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Comparison of 2D, 3D In Vitro, and Ex Vivo Platforms for Modeling the Rat Small Intestine

Title: Comparison of 2D, 3D In Vitro, and Ex Vivo Platforms for Modeling the Rat Small Intestine
Authors: Shani Elias-Kirma; Reece McCoy; Douglas van Niekerk; Verena Stoeger; Sophie Oldroyd; Emma Sumner; Achilleas Savva; Róisín M. Owens
Source: Bioengineering ; Volume 13 ; Issue 3 ; Pages: 349
Publisher Information: Multidisciplinary Digital Publishing Institute
Publication Year: 2026
Collection: MDPI Open Access Publishing
Subject Terms: 3D in vitro; bioelectronics; PEDOT:PSS scaffold; rat; small intestine barrier
Description: Physiologically relevant in vitro intestinal models are essential for studying key physiological processes, including barrier function, drug screening and gut-microbiota interactions. However, conventional 2D culture systems often fail to recapitulate structural and functional complexity. Here, we aimed to validate a 3D bioelectronic transmembrane platform, previously used for monitoring human intestinal epithelium and vascular endothelium, for modeling the rat small intestinal barrier in vitro. The device integrates a poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) scaffold supporting co-cultures of rat intestinal epithelial cells (IEC-6) and rat fibroblasts (208F), enabling real-time monitoring of barrier formation through electrical measurements using electrochemical impedance spectroscopy (EIS). Barrier formation was monitored over 21 days and exhibited a time-dependent increase in barrier resistance. The 3D platform was compared with traditional 2D insert-based cultures and ex vivo rat tissue using an Ethylene Glycol Tetraacetic Acid (EGTA)-induced calcium switch assay to evaluate barrier disruption and recovery. EGTA treatment and removal induced reversible barrier disruption in the 3D in vitro and ex vivo models, whereas 2D in vitro cultures showed limited recovery. These findings demonstrate that the 3D platform more faithfully recapitulates native tissue architecture and function, closely paralleling ex vivo responses. Our study highlights the importance of validating advanced 3D in vitro models and establishes this bioelectronic platform as a robust tool for drug screening, barrier studies, and preclinical gastrointestinal research.
Document Type: text
File Description: application/pdf
Language: English
Relation: Regenerative Engineering; https://dx.doi.org/10.3390/bioengineering13030349
DOI: 10.3390/bioengineering13030349
Availability: https://doi.org/10.3390/bioengineering13030349
Rights: https://creativecommons.org/licenses/by/4.0/
Accession Number: edsbas.904005C6
Database: BASE