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电纺纤维模板光刻法制备的导电和微槽聚合物支架上神经干细胞的培养。

Culture of neural stem cells on conductive and microgrooved polymeric scaffolds fabricated via electrospun fiber-template lithography.

机构信息

Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul, 03722, Republic of Korea.

出版信息

Biomed Mater. 2020 May 19;15(4):045007. doi: 10.1088/1748-605X/ab763b.

DOI:10.1088/1748-605X/ab763b
PMID:32053805
Abstract

We developed polymeric scaffolds that can provide both topographical and electrical stimuli on mouse neural stem cells (mNSCs) for potential use in nerve tissue engineering. In contrast to conventional patterning techniques such as imprinting, soft/photolithography, and three-dimensional printing, microgroove patterns were generated by using aligned electrospun fibers as templates, via a process denoted as electrospun fiber-template lithography. The preparation of polyvinylpyrrolidone fibers, followed by the deposition of poly(lactic-co-glycolic acid) (PLGA) and the removal of the fiber template, produced freestanding PLGA scaffolds with microgrooves having widths of 1.72 ± 0.24 μm. The subsequent deposition of polypyrrole (PPy) via chemical oxidative polymerization added conductivity to the microgrooved PLGA scaffolds. The resultant scaffolds were cytocompatible with mNSCs. The microgroove patterns enhanced the alignment and elongation of mNSCs, and the PPy layer promoted the interaction of cells with the surface by forming more and longer filopodia compared with the nonconductive surface. Finally, the neuron differentiation of mNSCs was evaluated by monitoring the Tuj-1 neuronal gene expression. The presence of both microgrooves and the conductive PPy layer enhanced the neuronal differentiation of mNSCs even without electrical stimulation, and the neuronal differentiation was further enhanced by stimulation with a sufficient electrical pulse (1.0 V).

摘要

我们开发了一种聚合物支架,可对小鼠神经干细胞(mNSC)提供形貌和电刺激,有望用于神经组织工程。与传统的图案化技术(如压印、软光刻和三维打印)不同,微槽图案是通过使用取向的静电纺丝纤维作为模板,通过称为静电纺丝纤维模板光刻的过程产生的。首先制备聚乙烯基吡咯烷酮纤维,然后沉积聚(乳酸-共-乙醇酸)(PLGA)并去除纤维模板,可得到具有 1.72±0.24μm 宽度的独立 PLGA 支架,其上带有微槽。通过化学氧化聚合将聚吡咯(PPy)沉积在微槽 PLGA 支架上,增加了其导电性。所得支架与 mNSC 具有细胞相容性。微槽图案增强了 mNSC 的定向和伸长,与非导电表面相比,PPy 层通过形成更多和更长的丝状伪足促进了细胞与表面的相互作用。最后,通过监测 Tuj-1 神经元基因表达来评估 mNSC 的神经元分化。即使没有电刺激,微槽和导电 PPy 层的存在也增强了 mNSC 的神经元分化,而通过施加足够的电脉冲(1.0 V)刺激则进一步增强了神经元分化。

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