The Importance of Multifrequency Impedance Sensing of Endothelial Barrier Formation Using ECIS Technology for the Generation of a Strong and Durable Paracellular Barrier

Abstract

In this paper, we demonstrate the application of electrical cell-substrate impedance sensing (ECIS) technology for measuring differences in the formation of a strong and durable endothelial barrier model. In addition, we highlight the capacity of ECIS technology to model the parameters of the physical barrier associated with (I) the paracellular space (referred to as Rb) and (II) the basal adhesion of the endothelial cells (α, alpha). Physiologically, both parameters are very important for the correct formation of endothelial barriers. ECIS technology is the only commercially available technology that can measure and model these parameters independently of each other, which is important in the context of ascertaining whether a change in overall barrier resistance (R) occurs because of molecular changes in the paracellular junctional molecules or changes in the basal adhesion molecules. Finally, we show that the temporal changes observed in the paracellular Rb can be associated with changes in specific junctional proteins (CD144, ZO-1, and catenins), which have major roles in governing the overall strength of the junctional communication between neighbouring endothelial cells.

Interdigitated aluminium and titanium sensors for assessing epithelial barrier functionality by electric cell-substrate impedance spectroscopy (ECIS)

“…The increase in R cleft may be attributed to the excellent biocompatibility of titan which fosters strong adhesion between HeLa cells and the underlying titan electrodes (Saini et al 2015). The cell layer capacitance exhibited initial high values followed by a stabilization at around 1 μFcm −2 , which corresponds with reported values for cell membrane capacitance in other studies (Bagnaninchi and Drummond 2011;Robilliard et al 2018).…”

“…4 Equivalent electric circuit of the cell layer. To determine electrical parameters describing the cell layer, an equivalent electric circuit was applied according to Robilliard et al (2018). The model consists of the ohmic resistances R bulk (describing the resistance of the electrode traces and the resistance of the culture medium), R cell-layer (describing the resistance of the cell layer), and R cleft (describing the resistance of the subcellular cleft between the basolateral side of the cells and the electrode).…”

“…The first model is based on the work by Robilliard et al (2018) and consists of the ohmic resistances R bulk (representing the combined resistance of the cell culture medium, electrode traces, and measurement wiring), R cleft (resistance of the sub-cellular space underneath the cell layer), and R cell-layer (resistance of the cell layer). The capacitive elements of the EEC were C interface (interface capacitance) and C cell-layer (capacitance of the cell layer) (see Fig.…”

“…The EEC for modelling the cell layer is shown in Fig. 4 and is based on the work by Robilliard et al (2018). For HeLa cells, the resistance R cell-layer increased with cultivation time (see Fig.…”

Brain metastases-derived extracellular vesicles induce binding and aggregation of low-density lipoprotein

“…2 e). D-mannitol was used as a positive control, as it is known to damage the structure of specialized endothelial barriers in vitro [ 34 ]. Exposure of HBMEC monolayers to Reg-EVs and Br-EVs caused a similar increase in both dextran (61% increase for Reg-EVs and 69% increase for Br-EVs) and albumin (47% increase for Reg-EVs and 32% increase for Br-EVs) leakage (Fig.…”