摘要
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。
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 a
作者
王梓琪
李晓寅
江继鹏
宋震
李正超
陈淑莲
陈旭义
Wang Ziqi;Li Xiaoyin;Jiang Jipeng;Song Zhen;Li Zhengchao;Chen Shulian;Chen Xuyi(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;General Hospital of Chinese People’s Liberation Army,Beijing 100039,China)
出处
《中国组织工程研究》
CAS
北大核心
2023年第34期5483-5490,共8页
Chinese Journal of Tissue Engineering Research
基金
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。
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
biomedical engineering
collagen/silk fibroin
human umbilical cord-derived mesenchymal stem cell
traumatic brain injury
beagle dog
