An in Vitro Stretch-Injury Model for Elongation-Controlled Neuronal Cells: Effect of Strains Along Neurite
An in Vitro Stretch-Injury Model for Elongation-Controlled Neuronal Cells: Effect of Strains Along Neurite
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伸长控制神经元细胞的体外拉伸损伤模型：沿神经突的应变效应
DOI：
10.1007/978-3-319-02913-9_194
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
A.
中科院分区：
文献类型：
--
作者：
Nakadate;H.;Aomura;S. and Kakuta;A.
弥漫性轴索损伤(DAI)是创伤性脑损伤的主要组成部分，与脑组织的快速变形导致神经轴突的伸展有关。局灶性轴索串珠是DAI病理的形态特征，导致神经元与组织的脱节，导致细胞死亡。我们的目标是更好地了解神经元耐受，并有助于预测DAI从机械负荷到头部的发病机制。在本研究中，我们建立了一种牵张损伤模型，利用微流控培养技术控制轴突的方向对培养的神经细胞进行单轴拉伸，并检测沿轴突的应变对细胞损伤的影响。使用具有微槽的聚二甲基硅氧烷(PDMS)片与聚二甲基硅氧烷(PDMS)基板相结合，将PC 12细胞的轴突从0、45和90度延伸到拉伸方向。在应变为0.22、应变率为27 S− 1的条件下，在5min~24 h内观察到相同轴突的形态，结果显示，在拉伸后即刻，沿0度方向的轴突珠数增加，并持续到24 h，而沿45°和90度方向的轴突珠数在拉伸后1 h内有一过性增加。0度和90度方向的轴突较45度方向的轴突破裂更多，但轴突的回缩和消失在任何方位条件下均无差异。这些结果表明，轴突上应变的不同会导致不同类型和程度的轴突损伤，而不是细胞培养底物的应变，而是轴突上加载的应变对神经元损伤的评估是重要的。
Diffuse axonal injury (DAI), a major component of traumatic brain injury, is associated with rapid deformation of brain tissue resulting in the stretching of neural axons. Focal axonal beading, which is the morphological hallmarks of DAI pathology, leads to the disconnection of neurons from tissues, resulting in cell death. Our goal is better understanding of neuronal tolerance and help to predicting the pathogenesis of DAI from mechanical loading to the head. In present study, we developed a stretch-injury model that subjected cultured neuronal cells in which the directions of neurite elongation were controlled with microfluidic culture technique to uniaxial stretch and examined the effect of strains along neurite on the cell damage. Neurites from PC 12 cells were extended at 0, 45 and 90 degrees to stretch direction using a fabricated poly(dimethylsiloxane) (PDMS) piece having microgrooves in combination with PDMS substrate. Following stretch with a strain of 0.22 and a strain rate of 27 s− 1, the morphology of same neurites were observed at 5 min-24 h. As a result, the beading along neurites oriented at 0 degree increased immediately following stretch and the increase was sustained until 24 h, although the beading along neurites oriented at 45 and 90 degrees transiently increased within 1 h following stretch. Many rupture of neurites was observed more in neurites oriented at 0 and 90 degrees than in neurites oriented at 45 degrees, however, the retraction and disappearance of neurites did not differ in any orientation conditions. These results suggest that the difference in strain along neurite induces different types and degrees of neurite damage and not strain of cellcultured substrate but strain loaded to neurite is important to evaluation of neuronal injury.