中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (6): 1093-1098.doi: 10.4103/1673-5374.300463

• 原著:退行性病与再生 • 上一篇    下一篇

冻干或静电纺丝制备壳聚糖支架谁更适合周围神经再生?

  

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

    基金资助:国家自然科学基金(30973060)

Production of chitosan scaffolds by lyophilization or electrospinning: which is better for peripheral nerve regeneration?

Yu-Xuan Wu1, #, Hao Ma2, #, Jian-Lan Wang3, Wei Qu1, *   

  1. 1 Department of Hand Microsurgery, First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning Province, China;  2 Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China;  3 Department of Traditional Chinese Medicine, First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning Province, China
  • Online:2021-06-15 Published:2020-12-31
  • Contact: Wei Qu, PhD, dlquweidoc@hotmail.com.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. 30973060 (to WQ).

摘要:

冷冻干燥和静电纺丝技术是制备壳聚糖支架的常用方法,但是哪种方法制备的支架更适合细胞生长,目前少有研究。实验以冷冻干燥法制备壳聚糖支架与出生3-5 d的SD乳鼠坐骨神经许旺细胞共培养设为冷冻干燥组,以静电纺丝方法制备壳聚糖支架与许旺细胞共培养设为静电纺丝组,以在培养皿上正常培养的许旺细胞设为对照组。通过免疫荧光和扫描电子显微镜分别观察各组许旺细胞的生长状态变化,采用蛋白质印迹实验以探索许旺细胞生长的机制发现:(1)两种材料均无毒且适宜许旺细胞生长;(2)静电纺丝的纳米网状纤维结构比冻干支架的孔隙更小;(3)静电纺丝组中许旺细胞的增殖率和黏附率均优于冻干组;(4)在静电纺丝支架上培养的许旺细胞呈线性结构生长,类似于Burger band,并且分泌了更多的脑源性神经营养因子,从而促进神经元的生长;(5)上述数据揭示,静电纺丝方法制备的壳聚糖支架具有纳米纤维结构,为细胞间的相互作用提供了优良的细胞外基质;相比于冷冻干燥法,静电纺丝方法制备的壳聚糖支架更加适合周围神经的再生。

https://orcid.org/0000-0002-0069-7472 (Wei Qu) 

关键词: 细胞, 因子, 生长, 体外, 模型, 周围神经, 可塑性, 蛋白质, 大鼠, 再生

Abstract: Both lyophilization and electrospinning are commonly used to make chitosan scaffolds. However, it remains unknown which method is better for cell growth. In this study, we established the following groups: (1) lyophilization group—chitosan scaffolds were prepared by lyophilization method and seeded with Schwann cells from Sprague-Dawley rats aged 3–5 days; (2) electrospinning group—chitosan scaffolds were prepared by electrospinning method and seeded with Schwann cells; (3) control group—Schwann cells were cultured on culture dishes. The growth of Schwann cells was evaluated by immunofluorescence and scanning electron microscopy. Western blot assay was performed to explore the mechanism of Schwann cell growth. Both materials were non-toxic and suitable for the growth of Schwann cells. The pores produced by electrospinning were much smaller than those produced by lyophilization. The proliferation rate and adhesion rate of Schwann cells in the electrospinning group were higher than those in the lyophilization group. Schwann cells cultured on electrospinning scaffolds formed a Bungner band-like structure, and a much greater amount of brain-derived neurotrophic factor was secreted, which can promote the growth of neurons. Our findings show that the chitosan scaffold prepared by the electrospinning method has a nanofiber structure that provides an extracellular matrix that is more favorable for cell-cell interactions. The electrospinning method is more suitable for nerve regeneration than the lyophilization method. This research was approved by the Medical Ethical Committee of Dalian Medical University (approval No. AEE1-2016-045) on March 3, 2016.

Key words: cells, factor, growth, in vitro, model, peripheral nerve, plasticity, protein, rat, regeneration