Issue 14, 2023

From mouth to gut: microfluidic in vitro simulation of human gastro-intestinal digestion and intestinal permeability

Abstract

Reproducible in vitro studies of bioaccessibility, intestinal absorption, and bioavailability are key to the successful development of novel food ingredients or drugs intended for oral administration. There is currently a lack of methods that offer the finesse required to study these parameters for valuable molecules typically found in small volumes – as is the case of nanomaterials, which are often used to carry and protect bioactives. Here, we describe a modular microfluidic-based platform for total simulation of the human gastro-intestinal tract. Digestion-chips and cell-based gut-chips were fabricated from PDMS by soft lithography. On-chip digestion was validated using a fluorescently labelled casein derivative, which followed typical Michaelis–Menten kinetics and showed temporal resolution and good agreement with well-established bench-top protocols. Irreversible inhibition of serine proteases using Pefabloc® SC and a 1 : 6 dilution was sufficient to mitigate the cytotoxicity of simulated digestion fluids. Caco-2/HT29-MTX co-cultures were grown on-chip under a continuous flow for 7 days to obtain a differentiated cell monolayer forming a 3D villi-like epithelium with clear tight junction formation, and with an apparent permeability (Papp) of Lucifer Yellow closely approximating values reported ex vivo (3.7 × 10−6 ± 1.4 × 10−6vs. 4.0 × 10−6 ± 2.2 × 10−6). Digesta from the digestion-chips were flowed through the gut-chip, demonstrating the capacity to study sample digestion and intestinal permeability in a single microfluidic platform holding great promise for use in pharmacokinetic studies.

Graphical abstract: From mouth to gut: microfluidic in vitro simulation of human gastro-intestinal digestion and intestinal permeability

Supplementary files

Article information

Article type
Paper
Submitted
28 Dec 2022
Accepted
06 May 2023
First published
09 May 2023
This article is Open Access
Creative Commons BY license

Analyst, 2023,148, 3193-3203

From mouth to gut: microfluidic in vitro simulation of human gastro-intestinal digestion and intestinal permeability

M. Xavier, P. M. Rodrigues, M. D. Neto, M. I. Guedes, V. Calero, L. Pastrana and C. Gonçalves, Analyst, 2023, 148, 3193 DOI: 10.1039/D2AN02088B

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements