各骨骼肌肌纤维横截面积和直径大于单损组、单损结合康复训练组、NT3-壳聚糖组(P < 0.05)；③造模后12周乙酰胆碱酯酶染色显示，造模4组各骨骼肌的运动终板乙酰胆碱酯酶平均吸光度值均低于假手术组(P < 0.05)，其中NT3-壳聚糖结合康复训练组高于单损组、单损结合康复训练组、NT3-壳聚糖组(P < 0.05)；④结果表明，康复训练结合NT3-壳聚糖活性生物材料支架植入能有效防止完全性脊髓损伤大鼠后肢骨骼肌肌肉萎缩，提高运动终板乙酰胆碱酯酶活性，减轻神经肌肉接头退变，改善大鼠后肢运动功能。

https://orcid.org/0000-0002-6928-2718(陈星颖) 

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

关键词: 材料, 神经营养因子3, 脊髓损伤, 生物活性支架, 康复训练, 运动终板, 骨骼肌

Abstract: BACKGROUND: Previous studies have shown that neurotrophin 3 (NT3)-chitosan can induce endogenous neurogenesis and axon regeneration in rats with spinal cord injury, and promote recovery of motor and sensory functions in rats. 
OBJECTIVE: To observe the effect of rehabilitation training combined with NT3-chitosan biomaterial scaffold on skeletal muscle morphological changes and functional recovery in rats with complete spinal cord injury.
METHODS: Fifty adult female Wistar rats were randomly divided into five groups, 10 in each group. The sham group was not modeled; the remaining four groups were prepared with T7-T8 complete 5-mm spinal cord injury model, and the lesion control was not performed any intervention after modeling. The other three groups were given rehabilitation training, NT3-chitosan active biomaterial scaffold, NT3- chitosan active biomaterial scaffold combined with rehabilitation training intervention. Rehabilitation training started 2 weeks after modeling. Before operation, 2, 4, 6, 8, 10, and 12 weeks after operation, all of rats were subjected to double-blind open-field BBB scores. After 12 weeks, the skeletal muscles of the hind limbs (tibialis anterior muscle, gastrocnemius muscle, and soleus muscle) were taken for hematoxylin-eosin staining and acetylcholinesterase staining. The changes in muscle atrophy and motor endplates were assessed in each group. The experimental plan was approved by the Animal Experiment Committee of Capital Medical University (approval No. AEEI-2018-105).
RESULTS AND CONCLUSION: (1) The BBB score at each time point in the sham group was higher than that in the other four groups (P < 0.05); and the scores at 8, 10, and 12 weeks after the NT3-chitosan combined rehabilitation training group were higher than the lesion control group, the lesion control combined rehabilitation training group, and NT3-chitosan group (P < 0.05). (2) At 12 weeks after operation, hematoxylin-eosin staining showed that the cross-sectional area and diameter of muscle fibers of each skeletal muscle were smaller in the other four groups than that in the sham group (P < 0.05). The cross-sectional area and diameter of muscle fibers of each skeletal muscle in the NT3-chitosan combined rehabilitation training group were higher than the lesion control group, the lesion control combined rehabilitation training group, and NT3-chitosan group (P < 0.05). (3) At 12 weeks after operation, the acetylcholinesterase staining showed that the average optical density of the acetylcholinesterase on motor endplate of the muscle was lower in the other four groups than that in the sham group (P < 0.05); the average optical density of the acetylcholinesterase of the motor endplate in the NT3-chitosan combined rehabilitation training was significantly higher than that in the lesion control, lesion control combined rehabilitation training, and NT3-chitosan groups (P < 0.05). (4) The results show that NT3-chitosan combined with rehabilitation training can effectively prevent muscular atrophy of hind limb skeletal muscles in rats with complete spinal cord injury, improve the average optical density of the acetylcholinesterase of the motor endplate, reduce neuromuscular joint degeneration, and improve rat hindlimb motor function. 

Key words: material, neurotrophic factor 3, spinal cord injury, bioactive scaffold, rehabilitation training, motor endplate, skeletal muscle

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