of water and mineral nutrients, [1,2] while in human body are the major building blocks of various organs, including, for example, skin, lungs, liver, kidney, and digestive track. The physiological function of the biological barriers is diverse among tissues and responds to the specific needs of each organ, including ion absorption, nutrients uptake, protection against toxins, and secretion of waste. [3,4] Under normal physiological conditions the transcellular and paracellular pathways are finely regulated by the cellular barriers. Tight-junction (TJ) proteins-being responsible of the intercellular sealingcontrol the paracellular fluxes, providing either fully impermeable barriers or permeable-selective functions. [5][6][7] The permeability of TJs in a cell barrier can be regulated by physiological cues, can be modulated by drugs, and can be altered by various biological events such as inflammation, gastrointestinal tract diseases, cancer metastases, leukocyte migration, and viral infections. For instance, disruption of epithelial and endothelial barriers is a key clinical data differentiating patients with high probability to develop severe COVID-19 symptoms including the escalation in respiratory deficiency, loss of viral containment, and a progression toward multiorgan dysfunction. [8] Analogously, blood-brain barrier disruption contributes to the severity of diverse neurological diseases, including stroke, epilepsy, Alzheimer's disease, and multiple sclerosis, among others. [9,10] More in general, monitoring cellular barriers with noninvasive and label-free methods is relevant for in vitro studies, [11] for the development of organ-on-chip models, [12] for studying a disease progression, and for drug testing and drug targeting, also promoting the replacement for animal testing in toxicological profiling. [13] So far, the integrity of cellular barriers has been addressed with transepithelial electrical resistance (TEER) measurements. TEER is an in vitro measurement technique based on two electrodes placed on each side of a cell layer. Upon the application of a direct or alternate current, the ionic impedance of the barrier layer is measured. [14] TEER direct current method is easy to perform and the measurement provides a resistance value that depends on both the cell status and the electrode positions. The positioning of the electrodes is performed manually, and this Cellular barriers control fundamental physiological functions in animals and plants. Accurate detection of barrier dysfunction requires real-time monitoring. Organic electrochemical transistors are a promising bioelectronic platform to monitoring cellular barriers. However, current approaches are not ideally suited for direct and real-time measurements: they require offline model-based data analysis or slow measurement operation to achieve equilibrium conditions. Herein, dynamic-mode current-driven organic electrochemical transistors are proposed for direct real-time monitoring of cellular barrier functionality. In contrast to current approaches...