Quantitative Assessment of Ferroptosis in Cardiomyocytes Using Robust and Reliable Electrophysiological Biosensing.

Ferroptosis is an iron-dependent form of regulated cell death driven by dysregulated iron metabolism and subsequent lipid peroxidation, which contributes significantly to cardiovascular pathogenesis. While maintaining iron homeostasis is crucial for cardiac function, excessive iron accumulation initiates ferroptotic cell death, leading to cardiomyocyte damage. However, conventional biological methods lack the ability to dynamically and continuously monitor this process. To overcome this limitation, we developed a new biosensing platform centered on a custom 32-channel microelectrode array, which is designed for the non-invasive, label-free, and quantitative detection of electrophysiological signatures characteristic of cardiomyocyte ferroptosis. Using this platform, we successfully captured the drug concentration- and time-dependent electrophysiological alterations during Erastin-induced ferroptosis. Importantly, these detected electrophysiological changes closely aligned with traditional biochemical assays, accurately reflecting the progression of cardiomyocyte ferroptosis. The platform further demonstrated high specificity by recording the rescue effects of ferrostatin-1. Overall, our findings establish the feasibility of visualizing cardiomyocyte ferroptosis through electrophysiological monitoring, enabling continuous tracking of the entire process. This integrated biosensing system provides not only a powerful tool for investigating ferroptosis mechanisms but also a promising platform for drug screening and studying ferroptosis-related cardiac pathologies.