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层粘连蛋白包被的静电纺丝再生丝素蛋白基质促进神经祖细胞增殖、分化和存活

Laminin-Coated Electrospun Regenerated Silk Fibroin Mats Promote Neural Progenitor Cell Proliferation, Differentiation, and Survival .

作者信息

Li Guangfei, Chen Kai, You Dan, Xia Mingyu, Li Wen, Fan Suna, Chai Renjie, Zhang Yaopeng, Li Huawei, Sun Shan

机构信息

NHC Key Laboratory of Hearing Medicine, State Key Laboratory of Medical Neurobiology, Shanghai Engineering Research Centre of Cochlear Implant, Otorhinolaryngology Department of Affiliated Eye and ENT Hospital, Ear, Nose & Throat Institute, Fudan University, Shanghai, China.

State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, International Joint Laboratory for Advanced Fiber and Low-Dimension Materials, College of Materials Science and Engineering, Donghua University, Shanghai, China.

出版信息

Front Bioeng Biotechnol. 2019 Aug 6;7:190. doi: 10.3389/fbioe.2019.00190. eCollection 2019.

Abstract

Neural progenitor cell (NPC) transplantation is a promising technique for central nervous system (CNS) reconstruction and regeneration. Biomaterial scaffolds, frameworks, and platforms can support NPC proliferation and differentiation as well as serve as a temporary extracellular matrix after transplantation. However, further applications of biomaterials require improved biological attributes. Silk fibroin (SF), which is produced by , is a widely used and studied protein polymer for biomaterial application. Here, we prepared aligned and random eletrospun regenerated SF (RSF) scaffolds, and evaluated their impact on the growth of NPCs. First, we isolated NPCs and then cultured them on either laminin-coated RSF mats or conventional laminin-coated coverslips for cell assays. We found that aligned and random RSF led to increases in NPC proliferation of 143.8 ± 13.3% and 156.3 ± 14.7%, respectively, compared to controls. Next, we investigated neuron differentiation and found that the aligned and the random RSF led to increases in increase in neuron differentiation of about 93.2 ± 6.4%, and 3167.1 ± 4.8%, respectively, compared to controls. Furthermore, we measured the survival of NPCs and found that RSF promoted NPC survival, and found there was no difference among those three groups. Finally, signaling pathways in cells cultured on RSF mats were studied for their contributions in neural cell differentiation. Our results indicate that RSF mats provide a functional microenvironment and represent a useful scaffold for the development of new strategies in neural engineering research.

摘要

神经祖细胞(NPC)移植是一种用于中枢神经系统(CNS)重建和再生的很有前景的技术。生物材料支架、框架和平台可以支持NPC的增殖和分化,并且在移植后可作为临时的细胞外基质。然而,生物材料的进一步应用需要改善其生物学特性。由蚕产生的丝素蛋白(SF)是一种广泛用于生物材料应用且经过研究的蛋白质聚合物。在此,我们制备了排列型和随机型的静电纺丝再生丝素蛋白(RSF)支架,并评估了它们对NPC生长的影响。首先,我们分离出NPC,然后将它们培养在层粘连蛋白包被的RSF垫或传统的层粘连蛋白包被的盖玻片上进行细胞检测。我们发现,与对照组相比,排列型和随机型RSF分别使NPC增殖增加了143.8±13.3%和156.3±14.7%。接下来,我们研究了神经元分化,发现与对照组相比,排列型和随机型RSF分别使神经元分化增加了约93.2±6.4%和3167.1±4.8%。此外,我们测量了NPC的存活率,发现RSF促进了NPC的存活,并且发现这三组之间没有差异。最后,研究了在RSF垫上培养的细胞中的信号通路对神经细胞分化的作用。我们的结果表明,RSF垫提供了一个功能性微环境,是神经工程研究中开发新策略的一种有用支架。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f7/6691020/8bbb736cd6ed/fbioe-07-00190-g0001.jpg

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