Innovative platforms for quantitative in vitro characterization of cardiac electro-contractile activity

Cardiovascular diseases remain the leading cause of mortality worldwide, while cardiotoxic side effects continue to limit the safety and efficacy of many pharmacological therapies. Advanced experimental methods capable of capturing early and subtle functional alterations of cardiac tissue are therefore essential for improving pre-clinical investigation. In this context, in-vitro cardiac models represent powerful and versatile tools, providing quantitative, reproducible, and physiologically relevant assessment of both electrophysiological and contractile cardiac function. However, most existing platforms investigate these two aspects separately or provide only qualitative readouts averaged over large populations, limiting their translational relevance for disease modeling and drug screening. This PhD work focuses on the development and validation of multimodal experimental platforms for high throughput, integrated, and quantitative investigation of electro-contractile-mechanical properties in in-vitro cardiac models, with the aim of identifying functional biomarkers sensitive to pathological conditions and pharmacological interventions. In the first part of the work, a combined Atomic Force Microscopy–Microelectrode Array (AFM-MEA) platform was optimized to enable synchronized, single-cell-level measurements of extracellular field potentials, contractile behavior, and mechanical properties in a two-dimensional cardiac model. This system was employed to study doxorubicin-induced cardiotoxicity and to evaluate the cardioprotective potential of extracellular vesicles derived from human amniotic fluid stem cells (hAFS-EVs). The results reveal that doxorubicin induces marked alterations in both electrical and contractile parameters, while pre-treatment with extracellular vesicles partially mitigates these effects, highlighting their cardioprotective potential. In the second part, to overcome the low-throughput limitation of the AFM-MEA setup, a novel hybrid platform integrating an active Thin-Film Transistor Microelectrode Array (TFT-MEA) with a flexible Micro-Pillar Array (MPA) was developed. Preliminary results demonstrated the platform’s potential to enable high-throughput, label-free recording of both electrical and contractile activity from the same cardiomyocyte population. Pharmacological validation with isoprenaline confirmed the platform’s sensitivity and reliability for multimodal functional analysis and drug screening applications. Overall, this work proposes innovative experimental tools for the quantitative and integrated study of cardiac electro-contractile coupling in-vitro, either at single-cell level with low-throughput or at the cell population level with high-throughput. These approaches offer new opportunities for cardiotoxicity assessment and drug screening, and represent a step toward more predictive and safer preclinical evaluation of therapeutic strategies.