A multichannel electrochemical platform for controlled hydrogen peroxide modulation in redox biology

A multichannel electrochemical platform for controlled hydrogen peroxide modulation in redox biology

Abstract

Reactive oxygen species (ROS) play a fundamental role in redox biology, impacting cellular signaling, oxidative stress responses, and disease progression. However, studying their effects in vitro remains challenging due to the difficulty of precisely controlling ROS levels over time. Traditional approaches rely on manual additions of H2O2, which lead to rapid concentration fluctuations and poor reproducibility. Here, we introduce a 96-well plate-compatible electrochemical platform that enables continuous, real-time modulation of H2O2 levels in cell culture by leveraging the oxygen reduction reaction. By precisely adjusting the applied electrical input, we generate defined amounts of H2O2 directly in solution, allowing dynamic and sustained ROS exposure without the need for frequent media changes or external chemical additions. To demonstrate its biological relevance, we tested the system on two cancer cell lines with differing resistance to oxidative stress. Controlled H2O2 generation induced dose-dependent cell death in both melanoma (A375) and glioblastoma (U87) cells, with U87 cells displaying greater resilience, consistent with previously reported findings. The addition of catalase fully prevented cytotoxicity, confirming that the observed effects were H2O2-specific. This variability in sensitivity highlights the potential of our platform for studying cell-type-dependent oxidative stress responses in a controlled manner. Beyond cancer models, this electrochemical approach provides a versatile tool for studying redox regulation in vitro with precise temporal and dosage control. It allows researchers to mimic physiological or pathological ROS exposure with improved reproducibility over traditional methods. Potential applications extend to oxidative stress research in neurodegeneration, inflammation, and redox-based therapeutics, where tightly regulated ROS levels are essential for understanding cellular responses.

Reactive oxygen species (ROS) play a fundamental role in redox biology, impacting cellular signaling, oxidative stress responses, and disease progression. However, studying their effects in vitro remains challenging due to the difficulty of precisely controlling ROS levels over time. Traditional approaches rely on manual additions of H2O2, which lead to rapid concentration fluctuations and poor reproducibility. Here, we introduce a 96-well plate-compatible electrochemical platform that enables continuous, real-time modulation of H2O2 levels in cell culture by leveraging the oxygen reduction reaction. By precisely adjusting the applied electrical input, we generate defined amounts of H2O2 directly in solution, allowing dynamic and sustained ROS exposure without the need for frequent media changes or external chemical additions. To demonstrate its biological relevance, we tested the system on two cancer cell lines with differing resistance to oxidative stress. Controlled H2O2 generation induced dose-dependent cell death in both melanoma (A375) and glioblastoma (U87) cells, with U87 cells displaying greater resilience, consistent with previously reported findings. The addition of catalase fully prevented cytotoxicity, confirming that the observed effects were H2O2-specific. This variability in sensitivity highlights the potential of our platform for studying cell-type-dependent oxidative stress responses in a controlled manner. Beyond cancer models, this electrochemical approach provides a versatile tool for studying redox regulation in vitro with precise temporal and dosage control. It allows researchers to mimic physiological or pathological ROS exposure with improved reproducibility over traditional methods. Potential applications extend to oxidative stress research in neurodegeneration, inflammation, and redox-based therapeutics, where tightly regulated ROS levels are essential for understanding cellular responses.

JAKEŠOVÁ, M.; EHLICH, J.; ERSCHEN, S.; NEMESKERI, L.; HANDL, V.; SCHINDL, R.; WALDHERR, L.; GLOWACKI, E.

01.06.2025

ELSEVIER SCIENCE INC

NEW YORK

FREE RADICAL BIOLOGY AND MEDICINE

233

United States of America

2

https://www.sciencedirect.com/science/article/pii/S0891584925003971?via%3Dihub