Broadband electrical impedance as a novel characterization of oxidative stress in single L6 skeletal muscle cells
Broadband electrical impedance as a novel characterization of oxidative stress in single L6 skeletal muscle cells
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DOI：
10.1016/j.aca.2021.338678
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发表时间：
2021-06-02
影响因子：
6.2
通讯作者：
Cheng, Xuanhong
中科院分区：
文献类型：
--
作者：
Ferguson, Caroline;Pini, Niccolo;Cheng, Xuanhong
氧化应激（OS）是细胞毒性的主要原因之一，与许多人类生理病理条件有关。特别是，肌痛性脑脊髓炎/慢性疲劳综合征（ME/CFS）引起的OS是衰弱的生活质量，而没有明确的生物标志物已被确定为诊断措施。最近，ME/CFS患者的外周血细胞的阻抗测量已被证明是一种有前途的方法来诊断疾病。受这项研究的启发，旨在直接询问肌肉细胞，我们研究了单个肌肉细胞的宽带测量是否可以区分正常和氧化应激细胞群。我们首先优化了一个方案，通过过氧化氢处理引入氧化应激培养大鼠L 6骨骼肌细胞。处理后的细胞进一步通过宽带阻抗谱的单细胞使用微流控芯片实验室系统。由此产生的细胞质介电常数和电导率的介电特性与正常培养的细胞在电学上不同。正常细胞的反射和透射系数Delta S11和Delta S21紧密聚集，并且在9 kHz至9 GHz的频率范围内与无细胞溶液的反射和透射系数非常相似。另一方面，氧化细胞的介电性质在GHz范围内具有宽的分布，与正常培养的细胞在正方向和负方向上都偏离。模拟结果指导我们的假设，即介电差异可能与离子的改变，而钙成像直接支持的贡献，钙通量的观察到的S参数的偏差。GHz频率下与氧化细胞相关的独特电特性为未来诊断氧化应激相关疾病（如ME/CFS）的技术开发提供了框架。（C）2021 Elsevier B. V.保留所有权利。
Oxidative stress (OS) is one of the leading causes of cytotoxicity and is linked to many human physio-pathological conditions. In particular, myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) induced by OS is debilitating to quality of life, while no clear biological markers have been identified for diagnostic measures. Recently, impedance measurements of peripheral blood cells of ME/CFS patients have been shown as a promising approach to diagnose the disease. Inspired by this study and aiming to interrogate muscle cells directly, we investigated if broadband measurements of single muscle cells could differentiate normal and oxidatively stressed cell populations. We first optimized a protocol through H2O2 treatment to introduce oxidative stress to cultured rat L6 skeletal muscle cells. The treated cells were further characterized through broadband impedance spectroscopy of single cells using a micro-fluidic lab-on-a-chip system. The resulting dielectric properties of cytoplasm permittivity and conductivity are electrically distinct from normally cultured cells. The reflection and transmission coefficients, Delta S11 and Delta S21, of the normal cells are tightly clustered and closely resemble those of the cell-free solution across the frequency range of 9 kHz to 9 GHz. On the other hand, dielectric properties of the oxidized cells have a wide distribution in the GHz range, deviating both in the positive and negative directions from the normally cultured cells. Simulation results guide our hypothesis that the dielectric differences could be linked to ion alterations, while calcium imaging directly supports the contribution of calcium flux to the observed deviation of S parameters. The unique electrical profile associated with oxidized cells in the GHz frequencies provide a framework for future development of technologies to diagnose oxidative-stress related diseases such as ME/CFS. (C)2021 Elsevier B.V. All rights reserved.