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集成传感器的微生理系统研究脉冲电场的影响。

Microphysiological system with integrated sensors to study the effect of pulsed electric field.

机构信息

Laboratory of Bioelectrics, State Research Institute, Center for Physical Sciences and Technology, Sauletekio Ave. 3, 10257, Vilnius, Lithuania.

Nanostructured Materials and Sensors Laboratory, State Research Institute, Center for Physical Sciences and Technology, Sauletekio Ave. 3, 10257, Vilnius, Lithuania.

出版信息

Sci Rep. 2024 Aug 12;14(1):18713. doi: 10.1038/s41598-024-69693-w.

Abstract

This study focuses on the use of pulsed electric fields (PEF) in microfluidics for controlled cell studies. The commonly used material for soft lithography, polydimethylsiloxane (PDMS), does not fully ensure the necessary chemical and mechanical resistance in these systems. Integration of specific analytical measurement setups into microphysiological systems (MPS) are also challenging. We present an off-stoichiometry thiol-ene (OSTE)-based microchip, containing integrated electrodes for PEF and transepithelial electrical resistance (TEER) measurement and the equipment to monitor pH and oxygen concentration in situ. The effectiveness of the MPS was empirically demonstrated through PEF treatment of the C6 cells. The effects of PEF treatment on cell viability and permeability to the fluorescent dye DapI were tested in two modes: stop flow and continuous flow. The maximum permeability was achieved at 1.8 kV/cm with 16 pulses in stop flow mode and 64 pulses per cell in continuous flow mode, without compromising cell viability. Two integrated sensors detected changes in oxygen concentration before and after the PEF treatment, and the pH shifted towards alkalinity following PEF treatment. Therefore, our proof-of-concept technology serves as an MPS for PEF treatment of mammalian cells, enabling in situ physiological monitoring.

摘要

本研究专注于在微流控中使用脉冲电场 (PEF) 进行受控细胞研究。软光刻中常用的材料聚二甲基硅氧烷(PDMS)并不能完全确保这些系统所需的化学和机械稳定性。将特定的分析测量装置集成到微生理系统 (MPS) 中也具有挑战性。我们提出了一种基于非化学计量硫醇-烯 (OSTE) 的微芯片,其中包含用于 PEF 和跨上皮电阻 (TEER) 测量的集成电极,以及原位监测 pH 值和氧浓度的设备。通过对 C6 细胞进行 PEF 处理,经验证了 MPS 的有效性。通过两种模式:停止流动和连续流动,测试了 PEF 处理对细胞活力和对荧光染料 DapI 通透性的影响。在停止流动模式下,最大通透性在 1.8 kV/cm 时达到 16 个脉冲,在连续流动模式下每个细胞达到 64 个脉冲,而不会影响细胞活力。两个集成传感器在 PEF 处理前后检测到氧浓度的变化,并且 pH 值在 PEF 处理后向碱性偏移。因此,我们的概念验证技术可作为哺乳动物细胞 PEF 处理的 MPS,实现原位生理监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/641c/11319666/3a292a400be6/41598_2024_69693_Fig1_HTML.jpg

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