中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (4): 510-515.doi: 10.12307/2022.084

• 药物控释材料 drug delivery materials • 上一篇    下一篇

携带脐带间充质干细胞的RADA16-BDNF水凝胶支架定向移植促进脑出血大鼠神经功能恢复

黄传俊1,2,邹  煜1,周晓婷2,朱扬清1,钱  伟1,张  卫1,刘  星1   

  1. 苏州市第九人民医院(南通大学附属吴江医院),1神经外科,2中心实验室,江苏省苏州市   215000
  • 收稿日期:2020-08-27 修回日期:2020-08-29 接受日期:2020-10-16 出版日期:2022-02-08 发布日期:2021-11-03
  • 通讯作者: 刘星,硕士,副主任医师,苏州市第九人民医院神经外科(南通大学附属吴江医院神经外科),江苏省苏州市 215000
  • 作者简介:黄传俊,男,1989年生,江苏省盐城市人,汉族,2016年南通大学毕业,硕士,主治医师,主要从事脑卒中、颅脑外伤的神经再生基础及临床研究。
  • 基金资助:
    苏州市“科教兴卫”青年科技项目(KJXW2017075),项目负责人:黄传俊;南通大学自然科学类科研基金项目(17ZYZ34),项目负责人:黄传俊;吴江区“科教兴卫”项目(WWWK201913),项目负责人:邹煜;南通大学附属吴江医院科研基金项目(Y201708),项目负责人:黄传俊

Transplantation of umbilical cord mesenchymal stem cells encapsulated in RADA16-BDNF hydrogel promotes neurological recovery in an intracerebral hemorrhage rat model

Huang Chuanjun1, 2, Zou Yu1, Zhou Xiaoting2, Zhu Yangqing1, Qian Wei1, Zhang Wei1, Liu Xing1   

  1. 1Department of Neurosurgery, 2Central Laboratory, Suzhou Ninth People’s Hospital (Wujiang Hospital Affiliated to Nantong University), Suzhou 215000, Jiangsu Province, China
  • Received:2020-08-27 Revised:2020-08-29 Accepted:2020-10-16 Online:2022-02-08 Published:2021-11-03
  • Contact: Liu Xing, Master, Associate chief physician, Department of Neurosurgery, Suzhou Ninth People’s Hospital (Wujiang Hospital Affiliated to Nantong University), Suzhou 215000, Jiangsu Province, China
  • About author:Huang Chuanjun, Master, Attending physician, Department of Neurosurgery, and Central Laboratory, Suzhou Ninth People’s Hospital (Wujiang Hospital Affiliated to Nantong University), Suzhou 215000, Jiangsu Province, China
  • Supported by:
    the Youth Science and Technology Project of “Science, Education, and Health Promotion” of Suzhou, No. KJXW2017075 (to HCJ); Natural Science Research Fund Project of Nantong University, No. 17ZYZ34 (to HCJ); “Science, Education, and Health Promotion” Project of Wujiang, No. WWK201913 (to ZY); Fund Project of Wujiang Hospital Affiliated to Nantong University, No. Y201708 (to HCJ)

摘要:

