中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (6): 1086-1092.doi: 10.4103/1673-5374.300449

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

改良方法制备异体异种去细胞周围神经移植物修复大鼠坐骨神经缺损

  


  • 出版日期:2021-06-15 发布日期:2020-12-31
  • 基金资助:

    伊朗桂兰医科大学研究与技术研究项目

Decellularized peripheral nerve grafts by a modified protocol for repair of rat sciatic nerve injury

Arash Zaminy1, *, Sara Sayad-Fathi1, Farshad Moharrami Kasmaie2, Zohreh Jahromi2, Adib Zendedel3   

  1. 1 Neuroscience Research Center, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran;  2 Student Research Committee, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran;  3 Institute of Neuroanatomy, Faculty of Medicine, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, Germany 
  • Online:2021-06-15 Published:2020-12-31
  • Contact: Arash Zaminy, PhD, zaminy_a@gums.ac.ir.
  • Supported by:
    This work was fully supported by the Research and Technology Chancellor of Guilan University of Medical Sciences (No. 95110202; to AZa). 

摘要:

研究显示,去细胞神经同种异体移植(ANAs)不易引起免疫排斥反应,用于周围神经缺损的修复较为安全有效;然而,迄今为止,尚无公认的标准化学去细胞方法。研究旨在了解改良的哈德森方法制备的牛源性去细胞神经用于修复大鼠坐骨神经缺损的效果,该方法将Triton X-200替换为Triton X-100,并将DNase和RNase用于去细胞神经移植物的制备中。通过H&E染色,Alcian蓝染色,Masson染色和Luxol固蓝染色,以及免疫组化和生化测定法检测了体外制备的去细胞神经移植物的效率。结果显示,去细胞神经移植物去除了细胞、髓磷脂和轴突,而不影响神经的组织结构。接下来以异体异种去细胞神经移植物桥接大鼠7mm的坐骨神经缺损,以验证修复周围神经缺损的有效性。移植8周后,去细胞神经异种移植修复坐骨神缺损大鼠的坐骨神经功能指数(SFI)与自体神经移植相近;形态学分析显示去细胞神经异种移植物中大量再生的髓鞘化轴突,雪旺细胞数量与自体移植物相当。这些结果说明该改良方法制备的异种异体去细胞移植物可用于修复周围神经缺损。该研究于2017年2月11日获得伊朗桂兰医科大学研究与技术部研究伦理委员会(IR.GUMS.REC.1395.332)的批准。

https://orcid.org/0000-0002-2388-481X (Arash Zaminy)

Abstract: Studies have shown that acellular nerve xenografts do not require immunosuppression and use of acellular nerve xenografts for repair of peripheral nerve injury is safe and effective. However, there is currently no widely accepted standard chemical decellularization method. The purpose of this study is to investigate the efficiency of bovine-derived nerves decellularized by the modified Hudson’s protocol in the repair of rat sciatic nerve injury. In the modified Hudson’s protocol, Triton X-200 was replaced by Triton X-100, and DNase and RNase were used to prepare accelular nerve xenografts. The efficiency of bovine-derived nerves decellularized by the modified Hudson’s protocol was tested in vitro by hematoxylin & eosin, Alcian blue, Masson’s trichrome, and Luxol fast blue staining, immunohistochemistry, and biochemical assays. The decellularization approach excluded cells, myelin, and axons of nerve xenografts, without affecting the organization of nerve xenografts. The decellularized nerve xenograft was used to bridge a 7 mm-long sciatic nerve defect to evaluate its efficiency in the repair of peripheral nerve injury. At 8 weeks after transplantation, sciatic function index in rats subjected to transplantation of acellular nerve xenograft was similar to that in rats undergoing transplantation of nerve allograft. Morphological analysis revealed that there were a large amount of regenerated myelinated axons in acellular nerve xenograft; the number of Schwann cells in the acellular nerve xenograft was similar to that in the nerve allograft. These findings suggest that acellular nerve xenografts prepared by the modified Hudson’s protocol can be used for repair of peripheral nerve injury. This study was approved by the Research Ethics Committee, Research and Technology Chancellor of Guilan  University of Medical Sciences, Iran (approval No. IR.GUMS.REC.1395.332) on February 11, 2017.