丝素蛋白/病毒各向异性复合支架的构筑及其在神经修复中的应用

doi:10.19894/j.issn.1000-0518.210290

摘要/Abstract

摘要： 神经修复支架是解决神经损伤的有效途径之一,三维各向异性的支架可以更好地模仿天然细胞外基质从而顺利桥接受损神经。本研究首先利用单向冷冻技术构筑了具有多通道结构的三维各向异性丝素蛋白(SF)支架,并且通过改变丝素蛋白自组装时间和冷冻剂温度调节了孔道的形貌结构,制备出孔径大小为(140±79) μm的适宜支架基底。然后利用“灌注-冻干”法,在SF支架孔道壁表面引入含有促细胞黏附精氨酸-甘氨酸-天冬氨酸(RGD)肽的烟草花叶病毒突变体(TMV-RGD1)。体外实验表明,神经细胞能够良好地粘附在SF/TMV-RGD1复合支架上,并且其轴突可以沿孔道纵向生长。研究证明,通过在SF支架孔道壁表面引入TMV-RGD1,一方面在化学层面引入了RGD活性序列,另一方面在物理层面丰富了基底的各向异性纳米拓扑结构,改善了SF基底的细胞亲和性并促进了细胞轴突的定向生长。

关键词: 丝素蛋白, 烟草花叶病毒, 各向异性, 神经修复

Abstract: Nerve repair scaffolds are one of the effective ways to solve nerve damage. Three-dimensional anisotropic scaffolds can better mimic the natural extracellular matrix to smoothly bridge the damaged nerves. In this study, a three-dimensional anisotropic silk fibroin (SF) scaffold with a multi-channel structure was constructed using unidirectional freezing technology, and the pore morphology and structure of the channels were adjusted by changing the self-assembly time of silk fibroin and the temperature of the refrigerant to prepare a suitable scaffold substrate with a pore size of (140±79) μm. Then, using the “perfusion-freeze-drying” method, the tobacco mosaic virus mutant (TMV-RGD1, RGD: Arg-Gly-Asp) was introduced on the surface of the SF scaffold hole. In vitro experiments show that nerve cells can adhere well to the SF/TMV-RGD1 composite scaffold, and their axons can grow longitudinally along the pores. It has been proved that by introducing TMV-RGD on the surface of the pores of the SF scaffold, on the one hand, the active sequence of RGD is introduced at the chemical level, on the other hand, the anisotropic nanotopological structure of the substrate is enriched at the physical level. The cell affinity of the SF substrate is improved and the directional growth of cell axons is promoted.

Key words: Silk fibroin, Tobacco mosaic virus, Anisotropy, Neural repair

中图分类号:

O636

孔倩, 林园, 苏朝晖. 丝素蛋白/病毒各向异性复合支架的构筑及其在神经修复中的应用[J]. 应用化学, 2021, 38(11): 1454-1461.

KONG Qian, LIN Yuan, SU Zhao-Hui. Construction of Silk Fibroin/Virus Anisotropic Scaffold and Its Application in Nerve Repair[J]. Chinese Journal of Applied Chemistry, 2021, 38(11): 1454-1461.

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