中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (34): 5483-5490.doi: 10.12307/2023.725

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

胶原/丝素蛋白支架联合人脐带源间充质干细胞干预犬创伤性脑损伤

王梓琪1，2，李晓寅2，江继鹏3，宋  震1，2，李正超2，陈淑莲2，陈旭义2   

  1. 1天津中医药大学研究生院，天津市   301617；2中国人民武装警察部队特色医学中心海上维权医学保障研究所，天津市神经创伤修复重点实验室，天津市   300162；3中国人民解放军总医院，北京市   100039
  • 收稿日期:2022-10-13 接受日期:2022-11-18 出版日期:2023-12-08 发布日期:2023-04-22
  • 通讯作者: 陈旭义，医学博士，研究员，博士生导师，中国人民武装警察部队特色医学中心海上维权医学保障研究所，天津市神经创伤修复重点实验室，天津市 300162
  • 作者简介:王梓琪，女，1993年生，湖北省武汉市人，汉族，天津中医药大学在读硕士，主要从事中医药、中西医结合防治脑病与生物组织工程等方面的研究。 李晓寅，男，1986 年生，天津市人，汉族，2014 年天津中医药大学毕业，硕士，研究实习员，主要从事神经病学、生物组织工程、人工智能等方面的研究。
  • 基金资助:
    国家科技重点研发计划(2016YFC1101500)，项目参与者：陈旭义；国防科技卓越青年科学基金(2021-JCJQ-ZQ-035)，项目负责人：陈旭义；武警特色医学中心科研创新团队(KYCXTD0104)，课题负责人：李正超

Collagen/silk fibroin scaffold combined with human umbilical cord-derived mesenchymal stem cells in the treatment of traumatic brain injury in dogs

Wang Ziqi1, 2, Li Xiaoyin2, Jiang Jipeng3, Song Zhen1, 2, Li Zhengchao2, Chen Shulian2, Chen Xuyi2   

  1. 1Graduate School of Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China; 2Research Institute of Maritime Rights Defense Medical Support, Chinese People’s Armed Police Force Characteristic Medical Center, Tianjin Key Laboratory of Neurotrauma Repair, Tianjin 300162, China; 3General Hospital of Chinese People’s Liberation Army, Beijing 100039, China
  • Received:2022-10-13 Accepted:2022-11-18 Online:2023-12-08 Published:2023-04-22
  • Contact: Chen Xuyi, MD, Researcher, Doctoral supervisor, Research Institute of Maritime Rights Defense Medical Support, Chinese People’s Armed Police Force Characteristic Medical Center, Tianjin Key Laboratory of Neurotrauma Repair, Tianjin 300162, China
  • About author:Wang Ziqi, Master candidate, Graduate School of Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China; Research Institute of Maritime Rights Defense Medical Support, Chinese People’s Armed Police Force Characteristic Medical Center, Tianjin Key Laboratory of Neurotrauma Repair, Tianjin 300162, China Li Xiaoyin, Master, Research intern, Research Institute of Maritime Rights Defense Medical Support, Chinese People’s Armed Police Force Characteristic Medical Center, Tianjin Key Laboratory of Neurotrauma Repair, Tianjin 300162, China
  • Supported by:
    National Science and Technology Key Research & Development Plan, No. 2016YFC1101500 (to CXY); National Defense Science and Technology Excellence Youth Science Fund, No. 2021-JCJQ-ZQ-035 (to CXY); Scientific Research and Innovation Team of Armed Police Characteristic Medical Center, No. KYCXTD0104 (to LZC)

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摘要：


文题释义：

扩散张量纤维束成像：是利用弥散张量成像中得出的数据直观地描述白质纤维束分布的技术，其前提是纤维素走行方向与扩散本征矢量供线方向上的最大本征系数方向一致。脑白质纤维素示踪图是目前唯一能在活体显示白质纤维素走行方向的成像技术。
RNA原位杂交：杂交组织化学或细胞的杂交，是一种能够从形态学上证明特异性RNA序列存在于制备的个别细胞、组织部分、单细胞或染色体中的技术，可提供RNA在组织细胞中的空间表达信息。

背景：迄今为止，创伤性脑损伤颠覆性的治疗手段非常有限，医学与工程的结合为中枢神经系统神经修复与再生带来了新的前景。
目的：探讨可携带种子细胞、兼具安全性与多孔隙的胶原/丝素蛋白支架联合人脐带间充质干细胞对犬创伤性脑损伤的治疗作用。
方法：①将第3代人脐带间充质干细胞接种至胶原/丝素蛋白支架上，倒置相差显微镜、扫描电镜、免疫荧光染色、苏木精-伊红染色观察人脐带间充质干细胞生长情况。②将24只比格犬利用随机数字法分为4组：创伤组只建立创伤性脑损伤模型不做其他处理，干细胞组造模后移植人脐带间充质干细胞，支架组造模后移植胶原/丝素蛋白支架，联合组造模后移植人脐带间充质干细胞与胶原/丝素蛋白支架，移植物均在损伤灶局部移植。术后1 d以及1，4，8，12，16，20，24周分别进行改良格拉斯哥昏迷评分评估，术后1，3，6个月进行运动诱发电位检测，术后6个月行核磁共振成像弥散张量成像和脑组织修复RNA原位杂交，评估创伤性脑损伤恢复情况。

