中国神经再生研究(英文版) ›› 2016, Vol. 11 ›› Issue (1): 168-173.doi: 10.4103/1673-5374.175065

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

如何能客观全面评估组织工程神经的血管新生?

  

  • 收稿日期:2015-09-23 出版日期:2016-01-15 发布日期:2016-01-15
  • 基金资助:

    中国国家自然科学基金项目(81130080)

Angiogenesis in tissue-engineered nerves evaluated objectively using MICROFIL perfusion and micro-CT scanning

Hong-kui Wang 1, 2, Ya-xian Wang 2, Cheng-bin Xue 2, Zhen-mei-yu Li 2, Jing Huang 2, Ya-hong Zhao 2, Yu-min Yang 2, Xiao-song Gu 1, 2   

  1. 1 School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu Province, China
    2 Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China
  • Received:2015-09-23 Online:2016-01-15 Published:2016-01-15
  • Contact: Xiao-song Gu, M.D.,nervegu@ntu.edu.cn.
  • Supported by:

    study was supported by the National Natural Science Foundation of China, No. 81130080.

摘要:

组织工程神经体内修复过程中良好的血管生成是神经再生的关键因素之一,直接影响着再生的效果。目前为止,还没有一种客观全面且简便可行的组织工程神经血管新生评估方法,直接影响了神经再生研究的进展。实验以体外构建SD大鼠皮肤来源祖细胞分化的许旺细胞为支持细胞、壳聚糖神经导管结合丝素纤维为支架的组织工程神经,桥接修复大鼠10mm坐骨神经缺损。4周后,应用MICROFIL对比剂灌注以及Micro-CT扫描方法,成功地进行了组织工程神经新生血管的三维重建。新生血管由近端、远端和中间3个方向向组织工程神经生长。通过对血管三维图像进行参数分析,获得反映血管的数量、直径、连接和空间分布的参数,且证实新生血管主要是毛细血管和微血管,直径为9-301μm,其中直径为27-155μm占82.84%。说明MICROFIL对比剂灌注以及Micro-CT扫描,是客观全面评估组织工程神经血管新生的简便可行的方法。

关键词: 神经再生, 血管生成, Micro-CT, MICROFIL造影剂, 三维重建, 组织工程神经, 皮肤来源前体细胞, 许旺细胞, 壳聚糖神经导管

Abstract:

Angiogenesis is a key process in regenerative medicine generally, as well as in the specific field of nerve regeneration. However, no convenient
and objective method for evaluating the angiogenesis of tissue-engineered nerves has been reported. In this study, tissue-engineered nerves were constructed in vitro using Schwann cells differentiated from rat skin-derived precursors as supporting cells and chitosan nerve conduits combined with silk fibroin fibers as scaffolds to bridge 10-mm sciatic nerve defects in rats. Four weeks after surgery, three-dimensional blood vessel reconstructions were made through MICROFIL perfusion and micro-CT scanning, and parameter analysis of the tissue-engineered nerves was performed. New blood vessels grew into the tissue-engineered nerves from three main directions: the proximal end, the distal end, and the middle. The parameter analysis of the three-dimensional blood vessel images yielded several parameters, including the number, diameter, connection, and spatial distribution of blood vessels. The new blood vessels were mainly capillaries and microvessels, with diameters ranging from 9 to 301 μm. The blood vessels with diameters from 27 to 155 μm accounted for 82.84% of the new vessels. The microvessels in the tissue-engineered nerves implanted in vivo were relatively well-identified using the MICROFIL perfusion and micro-CT scanning method, which allows the evaluation and comparison of differences and changes of angiogenesis in tissue-engineered nerves implanted in vivo.

Key words: nerve regeneration, angiogenesis, micro-CT, MICROFIL perfusion, three-dimensional reconstruction, tissue-engineered nerve, skin-derived precursor, chitosan nerve conduit, Schwann cell, neural regeneration