Zaminy Arash, Sayad-Fathi Sara, Kasmaie Farshad Moharrami, Jahromi Zohreh, Zendedel Adib
Neuroscience Research Center, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
Student Research Committee, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
Neural Regen Res. 2021 Jun;16(6):1086-1092. doi: 10.4103/1673-5374.300449.
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.
研究表明,脱细胞神经异种移植物不需要免疫抑制,使用脱细胞神经异种移植物修复周围神经损伤是安全有效的。然而,目前尚无广泛接受的标准化学脱细胞方法。本研究的目的是探讨经改良哈德逊方案脱细胞的牛源神经在大鼠坐骨神经损伤修复中的效率。在改良的哈德逊方案中,用Triton X-100取代了Triton X-200,并使用脱氧核糖核酸酶和核糖核酸酶制备脱细胞神经异种移植物。通过苏木精-伊红染色、阿尔辛蓝染色、马松三色染色、卢克斯尔固蓝染色、免疫组织化学和生化分析,在体外测试经改良哈德逊方案脱细胞的牛源神经的效率。脱细胞方法排除了神经异种移植物中的细胞、髓鞘和轴突,而不影响神经异种移植物的组织结构。将脱细胞神经异种移植物用于桥接7毫米长的坐骨神经缺损,以评估其在周围神经损伤修复中的效率。移植后8周,接受脱细胞神经异种移植物移植的大鼠坐骨神经功能指数与接受神经同种异体移植的大鼠相似。形态学分析显示,脱细胞神经异种移植物中有大量再生的有髓轴突;脱细胞神经异种移植物中雪旺细胞的数量与神经同种异体移植物中的数量相似。这些发现表明,经改良哈德逊方案制备的脱细胞神经异种移植物可用于修复周围神经损伤。本研究于2017年2月11日获得伊朗吉兰医科大学研究与技术副校长研究伦理委员会批准(批准号:IR.GUMS.REC.1395.332)。