Исследование проницаемости эпителиального слоя клеток TEER методом

Abstract

The study investigated the electrical characteristics of the Caco-2 cellular layer grown on a cellulose acetate membrane using impedance spectroscopy and cyclic voltammetry methods. The effect of ethanol solutions of varying concentrations on the electrophysical properties of the cellular layer was studied. During measurements of the layer characteristics by the cyclic voltammetry method under direct current, it was found that its resistance before ethanol treatment was 642 kOhm. Treatment with a 25% ethanol solution increased this value to 645 kOhm, while 50% and 95% solutions decreased it to 505 kOhm and 373 kOhm, respectively. During the study of the cellular layer's properties using impedance spectroscopy with alternating current, dependencies of the real and imaginary parts of the resistance on the ethanol concentration in the treatment solution were obtained. The obtained data were used to construct Nyquist diagrams. Their analysis allowed for the proposal of an equivalent circuit describing the process of current flow through the studied system. The equivalent circuit included parameters of paracellular transport, membrane resistance and capacitance, as well as elements accounting for electrode polarization. As a result, values of capacitance and membrane layer resistance were obtained. It was established that the membrane layer capacitance changes under the influence of ethanol. When treated with a 50% ethanol solution, capacitance increases, indicating degradation of the cell membrane and a reduction in its barrier function. Treatment with a 95% solution led to a decrease in capacitance, which can be attributed to the compression of the cell layer, reducing the capacitance contribution to the system's overall conductivity. The results demonstrate the importance of quantitative analysis of cellular electrical parameters for studying their response to chemical stress and confirm the potential of such approaches in biomedicine for creating sensors and «organ-on-a-chip» systems.