Quantifying Barrier Integrity in Organ-on-Chip Platforms
The EVOM™ Chip platform delivers high fidelity, real time TEER measurements that resolve barrier maturation and inflammatory disruption across endothelial and BBB models, enabling standardized, scalable assessment of barrier function in microphysiological systems.
Abstract
Real-time, quantitative assessment of barrier integrity is foundational for modeling neuro-vascular physiology and blood-brain barrier (BBB) function in vitro. In this study, the EVOM™ Chip platform was used to monitor trans-endothelial electrical resistance (TEER) in both a vascular endothelium and primary BBB tri-culture models. TEER measurements demonstrated robust barrier formation over time and detected significant barrier disruption following exposure to the pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-α). These findings establish the EVOM™ Chip platform as a highly sensitive, standardized, and scalable solution for resolving subtle barrier dynamics and inflammatory responses in advanced cell culture models.
Introduction
Barrier-forming tissues such as vascular endothelium and the BBB tightly regulate molecular exchange and tissue homeostasis, making their integrity a central readout in disease modeling, drug discovery, and toxicity testing. TEER is a widely adopted, non-invasive method for quantifying barrier formation and detecting barrier disruption. The EVOM™ Chip platform integrates embedded electrodes into microphysiological system formats to deliver sensitive, reproducible TEER measurements, eliminating manual variability and enabling continuous, hands-free monitoring. EVOM™ Chip delivers real-time, high-fidelity insights into cellular barrier dynamics while establishing a standardized approach for TEER measurement.
This application note demonstrates how the EVOM™ Chip platform is used to:
- Monitor endothelial barrier maturation in human umbilical vascular endothelial cell (HUVEC) monocultures through hands-free, multiplexed, on-chip TEER measurement.
- Evaluate BBB development in a primary tri-culture model on the SynBBB platform.
- Quantify cytokine-induced inflammatory barrier disruption using TEER as an indicator of functional disruption
Materials and Methods
HUVECs were cultured on EVOM™ Chip devices and monitored for barrier formation over 48 hours. A primary BBB model consisting of endothelial cells, astrocytes, and pericytes was established and cultured for 72-hours to promote barrier development and maturation. Following barrier establishment, cultures were treated with TNF-α (10 ng/mL) to induce barrier disruption. Barrier integrity was assessed by quantifying TEER on the EVOM™ Chip platform by measuring electrical resistance across the cellular barrier.
Statistical Analysis
Data are presented as mean ± SEM. HUVEC monoculture, n = 7; SynBBB tri-culture, n = 15. A one-way ANOVA with multiple comparisons was used to evaluate statistical significance. A significance threshold of p < 0.05 was applied. Significance levels: *** p < 0.001; **** p < 0.0001.
Results
A. EVOM™ Chip Quantifies HUVEC Barrier Formation and Cytokine-Mediated Disruption. Baseline resistance values averaged approximately 15 kΩ. Following 48 -hours of culture, TEER increased significantly to approximately 36 kΩ, indicating successful formation of a tight endothelial barrier. Exposure to TNF-α (10 ng/mL) for 4 -hours reduced resistance to approximately 29–30 kΩ, demonstrating a measurable loss of barrier integrity.
- TEER values more than doubled during monolayer maturation, reflecting rapid establishment of a high integrity endothelial barrier.
- TNF α exposure produced a rapid and quantifiable drop in TEER, demonstrating the platform’s ability to resolve acute inflammatory barrier breakdown.
- Statistically significant differences were observed between all experimental groups, despite a small sample size (n=7, ***p < 0.001, ****p < 0.0001).
Fig. 1 HUVEC Monoculture on EVOM™ Chip Platform
B. EVOM™ Chip Quantifies Complex BBB Formation and Injury. The SynBBB platform was used to grow primary tri-culture BBB cells and exhibited progressive barrier maturation, with TEER increasing from approximately 14–15 kΩ at baseline to approximately 25–26 kΩ after 72- -hours of culture. Following TNF-α exposure, resistance decreased to approximately 21–22 kΩ, indicating disruption of BBB integrity.
- The primary tri-culture model exhibited physiologically relevant BBB strengthening over 72-hours, consistent with coordinated endothelial–astrocyte–pericyte interactions.
- Cytokine challenge induced a clear reduction in TEER, confirming the model’s responsiveness to inflammatory stress and the platform’s sensitivity to subtle BBB perturbation.
- Statistically significant differences were observed between all timepoints, (n=15, ***p < 0.001, ****p < 0.0001).
Discussion
Accurate and quantitative measurement of barrier integrity is critical for studying vascular biology and modeling neurovascular disease mechanisms. Integrating TEER directly into organ on chip systems through WPI’s embedded electrode architecture provides a major advantage over traditional handheld probes, which require manual positioning, introduce operator variability, and disrupt culture conditions during each measurement. Embedded electrodes remain fixed within the device, enabling continuous, hands-free, non-invasive monitoring with consistent electrode geometry and superior signal stability, critical for resolving subtle changes in barrier function. In both endothelial monocultures and tri-cultures, this approach captured clear TEER increases during barrier maturation and rapid declines following inflammatory challenge, demonstrating the importance of incorporating TEER as a real time functional readout. Embedding TEER measurement directly into the culture platform ensures higher data quality, improved reproducibility, and more accurate interpretation of barrier dynamics in response to biological or pharmacological stimuli.
Here, we demonstrate the EVOM™ Chip can sensitively detect TEER increases reflecting barrier maturation in both monoculture and tri-culture cell models on SynBBB microfluidic chips, as well as quantify the effect of cytokines in inducing an inflammatory response and damaging the BBB. In both models, decreased TEER was observed following TNF-α treatment, consistent with cytokine-mediated disruption of tight junction structure and increased permeability of the barrier.
These results demonstrate that the EVOM™ Chip platform provides a sensitive, reproducible, and non-invasive method for monitoring barrier dynamics across a range of in vitro models. This technology establishes a foundation for standardized TEER based readouts across organ specific barrier models, enabling applications in drug screening [including high-throughput screening with the
These findings confirm that the EVOM™ Chip platform can sensitively detect both barrier formation and cytokine-induced barrier dysfunction in endothelial monocultures and BBB tri-cultures.
