中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (11): 1932-1940.doi: 10.4103/1673-5374.259626

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

健侧C7转移联合脱细胞同种异体神经移植修复大鼠臂丛上干神经损伤

  

  • 出版日期:2019-11-15 发布日期:2019-11-15
  • 基金资助:

    国家自然科学基金(81601057)

Contralateral C7 transfer combined with acellular nerve allografts seeded with differentiated adipose stem cells for repairing upper brachial plexus injury in rats

Jian-Tao Yang , Jin-Tao Fang , Liang Li, Gang Chen, Ben-Gang Qin, Li-Qiang Gu   

  1. Department of Microsurgery & Orthopedic Trauma, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
  • Online:2019-11-15 Published:2019-11-15
  • Contact: Li-Qiang Gu, MD, PhD, guliqiang1963@aliyun.com.
  • Supported by:

    This work was supported by the National Natural Science Foundation of China, No. 81601057 (to JTY).

摘要:

健侧C7神经根移位治疗臂丛神经损伤常需要神经移植。去细胞同种异体神经是治疗神经缺损的有效替代材料,复合脂肪干细胞诱导分化的类许旺细胞可进一步提高其修复效果,但其作为桥接神经在臂丛神经损伤中应用的研究尚未见报道。实验以此探讨去细胞同种异体神经复合脂肪干细胞诱导分化的类许旺细胞构建组织工程神经,联合健侧C7神经根移位修复臂丛神经上干损伤的有效性。实验首先提取大鼠脂肪干细胞并采用化学诱导分化为类许旺细胞,取大鼠双侧约15 mm坐骨神经制备同种异体去细胞神经备用。将大鼠随机分为3组,单纯去细胞神经组,复合细胞组(去细胞神经复合类许旺细胞联合移植)及自体神经组,使用开放前入路撕脱臂丛神经根法建立臂丛神经上干损伤大鼠模型,然后行健侧C7神经根经颈部肌肉浅层移位,根据分组选择不同的神经移植体桥接健侧C7神经根及患侧损伤的上干或C5、6神经根。手术后16周采用组织学染色、电生理学检测及神经功能评估神经修复的效果发现:(1)损伤神经功能恢复时间,自体神经组早于复合细胞组,复合细胞组早于单纯去细胞神经组,其中自体神经组和单纯去细胞神经组差异有显著性意义;(2)与单纯去细胞神经组比较,自体神经组和复合细胞组复合肌肉动作电位及运动传导速度、神经移植物内神经元标记物NF及S100阳性率、再生轴突的直径、髓鞘厚度和有髓纤维密度显著增加;(3)上述数据说明,健侧C7转移联合脱细胞同种异体神经移植,可有效促进臂丛上干损伤后的神经修复,效果优于单纯去细胞神经修复。实验经中山大学第一附属医院动物伦理委员会批准(批准号:2016-150)。

orcid: 0000-0003-0532-5093(Li-Qiang Gu)

关键词: 周围神经损伤, 臂丛神经损伤, 健侧颈7神经根, 去细胞神经, 脂肪干细胞, 许旺细胞, 组织工程神经, 神经移植, 神经缺损, 神经再生

Abstract:

Nerve grafting has always been necessary when the contralateral C7 nerve root is transferred to treat brachial plexus injury. Acellular nerve allograft is a promising alternative for the treatment of nerve defects, and results were improved by grafts laden with differentiated adipose stem cells. However, use of these tissue-engineered nerve grafts has not been reported for the treatment of brachial plexus injury. The aim of the present study was to evaluate the outcome of acellular nerve allografts seeded with differentiated adipose stem cells to improve nerve regeneration in a rat model in which the contralateral C7 nerve was transferred to repair an upper brachial plexus injury. Differentiated adipose stem cells were obtained from Sprague-Dawley rats and transdifferentiated into a Schwann cell-like phenotype. Acellular nerve allografts were prepared from 15-mm bilateral sections of rat sciatic nerves. Rats were randomly divided into three groups: acellular nerve allograft, acellular nerve allograft + differentiated adipose stem cells, and autograft. The upper brachial plexus injury model was established by traction applied away from the intervertebral foramen with micro-hemostat forceps. Acellular nerve allografts with or without seeded cells were used to bridge the gap between the contralateral C7 nerve root and C5–6 nerve. Histological staining, electrophysiology, and neurological function tests were used to evaluate the effect of nerve repair 16 weeks after surgery. Results showed that the onset of discernible functional recovery occurred earlier in the autograft group first, followed by the acellular nerve allograft + differentiated adipose stem cells group, and then the acellular nerve allograft group; moreover, there was a significant difference between autograft and acellular nerve allograft groups. Compared with the acellular nerve allograft group, compound muscle action potential, motor conduction velocity, positivity for neurofilament and S100, diameter of regenerating axons, myelin sheath thickness, and density of myelinated fibers were remarkably increased in autograft and acellular nerve allograft + differentiated adipose stem cells groups. These findings confirm that acellular nerve allografts seeded with differentiated adipose stem cells effectively promoted nerve repair after brachial plexus injuries, and the effect was better than that of acellular nerve repair alone. This study was approved by the Animal Ethics Committee of the First Affiliated Hospital of Sun Yat-sen University of China (approval No. 2016-150) in June 2016.

Key words: nerve regeneration, peripheral nerve injury, brachial plexus injury, contralateral C7 nerve root, acellular nerve, adipose stem cells, Schwann cells, tissue engineering nerve, nerve grafting, nerve defect, neural regeneration