结果与结论：①人脐带间充质干细胞贴附胶原/丝素蛋白支架表面生长良好并伸出许多伪足；②术后1，4，8，12，16，20，24周，联合组改良格拉斯哥评分显著高于创伤组、干细胞组、支架组(P < 0.05)；③不同恒压刺激下，1，3，6个月时联合组运动诱发电位潜伏期、振幅均极其显著优于创伤组(P < 0.01)；④核磁共振成像弥散张量成像显示：联合组皮质脊髓束完整性好于创伤组、干细胞组和支架组，损伤侧可见皮质新生；⑤脑组织修复RNA原位杂交结果：联合组Syn、MAP-2、vWF、NEFM、MBP表达量多于创伤组、干细胞组和支架组；⑥结果表明：人脐带间充质干细胞联合胶原/丝素蛋白支架对犬创伤性脑损伤后皮质脊髓束新生、运动功能恢复、血管新生与突起生长发挥了积极作用。

https://orcid.org/0000-0002-0420-8349 (陈旭义) 

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: 生物医学工程, 胶原/丝素蛋白, 人脐带源间充质干细胞, 创伤性脑损伤, 比格犬

Abstract: BACKGROUND: So far, the subversive treatment of traumatic brain injury is very limited. The combination of medicine and engineering has brought new prospects for the repair and regeneration of central nervous system nerves.  
OBJECTIVE: To investigate the therapeutic effect of collagen/silk fibroin scaffold that can carry seed cells and is safe and porous, combined with human umbilical cord-derived mesenchymal stem cells on traumatic brain injury in beagle dogs.
METHODS: (1) Passage 3 human umbilical cord-derived mesenchymal stem cells were inoculated onto collagen/silk fibroin scaffold. The growth of human umbilical cord-derived mesenchymal stem cells was observed by inverted phase contrast microscope, scanning electron microscope, immunofluorescence staining and hematoxylin-eosin staining. (2) Twenty-four beagle dogs were randomly divided into four groups. In the trauma group, only the traumatic brain injury model was established without other treatment. In the stem cell group, human umbilical cord-derived mesenchymal stem cells were transplanted after model establishment. In the scaffold group, the collagen/silk fibroin scaffold was transplanted after model establishment. In the combination group, human umbilical cord-derived mesenchymal stem cells and collagen/silk fibroin scaffold were transplanted after model establishment. The grafts were all transplanted locally in the injured area. At 1 day, 1, 4, 8, 12, 16, 20 and 24 weeks after operation, a modified Glasgow Coma Scale was performed respectively. Motor-evoked potentials were detected at 1, 3 and 6 months after operation. At 6 months after operation, magnetic resonance imaging and in situ hybridization of brain tissue repair RNA were performed to evaluate the recovery of traumatic brain injury.  
RESULTS AND CONCLUSION: (1) Human umbilical cord-derived mesenchymal stem cells attached to collagen/silk fibroin scaffold surface grew well and extended many pseudopods. (2) Modified Glasgow Coma Scale score of the combination group was significantly higher than that of the trauma group, stem cell group and scaffold group at 1, 4, 8, 12, 16, 20 and 24 weeks after operation (P < 0.05). (3) The latency and amplitude of motor-evoked potential of the combination group at 1, 3 and 6 months were significantly better than those of the trauma group under different constant pressure stimuli (P < 0.01). (4) Diffusion tensor imaging of magnetic resonance scanning showed that the integrity of the corticospinal tract in the combination group was better than that in the trauma, stem cell and scaffold groups, and a new corticospinal tract could be seen on the injured side. (5) The results of in situ hybridization of brain tissue repair RNA demonstrated that the expression levels of synaptophysin, microtubule-associated protein 2, von Willebrand factor, neurofilament protein and MBP in the combination group were more than those in the trauma, stem cell and scaffold groups. (6) It is indicated that human umbilical cord-derived mesenchymal stem cells combined with collagen/silk fibroin scaffold play an active role in corticospinal tract regeneration, motor function recovery, angiogenesis and neurite growth in dogs with traumatic brain injury.

Key words: biomedical engineering, collagen/silk fibroin, human umbilical cord-derived mesenchymal stem cell, traumatic brain injury, beagle dog

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