Electrochemically induced in vitro focal hypoxia in human neurons.
Electrochemically induced in vitro focal hypoxia in human neurons.
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DOI：
10.3389/fcell.2022.968341
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发表时间：
2022
影响因子：
5.5
通讯作者：
Hall, Elizabeth A. H.
中科院分区：
文献类型：
--
作者：
Wong, Joseph J. Y.;Varga, Balazs V.;Karadottir, Ragnhildur Thora;Hall, Elizabeth A. H.
关键词：
局灶性缺氧在中风、心脏骤停和痴呆等疾病中广泛存在。虽然在某些情况下缺氧可以改善细胞功能，但它主要会诱发或加剧病理变化。缺乏可以在动物或细胞培养物中模拟局灶性急性缺氧的方法，阻碍了我们对缺氧细胞后果的理解。为了解决这一差距，报道了一种电化学局部除氧系统（eLOS），为体外时空缺氧调节提供了一个创新平台。电化学系统的建模显示了 O2 通量模式以及局部 O2 清除和缺氧区域，作为距电极和周围通量屏障的距离的函数，从而可以为体外细胞培养研究设计有效的局部缺氧工具。根据 O2 通量边界，电化学定义的目标区域中的 O2 浓度在约 6 分钟内从常氧降至缺氧。因此，设计了一个细胞培养孔，可以在其中诱导局部氧气清除。局部缺氧对人类神经祖细胞 (hNPC) 的影响得到了证实，并且可以诱导微小的局灶性缺氧损伤，从而引起时间依赖性 HIF-1α 转录因子的积累。这种转录根据电化学诱导的时空缺氧梯度在整个培养物中“模式化”。通过在专门设计的微流体装置中应用 eLOS，还开发了基本的腔隙性梗塞模型。在完全氧合的细胞体（例如人类皮质神经元的轴突）的细胞过程中诱导了微型局灶性缺氧损伤。结果通过实验证明，尽管神经元仍处于常氧状态，但局部轴突缺氧应激可导致神经元死亡显着增加。这表明，仅对轴突的局灶性缺氧损伤就足以影响周围的神经元，并可能提供体外模型来研究大脑深部白质中发生的微梗死的影响，并为更广泛地了解急性缺氧损伤提供有前途的工具，并有可能揭示其在多种疾病中的病理生理学。
Focalised hypoxia is widely prevalent in diseases such as stroke, cardiac arrest, and dementia. While in some cases hypoxia improves cellular functions, it mostly induces or exacerbates pathological changes. The lack of methodologies that can simulate focal acute hypoxia, in either animal or cell culture, impedes our understanding of the cellular consequences of hypoxia. To address this gap, an electrochemical localised oxygen scavenging system (eLOS), is reported, providing an innovative platform for spatiotemporal in vitro hypoxia modulation. The electrochemical system is modelled showing O2 flux patterns and localised O2 scavenging and hypoxia regions, as a function of distance from the electrode and surrounding flux barriers, allowing an effective focal hypoxia tool to be designed for in vitro cell culture study. O2 concentration is reduced in an electrochemically defined targeted area from normoxia to hypoxia in about 6 min depending on the O2-flux boundaries. As a result, a cell culture-well was designed, where localised O2 scavenging could be induced. The impact of localised hypoxia was demonstrated on human neural progenitor cells (hNPCs) and it was shown that miniature focal hypoxic insults can be induced, that evoke time-dependent HIF-1α transcription factor accumulation. This transcription is “patterned” across the culture according to the electrochemically induced spatiotemporal hypoxia gradient. A basic lacunar infarct model was also developed through the application of eLOS in a purpose designed microfluidic device. Miniature focal hypoxic insults were induced in cellular processes of fully oxygenated cell bodies, such as the axons of human cortical neurons. The results demonstrate experimentally that localised axonal hypoxic stress can lead to significant increase of neuronal death, despite the neurons remaining at normoxia. This suggests that focal hypoxic insult to axons alone is sufficient to impact surrounding neurons and may provide an in vitro model to study the impact of microinfarcts occurring in the deep cerebral white matter, as well as providing a promising tool for wider understanding of acute hypoxic insults with potential to uncover its pathophysiology in multiple diseases.
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影响因子：
29.4
作者：
Cummins, Eoin P.;Seeballuck, Fergal;Taylor, Cormac T.
通讯作者：
Taylor, Cormac T.
影响因子：
6.6
作者：
Islam, Md. Merajul;Kant, Rama
通讯作者：
Kant, Rama
影响因子：
8
作者：
HUANG, LK;WANG, MJJ
通讯作者：
WANG, MJJ
影响因子：
15.9
作者：
Imtiyaz, Hongxia Z.;Williams, Emily P.;Simon, M. Celeste
通讯作者：
Simon, M. Celeste
DOI：
10.1016/0027-5107(94)90300-x
发表时间：
1994-05-01
期刊：
MUTATION RESEARCH
影响因子：
--
作者：
ANDERSON, D;YU, TW;SCHMEZER, P
通讯作者：
SCHMEZER, P