中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (5): 959-968.doi: 10.4103/1673-5374.268974

• 原著:脊髓损伤修复保护与再生 • 上一篇    

3-D生物打印胶原/丝素蛋白支架联合神经干细胞促进脊髓损伤后神经再生

  

  • 出版日期:2020-05-15 发布日期:2020-06-01

Three-dimensional bioprinting collagen/silk fibroin scaffold combined with neural stem cells promotes nerve regeneration after spinal cord injury

Ji-Peng Jiang1, 2, Xiao-Yin Liu2, 3, Fei Zhao2, Xiang Zhu4, Xiao-Yin Li2, Xue-Gang Niu5, Zi-Tong Yao2, Chen Dai2, Hui-You Xu2, Ke Ma2, Xu-Yi Chen2, Sai Zhang2#br#   

  1. 1 Department of Thoracic Surgery, General Hospital of People’s Liberation Army (PLA), Beijing, China
    2 Tianjin Key Laboratory of Neurotrauma Repair, Institute of Traumatic Brain Injury and Neuroscience, Center for Neurology and Neurosurgery of Chinese People’s Armed Police Force (PAP) Medical Center, Tianjin, China
    3 Tianjin Medical University, Tianjin, China
    4 Department of Neurology, Luoyang First Hospital of Traditional Chinese Medicine, Luoyang, Henan Province, China
    5 Department of Neurosurgery, Fourth Central Hospital of Tianjin, Tianjin, China
  • Online:2020-05-15 Published:2020-06-01
  • Contact: Xu-Yi Chen,chenxuyi1979@126.com; Sai Zhang,zhangsai718@vip.126.com.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. 11672332 (to XYC); the National Key Research and Development Plan of China, No. 2016YFC1101500 (to SZ).

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摘要：

已有大量研究表明,生物材料支架在促进损伤脊髓轴突再生等方面具you1重要价值。细胞移植和生物材料支架植入被认为是用于神经组织修复的有效方法。为此实验作了如下设计：(1)制造备了一种三维胶原/丝素蛋白支架(3D-CF)，其腔体模拟正常脊髓的解剖结构，允许细胞在体外和体内生长，观察其联合神经干细胞(NSCs)移植对脊髓损伤修复的作用；(2)将40只SD大鼠随机分成4组，假手术组仅进行椎板切除术，脊髓损伤组以T10脊髓全横断建立脊髓损伤模型，3D-CF组脊髓损伤后损伤脊髓组织植入3D-CF支架，3D-CF+NSCs组脊髓损伤后损伤脊髓组织采用3D-CF支架和神经干细胞移植联合干预；(3)以神经电生理检测、影像学检测、苏木精伊红染色、嗜银染色、免疫荧光染色和蛋白质印迹分析进行效果评估，与假手术组之外的其他各组相比，3D-CF+NSCs组的神经行为学评分明显升高，电生理检测中运动诱发电位潜伏期明显缩短，振幅明显增大，MRI和DTI显示的脊髓连续性最好，脊髓损伤空腔填充最好，可见大量再生的神经纤维和轴突，几乎无胶质瘢痕的增生；(4)上述数据证实，实验所制备的3-D生物打印胶原/丝素蛋白支架联合神经干细胞移植可促进大鼠脊髓损伤的修复。实验经中国人民武装警察部队特色医学中心动物伦理委员会批准(批准号：2017-0007.2)。

orcid: 0000-0002-0420-8349 (Xu-Yi Chen) 

         0000-0002-8028-4183 (Sai Zhang)

关键词: 脊髓损伤, 胶原, 丝素蛋白, 支架, 3D生物打印, 神经干细胞, 神经再生, 功能恢复, 磁共振成像, 扩散张量成像

Abstract: Many studies have shown that bio-scaffolds have important value for promoting axonal regeneration of injured spinal cord. Indeed, cell transplantation and bio-scaffold implantation are considered to be effective methods for neural regeneration. This study was designed to fabricate a type of three-dimensional collagen/silk fibroin scaffold (3D-CF) with cavities that simulate the anatomy of normal spinal cord. This scaffold allows cell growth in vitro and in vivo. To observe the effects of combined transplantation of neural stem cells (NSCs) and 3D-CF on the repair of spinal cord injury. Forty Sprague-Dawley rats were divided into four groups: sham (only laminectomy was performed), spinal cord injury (transection injury of T10 spinal cord without any transplantation), 3D-CF (3D scaffold was transplanted into the local injured cavity), and 3D-CF + NSCs (3D scaffold co-cultured with NSCs was transplanted into the local injured cavity. Neuroelectrophysiology, imaging, hematoxylin-eosin staining, argentaffin staining, immunofluorescence staining, and western blot assay were performed. Apart from the sham group, neurological scores were significantly higher in the 3D-CF + NSCs group compared with other groups. Moreover, latency of the 3D-CF + NSCs group was significantly reduced, while the amplitude was significantly increased in motor evoked potential tests. The results of magnetic resonance imaging and diffusion tensor imaging showed that both spinal cord continuity and the filling of injury cavity were the best in the 3D-CF + NSCs group. Moreover, regenerative axons were abundant and glial scarring was reduced in the 3D-CF + NSCs group compared with other groups. These results confirm that implantation of 3D-CF combined with NSCs can promote the repair of injured spinal cord. This study was approved by the Institutional Animal Care and Use Committee of People’s Armed Police Force Medical Center in 2017 (approval No. 2017-0007.2).

Key words: 3D bioprinting, collagen, diffusion tensor imaging, functional recovery, magnetic resonance imaging, nerve regeneration, neural regeneration, neural stem cell, scaffold, silk fibroin, spinal cord injury