文题释义:
脑源性神经营养因子:作为在中枢神经系统广泛表达的一种重要神经营养因子,可诱导干细胞神经分化、促进神经细胞存活及生长,对于中枢神经系统损伤及退行性疾病的功能恢复具有重要作用。
水凝胶支架:是一类依靠分子间作用力自发交联成三维孔隙结构的多肽分子,可以模拟细胞外基质给携带的细胞提供支持,促进其增殖、分化,是一种理想的干细胞移植载体,具有广阔的临床应用前景。
背景:脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)在诱导间充质干细胞神经分化及保护内源性神经元存活等方面具有重要作用。课题组在前期研究中构建了RADA16-BDNF多肽水凝胶支架,发现其具有良好的细胞及神经组织相容性,且能促进间充质干细胞的神经分化。
目的:观察携带脐带间充质干细胞的RADA16-BDNF水凝胶支架移植促进脑出血大鼠神经元再生及神经功能恢复的效果及潜在作用机制。
方法:①原子力显微镜、扫描电镜观测RADA16-BDNF水凝胶支架的表面结构及携带脐带间充质干细胞后的表观情况;②构建携带脐带间充质干细胞的RADA16-BDNF水凝胶复合支架,并以脐带间充质干细胞作为对照,体外诱导培养分化,免疫荧光染色观察Ki-67及β微管蛋白Ⅲ的表达及细胞形态;③以自体血立体定向注射法制备大鼠脑出血模型,3 d后定向注射100 µL携带脐带间充质干细胞(PKH26标记)的RADA16-BDNF多肽水凝胶或PBS,移植后不同时间采用水迷宫实验检测大鼠逃逸时间,评估神经功能恢复情况;移植后60 d采用免疫荧光染色观察脑内微管相关蛋白2的表达。
结果与结论:①RADA16-BDNF水凝胶支架具有微孔隙结构,其孔隙可为携带的细胞提供黏附生长的空间;②RADA16-BDNF水凝胶支架能促进脐带间充质干细胞的体外增殖及神经元分化(P < 0.05),其微孔隙结构能促进神经突起的延伸;③RADA16-BDNF水凝胶支架能促进脐带间充质干细胞在大鼠脑内存活并分化为神经元,提高神经功能恢复效果。

https://orcid.org/0000-0002-8950-455X (刘星) 

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: 脐带间充质干细胞, 神经再生, 多肽水凝胶, 脑源性神经营养因子, 脑出血

Abstract: BACKGROUND: Brain-derived neurotrophic factor (BDNF) plays an important role in inducing neural differentiation of mesenchymal stem cells and protecting the survival of endogenous neurons. In our previous study, we constructed the polypeptide hydrogel scaffold, RADA16-BDNF and proved it possess excellent cellular and brain tissue compatibility. The BDNF released from RADA16-BDNF hydrogel may benefit for neural differentiation of mesenchymal stem cells. 
OBJECTIVE: To observe the effect and potential mechanism of transplantation of RADA16-BDNF hydrogel scaffold with mesenchymal stem cells encapsulated for neuronal regeneration and neurological recovery in cerebral hemorrhage rat model. 
METHODS: (1) Atomic force microscope and scanning electron microscope analysis were conducted to observe the surface structure of RADA16-BDNF hydrogel scaffold with or without umbilical cord mesenchymal stem cells. (2) RADA16-BDNF hydrogel with umbilical cord mesenchymal stem cells encapsulated was cultured in vitro, and single umbilical cord mesenchymal stem cell was used as control. Immunofluorescence staining was conducted to detect the expression of Ki-67 and β-Tubulin III and cell morphology. (3) The cerebral hemorrhage rat models were conducted by autologous blood stereotactic injection. RADA16-BDNF polypeptide hydrogel or PBS (100 µL) with umbilical cord mesenchymal stem cells (PKH26 labeled) was injected directionally. The escape time was detected by water maze test at different time after transplantation, and the recovery of neurological function was evaluated. Microtubule-associated protein 2 was detected by immunofluorescence staining 60 days later.
RESULTS AND CONCLUSION: (1) RADA16-BDNF hydrogel scaffold possessed the micro-pore structure and the pores provided a space for adherence and survival for encapsulated cells. (2) RADA16-BDNF hydrogel scaffold promoted the proliferation and neuronal differentiation of umbilical cord mesenchymal stem cells in vitro (P < 0.05), and the micro-pore structure promoted the extension of nerve dendrites. (3) RADA16-BDNF hydrogel scaffolds can promote the umbilical cord mesenchymal stem cells to survive and differentiate into neurons in the rat brain and improve the recovery of neurological function.


Key words: umbilical cord mesenchymal stem cells, neuronal regeneration, polypeptide hydrogel, brain-derived neurotrophic factor, intracerebral hemorrhage

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