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利用导电和不导电的微纤维支架进行细胞生长和神经发生的电刺激。

Electrical stimulation of cell growth and neurogenesis using conductive and nonconductive microfibrous scaffolds.

机构信息

Faculty of Science, Engineering and Technology, Swinburne University of Technology, PO Box 218, Hawthorn, Melbourne, Australia.

出版信息

Integr Biol (Camb). 2019 Jun 1;11(6):264-279. doi: 10.1093/intbio/zyz022.

Abstract

The effect of exogenous electrical stimulation on cell viability, attachment, growth, and neurogenesis was examined using PC12 cells in microfibrous viscose-rayon scaffolds immersed in culture medium. The scaffolds were applied either in their nonconductive state or after coating the fibres with 200 nm of gold to give a scaffold sheet resistivity of (13 ± 1.3) Ω square-1. The cells were treated for 12 days using direct current electrical stimulation of 2 h per day. No cytotoxic effects were observed when up to 500 mV (8.3 mV mm-1) was applied to the scaffolds without gold, or when up to 100 mV (1.7 mV mm-1) was applied to the scaffolds with gold. Compared with unstimulated cells, whereas electrical stimulation significantly enhanced cell growth and attachment in the nonconductive scaffolds without gold, similar effects were not found for the conductive scaffolds with gold. Neural differentiation in the presence of nerve growth factor was improved by electrical stimulation in both scaffolds; however, neurite development and the expression of key differentiation markers were greater in the nonconductive scaffolds without gold than in the scaffolds with gold. Application of the same current to scaffolds with and without gold led to much higher levels of neurogenesis in the scaffolds without gold. This work demonstrates that substantial benefits in terms of cell growth and neural differentiation can be obtained using electric fields exerted across nonconductive microfibrous scaffolds, and that this approach to electrical stimulation can be more effective than when the stimulus is applied to cells on conductive scaffolds.

摘要

采用浸于培养基中的 PC12 细胞,研究了外加电刺激对细胞活力、黏附、生长和神经发生的影响。这些支架应用于非导电状态或纤维涂覆 200nm 金后,使支架的片电阻(13±1.3)Ω/square-1。采用每天 2 小时的直流电刺激对细胞进行 12 天处理。当未涂金的支架施加高达 500mV(8.3mV·mm-1)或涂金的支架施加高达 100mV(1.7mV·mm-1)时,观察到无细胞毒性作用。与未刺激的细胞相比,尽管在没有金的非导电支架中,电刺激显著增强了细胞的生长和黏附,但在具有金的导电支架中未发现类似的效果。在神经生长因子存在下的神经分化通过两种支架中的电刺激得到改善;然而,在没有金的非导电支架中,神经突发育和关键分化标志物的表达比具有金的支架中更大。相同电流施加到有金和无金的支架上,导致无金支架中的神经发生水平更高。这项工作表明,在非导电微纤维支架上施加电场可以获得细胞生长和神经分化方面的显著益处,并且这种电刺激方法比将刺激施加到导电支架上的细胞时更有效。

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