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无标记光学生物传感器实时监测外源性毒物细胞毒性:草甘膦原理研究验证。

Label-free optical biosensor for real-time monitoring the cytotoxicity of xenobiotics: A proof of principle study on glyphosate.

机构信息

Nanobiosensorics Momentum Group, Institute of Technical Physics and Materials Science, Centre for Energy Research, Hungarian Academy of Sciences, Konkoly-Thege M. út 29-33, H-1120 Budapest, Hungary; Subdoctoral School of Molecular and Nanotechnologies, Chemical Engineering and Material Science Doctoral School, University of Pannonia, Egyetem u.10, H-8200 Veszprém, Hungary.

Agro-Environmental Research Institute, National Agricultural Research and Innovation Centre, Herman Ottó u. 15, H-1022 Budapest, Hungary.

出版信息

J Hazard Mater. 2018 Jun 5;351:80-89. doi: 10.1016/j.jhazmat.2018.02.045. Epub 2018 Feb 23.

Abstract

Rapid and inexpensive biosensor technologies allowing real-time analysis of biomolecular and cellular events have become the basis of next-generation cell-based screening techniques. Our work opens up novel opportunities in the application of the high-throughput label-free Epic BenchTop optical biosensor in cell toxicity studies. The Epic technology records integrated cellular responses about changes in cell morphology and dynamic mass redistribution of cellular contents at the 100-150 nm layer above the sensor surface. The aim of the present study was to apply this novel technology to identify the effect of the herbicide Roundup Classic, its co-formulant polyethoxylated tallow amine (POEA), and its active ingredient glyphosate, on MC3T3-E1 cells adhered on the biosensor surface. The half maximal inhibitory concentrations of Roundup Classic, POEA and glyphosate upon 1 h of exposure were found to be 0.024%, 0.021% and 0.163% in serum-containing medium and 0.028%, 0.019% and 0.538% in serum-free conditions, respectively (at concentrations equivalent to the diluted Roundup solution). These results showed a good correlation with parallel end-point assays, demonstrating the outstanding utility of the Epic technique in cytotoxicity screening, allowing not only high-throughput, real-time detection, but also reduced assay run time and cytotoxicity assessment at end-points far before cell death would occur.

摘要

快速且廉价的生物传感器技术能够实时分析生物分子和细胞事件,已成为下一代基于细胞的筛选技术的基础。我们的工作为高通量无标记 Epic BenchTop 光学生物传感器在细胞毒性研究中的应用开辟了新的机会。Epic 技术记录了细胞形态变化和细胞内容物动态质量重分布的综合细胞反应,其深度在传感器表面上方 100-150nm 处。本研究旨在应用这项新技术来识别除草剂 Roundup Classic、其共溶剂聚氧乙基牛脂胺 (POEA) 和其有效成分草甘膦对附着在生物传感器表面的 MC3T3-E1 细胞的影响。结果发现,在含血清培养基中,Roundup Classic、POEA 和草甘膦暴露 1 小时的半最大抑制浓度分别为 0.024%、0.021%和 0.163%,在无血清条件下分别为 0.028%、0.019%和 0.538%(相当于稀释后的 Roundup 溶液浓度)。这些结果与平行终点测定法相关性良好,表明 Epic 技术在细胞毒性筛选中具有出色的实用性,不仅能够进行高通量、实时检测,还能减少测定时间,并在远远早于细胞死亡发生之前的终点评估细胞毒性。

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