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 (1): 152-161.doi: 10.4103/1673-5374.264462

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

骨骼肌衍生细胞修复小鼠周围神经缺损

  

  • 出版日期:2020-01-15 发布日期:2020-05-20
  • 基金资助:
    国家自然科学基金(81671908，81571921)；中央大学基础研究基金(2016ZX310197)；中国青年科学与研究基金会(3332015155); 中国医学科学院整形外科医院科学基金(Q2015013)

Skeletal muscle-derived cells repair peripheral nerve defects in mice

Zi-Xiang Chen1, Hai-Bin Lu1, Xiao-Lei Jin1, Wei-Feng Feng2, Xiao-Nan Yang1, Zuo-Liang Qi1   

  1. 1 The 16th Department, Plastic Surgery Hospital, Chinese Academy of Medical Sciences (CAMS) and Peking Union Medical College (PUMC), Beijing, China
    2 Yu Tian Cheng Plastic Surgery Clinic, Shanghai, China
  • Online:2020-01-15 Published:2020-05-20
  • Contact: Xiao-Nan Yang, MD, PhD,yxnan@aliyun.com; Zuo-Liang Qi, MD, PhD,public_qi@163.com.
  • Supported by:
    This study was financially supported by the National Natural Science Foundation of China, No. 81671908 (to ZLQ) and No. 81571921 (to XNY); the Fundamental Research Fund for the Central Universities of China, No. 2016ZX310197 (to ZLQ); the Union Youth Science & Research Foundation of China, No. 3332015155 (to XNY); the Science Fund of Plastic Surgery Hospital, Chinese Academy of Medical Sciences, and Peking Union Medical College of China, No. Q2015013 (to XNY).

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摘要：

肌肉中的骨骼肌源性细胞具有强大的分泌功能，同时其所包含的骨骼肌来源干细胞具有向许旺细胞样细胞等细胞分化的功能，但目前仍未见采用骨骼肌源性细胞治疗周围神经缺损的报道。为此，实验将小鼠右侧坐骨神经被切除5 mm以构建周围神经缺损模型，实验分4组，骨骼肌源性细胞组用聚氨酯导管桥接缺损神经断端，在导管内注入骨骼肌源性细胞进行移植修复；骨骼肌神经导管组和聚氨酯组分别用骨骼肌神经导管和聚氨酯导管桥接缺损神经断端修复；假手术组只分离神经周围组织但不切除神经。术后8，12周，通过免疫荧光染色观察神经内轴突和髓鞘的分布情况以及计算神经的直径；通过甲苯胺蓝染色和透射电镜计算有髓神经纤维的数目、直径和髓鞘厚度；通过腓肠肌切片的Masson染色来计算肌纤维面积占比。术后4，8，12周通过步态足迹分析计算坐骨功能指数。结果显示，(1)术后8，12周，与聚氨酯组比较，骨骼肌来源细胞组的髓鞘和轴突均匀分布，有髓神经纤维数目、直径及髓鞘厚度、腓肠肌湿质量及肌纤维面积比显著增高；(2)术后4，8，12周，骨骼肌源性细胞组坐骨功能指数均显著高于聚氨酯组；(3)在上述全部检测指标中，骨骼肌源性细胞组与骨骼肌神经导管组的检测结果无显著差异；(4)以上结果表明，骨骼肌源性细胞可有效促进小鼠周围神经损伤后的再生。

orcid: 0000-0002-3335-7669 (Xiao-Nan Yang)
         0000-0002-8730-065X (Zuo-Liang Qi)

关键词: 骨骼肌来源细胞, 周围神经再生, 周围神经修复, 骨骼肌, 骨骼肌神经导管, 神经缺损, 种子细胞, 聚氨酯导管, 肌肉因子, 组织工程神经

Abstract: Skeletal muscle-derived cells have strong secretory function, while skeletal muscle-derived stem cells, which are included in muscle-derived cells, can differentiate into Schwann cell-like cells and other cell types. However, the effect of muscle-derived cells on peripheral nerve defects has not been reported. In this study, 5-mm-long nerve defects were created in the right sciatic nerves of mice to construct a peripheral nerve defect model. Adult female C57BL/6 mice were randomly divided into four groups. For the muscle-derived cell group, muscle-derived cells were injected into the catheter after the cut nerve ends were bridged with a polyurethane catheter. For external oblique muscle-fabricated nerve conduit and polyurethane groups, an external oblique muscle-fabricated nerve conduit or polyurethane catheter was used to bridge the cut nerve ends, respectively. For the sham group, the sciatic nerves on the right side were separated but not excised. At 8 and 12 weeks post-surgery, distributions of axons and myelin sheaths were observed, and the nerve diameter was calculated using immunofluorescence staining. The number, diameter, and thickness of myelinated nerve fibers were detected by toluidine blue staining and transmission electron microscopy. Muscle fiber area ratios were calculated by Masson’s trichrome staining of gastrocnemius muscle sections. Sciatic functional index was recorded using walking footprint analysis at 4, 8, and 12 weeks after operation. The results showed that, at 8 and 12 weeks after surgery, myelin sheaths and axons of regenerating nerves were evenly distributed in the muscle-derived cell group. The number, diameter, and myelin sheath thickness of myelinated nerve fibers, as well as gastrocnemius muscle wet weight and muscle area ratio, were significantly higher in the muscle-derived cell group compared with the polyurethane group. At 4, 8, and 12 weeks post-surgery, sciatic functional index was notably increased in the muscle-derived cell group compared with the polyurethane group. These criteria of the muscle-derived cell group were not significantly different from the external oblique muscle-fabricated nerve conduit group. Collectively, these data suggest that muscle-derived cells effectively accelerated peripheral nerve regeneration. This study was approved by the Animal Ethics Committee of Plastic Surgery Hospital, Chinese Academy of Medical Sciences (approval No. 040) on September 28, 2016.

Key words: muscle nerve conduit, myokine, nerve regeneration, nerve repair, peripheral nerve regeneration, polyurethane catheter, seed cells,
skeletal muscle, skeletal muscle-derived cells, tissue-engineered nerve