Automated microfluidic-based polyurethane/lactate oxidase-chitosan/m-phenylenediamine biosensing platform for lactate monitoring in breast cancer cell culture

IF 5.7 2区化学 Q1 CHEMISTRY, ANALYTICAL

Analytica Chimica Acta Pub Date : 2025-04-10 DOI:10.1016/j.aca.2025.344049

Hnin Thandar Lwin , Kewarin Phonklam , Thanaporn Khunpitak , Pasarat Khongkow , Tonghathai Phairatana

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引用次数: 0

Abstract

Background

Lactate is a significant indicator of cancer characteristics. Monitoring lactate dynamics in cell metabolism provides crucial insights into cellular events, essential for developing novel therapeutic strategies, including drug screening. However, conventional methods for lactate detection in cell culture are typically time-consuming, labor-intensive, require large sample volumes, and offer only endpoint analysis.

Results

We introduce an automated microfluidic-based amperometric biosensor system for real-time lactate monitoring during a cell-culture experiment. An in-house combined needle microelectrode was modified with polyurethane/lactate oxidase-chitosan/m-phenylenediamine. The polyurethane layer was optimized to extend the sensor's linearity. Additionally, a computer-controlled microfluidic platform was developed to enable tracking of biosensor performance, ensuring precision in fluid delivery and improving the reliability of the monitoring process. The biosensor performances were monitored over time, demonstrating a sensitivity of 95.50 ± 6.30 nA/mM mm² with a detection limit of 82.60 ± 0.07 μM, and a wide linear range up to 6.0 mM. The lactate biosensor exhibited good selectivity for other potential molecules, while maintaining stability over 18 h during continuous monitoring. To assess its application in paclitaxel anti-cancer drug testing, we connected the analysis platform to a culture flask of MDA-MB-231 breast cancer cells and delivered the cell culture media samples to the analysis platform through a microdialysis probe.

Significance

The results revealed that the released extracellular lactate concentrations in the media were positively correlated with the CCK-8 (Cell Counting Kit-8) cell viability test results. Our lactate monitoring system shows significant potential as a practical tool for drug screening and for monitoring metabolic processes across various fields.

Abstract Image

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基于微流体的自动化聚氨酯/乳酸氧化酶-壳聚糖/间苯二胺生物传感平台用于乳腺癌细胞培养中乳酸监测

乳酸是癌症特征的重要指标。监测细胞代谢中的乳酸动态提供了对细胞事件的重要见解，对于开发包括药物筛选在内的新治疗策略至关重要。然而，在细胞培养中检测乳酸的传统方法通常耗时，劳动密集，需要大量的样本量，并且只提供终点分析。结果我们介绍了一种基于微流体的自动化安培生物传感器系统，用于在细胞培养实验中实时监测乳酸浓度。采用聚氨酯/乳酸氧化酶-壳聚糖/间苯二胺对室内组合针微电极进行了改性。优化了聚氨酯层以延长传感器的线性度。此外，开发了一个计算机控制的微流控平台，以跟踪生物传感器的性能，确保流体输送的精度，提高监测过程的可靠性。该传感器的灵敏度为95.50±6.30 nA/mM mm2，检出限为82.60±0.07 μM，线性范围为6.0 mM，对其他潜在分子具有良好的选择性，在连续监测18小时内保持稳定。为了评估其在紫杉醇抗癌药物测试中的应用，我们将分析平台与MDA-MB-231乳腺癌细胞培养瓶连接，并通过微透析探针将细胞培养基样品输送到分析平台。结果显示，培养基中释放的胞外乳酸浓度与CCK-8 （Cell Counting Kit-8）细胞活力测试结果呈正相关。我们的乳酸监测系统显示出作为药物筛选和监测各个领域代谢过程的实用工具的巨大潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Analytica Chimica Acta 化学-分析化学

CiteScore

10.40

自引率

6.50%

发文量

1081

审稿时长

38 days

期刊介绍： Analytica Chimica Acta has an open access mirror journal Analytica Chimica Acta: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. Analytica Chimica Acta provides a forum for the rapid publication of original research, and critical, comprehensive reviews dealing with all aspects of fundamental and applied modern analytical chemistry. The journal welcomes the submission of research papers which report studies concerning the development of new and significant analytical methodologies. In determining the suitability of submitted articles for publication, particular scrutiny will be placed on the degree of novelty and impact of the research and the extent to which it adds to the existing body of knowledge in analytical chemistry.