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 (10): 1780-1786.doi: 10.4103/1673-5374.257533

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

骨髓间充质干细胞复合胶原蛋白-壳聚糖支架修复创伤性脑损伤

  

  • 出版日期:2019-10-15 发布日期:2019-10-15
  • 基金资助:

    北京市博士后研究基金（2017-ZZ-120），北京市自然科学基金项目（2164073） ，北京市医院管理局青年计划项目（QML20180804）

Collagen-chitosan scaffold impregnated with bone marrow mesenchymal stem cells for treatment of traumatic brain injury

Feng Yan 1, Ming Li 2, Hong-Qi Zhang 1, Gui-Lin Li 1, Yang Hua 3, Ying Shen 4, Xun-Ming Ji 1, 2, Chuan-Jie Wu 5, Hong An 2, 5, Ming Ren 2   

  1. 1 Department of Neurosurgery, Xuanwu Hospital, Capital Medical University, Beijing, China
    2 China-America Institute of Neuroscience, Xuanwu Hospital, Capital Medical University, Beijing, China
    3 Department of Ultrasound, Xuanwu Hospital, Capital Medical University, Beijing, China
    4 Department of Traditional Chinese Medicine, Xuanwu Hospital, Capital Medical University, Beijing, China
    5 Department of Neurology, Xuanwu Hospital, Capital Medical University, Beijing, China
  • Online:2019-10-15 Published:2019-10-15
  • Contact: Xun-Ming Ji, jixm@ccmu.edu.cn.
  • Supported by:

    This study was financially supported by the Postdoctoral Research Foundation of Beijing of China, No. 2017-ZZ-120 (to FY); the Natural Science Foundation of Beijing of China, No. 2164073 (to ML); the Beijing Municipal Administration of Hospitals’ Youth Plan of China, No. QML20180804 (to ML).

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摘要：

为促进细胞或其他治疗因子向大脑的有效靶向递送并重建受损神经通路，将生物材料与细胞联合治疗脑损伤成为一种有潜力的治疗措施。（1）实验首先基于脑组织工程方法，通过冷冻干燥法制备了胶原蛋白-壳聚糖多孔支架，将其浸润于大鼠骨髓间充质干细胞，制备了骨髓间充质干细胞复合胶原蛋白-壳聚糖支架。然后以300g重物自由落体撞击方法建立创伤性脑损伤大鼠模型，并将骨髓间充质干细胞复合胶原蛋白-壳聚糖支架植入受损脑组织。（2）以改良神经功能严重程度评分评估大鼠的神经功能的恢复情况；以Morris水迷宫检测空间学习和记忆能力；以苏木精-伊红染色检测脑组织病理变化；以免疫组化染色检测血管内皮生长因子的免疫阳性反应以及BrdU/NSE和BrdU/胶质纤维酸性蛋白双阳性反应。（3）结果显示，经骨髓间充质干细胞复合胶原蛋白-壳聚糖支架植入治疗后，创伤性脑损伤大鼠的改良神经功能严重程度评分明显降低，Morris水迷宫平均潜伏期缩短，穿越平台的次数增加，损伤侧脑组织退化细胞减少，缺血移植区及其周围血管内皮生长因子阳性反应增多，移植14d后损伤区骨髓间充质干细胞出现更多的BrdU/胶质纤维酸性蛋白双阳性反应少量BrdU/NSE双阳性反应，且骨髓间充质干细胞复合胶原蛋白-壳聚糖支架植入治疗的效果优于单独骨髓间充质干细胞立体定向注射。（4）为测试骨髓间充质干细胞复合胶原蛋白-壳聚糖支架的生物相容性以及免疫源性，于移植前12h和移植后1-5d静脉注射免疫抑制剂环孢素，可见其与只进行骨髓间充质干细胞复合胶原蛋白-壳聚糖支架修复的大鼠的上述指标接近。（5）表明骨髓间充质干细胞复合胶原蛋白-壳聚糖支架植入治疗可以显著促进创伤性脑损伤大鼠神经功能病理损伤的恢复，可能成为治疗创伤性脑损伤的潜在方法。

orcid:0000-0003-0293-2744 (Xun-Ming Ji)

关键词: 干细胞, 胶原, 壳聚糖, 支架, 创伤性脑损伤, 骨髓间充质干细胞, 脑组织工程, 神经再生

Abstract:

Combinations of biomaterials and cells can effectively target delivery of cells or other therapeutic factors to the brain to rebuild damaged nerve pathways after brain injury. Porous collagen-chitosan scaffolds were prepared by a freeze-drying method based on brain tissue engineering. The scaffolds were impregnated with rat bone marrow mesenchymal stem cells. A traumatic brain injury rat model was estab¬lished using the 300 g weight free fall impact method. Bone marrow mesenchymal stem cells/collagen-chitosan scaffolds were implanted into the injured brain. Modified neurological severity scores were used to assess the recovery of neurological function. The Morris water maze was employed to determine spatial learning and memory abilities. Hematoxylin-eosin staining was performed to measure pathologi¬cal changes in brain tissue. Immunohistochemistry was performed for vascular endothelial growth factor and for 5-bromo-2-deoxyuridine (BrdU)/neuron specific enolase and BrdU/glial fibrillary acidic protein. Our results demonstrated that the transplantation of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds to traumatic brain injury rats remarkably reduced modified neurological severity scores, shortened the average latency of the Morris water maze, increased the number of platform crossings, diminished the degeneration of damaged brain tissue, and increased the positive reaction of vascular endothelial growth factor in the transplantation and surround¬ing areas. At 14 days after transplantation, increased BrdU/glial fibrillary acidic protein expression and decreased BrdU/neuron specific enolase expression were observed in bone marrow mesenchymal stem cells in the injured area. The therapeutic effect of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds was superior to stereotactic injection of bone marrow mesenchymal stem cells alone. To test the biocompatibility and immunogenicity of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds, im¬munosuppressive cyclosporine was intravenously injected 12 hours before transplantation and 1–5 days after transplantation. The above indicators were similar to those of rats treated with bone marrow mesenchymal stem cells and collagen-chitosan scaffolds only. These findings indicate that transplantation of bone marrow mesenchymal stem cells in a collagen-chitosan scaffold can promote the recovery of neuropathological injury in rats with traumatic brain injury. This approach has the potential to be developed as a treatment for traumatic brain injury in humans. All experimental procedures were approved by the Institutional Animal Investigation Committee of Capital Medi¬cal University, China (approval No. AEEI-2015-035) in December, 2015.

Key words: nerve regeneration, stem cells, collagen, chitosan, scaffolds, traumatic brain injury, bone marrow mesenchymal stem cells, brain tissue engineering, neural regeneration