Development of three-dimensional DC electric field stimulation bio-reactor for axonal outgrowth enhancement
Development of three-dimensional DC electric field stimulation bio-reactor for axonal outgrowth enhancement
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开发用于增强轴突生长的三维直流电场刺激生物反应器
DOI：
10.1115/imece2018-86637
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Eiji Nakamachi
中科院分区：
文献类型：
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作者：
Shohei Tanaka;Ryota Sakiyama;Koji Yamamoto;Yusuke Morita;Eiji Nakamachi
已经进行了大量关于电刺激对神经再生的影响的研究。然而，采用模拟体内环境的3D培养的研究却很少。在本研究中，我们设计并制造了一种新型 3D 直流电场 (DCEF) 刺激生物反应器，并研究了其增强轴突生长的有效性。我们使用有限元 (FE) 分析寻找最佳结构，以获得培养区域中均匀的 DCEF。 DCEF强度的测量结果与FE结果一致。将大鼠苯色细胞瘤细胞（PC12）散布在胶原凝胶中并进行3D培养。我们使用多光子激发荧光显微镜（MPM）观察细胞体和神经突的形态。在每天6小时、持续4天5.0mV/mm的条件下，平均轴突长度增加了11.3％，轴发生率增加了4.2％。此外，与对照组相比，DCEF组中的细胞体之间的连接距离有更长的趋势。因此，我们使用我们新开发的生物反应器验证了刺激的功效，无论是对于轴突延伸还是神经网络生成。
Numerous studies of electrical stimulation effects on the nerve regeneration have been carried out. However, there were very few investigations which adopt the 3D culture that mimics thein vivoenvironment. In this study, we designed and fabricated a new 3D direct current electric field (DCEF) stimulation bio-reactor and investigated the effectiveness on the axonal outgrowth enhancement. We searched an optimum structure using the finite element (FE) analyses to obtain a uniform DCEF in the culture region. A measurement result of DCEF strength showed an agreement with FE results. The rat phenocromocytoma cells (PC12) were disseminated in the collagen gel and 3D culture was performed. We observed the morphologies of cell bodies and neurites using the multiphoton excitation fluorescence microscope (MPM). Both increases in 11.3% of mean axonal length and in 4.2% of axogenesis rate, under the condition of 5.0 mV/mm on 6 hours a day for 4 days, were obtained. Further, there was a tendency of longer connecting distance between cell bodies in the DCEF group than one in the Control group. As a result, we validated the efficacies of our stimulation, both for the axonal extension and the neural network generation, using our newly developed bio-reactor.