Mucus-coated, magnetically-propelled fecal surrogate to mimic fecal shear forces on colonic epithelium

Alan S Wang; Cecilia Villegas-Novoa; Yuli Wang; Christopher E Sims; Nancy L Allbritton

doi:10.1016/j.biomaterials.2024.122577

Mucus-coated, magnetically-propelled fecal surrogate to mimic fecal shear forces on colonic epithelium

Biomaterials. 2024 Apr 23:309:122577. doi: 10.1016/j.biomaterials.2024.122577. Online ahead of print.

Authors

Alan S Wang¹, Cecilia Villegas-Novoa², Yuli Wang², Christopher E Sims³, Nancy L Allbritton⁴

Affiliations

¹ Department of Bioengineering, University of Washington, Seattle, WA, 98195, USA; Division of Biological Sciences, University of California San Diego, La Jolla, CA, 92093, USA.
² Department of Bioengineering, University of Washington, Seattle, WA, 98195, USA.
³ Department of Bioengineering, University of Washington, Seattle, WA, 98195, USA; Department of Medicine, University of Washington, Seattle, WA, 98195, USA.
⁴ Department of Bioengineering, University of Washington, Seattle, WA, 98195, USA. Electronic address: nlallbri@uw.edu.

PMID: 38677221
DOI: 10.1016/j.biomaterials.2024.122577

Abstract

The relationship between the mechanical forces associated with bowel movement and colonic mucosal physiology is understudied. This is partly due to the limited availability of physiologically relevant fecal models that can exert these mechanical stimuli in in vitro colon models in a simple-to-implement manner. In this report, we created a mucus-coated fecal surrogate that was magnetically propelled to produce a controllable sweeping mechanical stimulation on primary intestinal epithelial cell monolayers. The mucus layer was derived from purified porcine stomach mucins, which were first modified with reactive vinyl sulfone (VS) groups followed by reaction with a thiol crosslinker (PEG-4SH) via a Michael addition click reaction. Formation of mucus hydrogel network was achieved at the optimal mixing ratio at 2.5 % w/v mucin-VS and 0.5 % w/v PEG-4SH. The artificial mucus layer possessed similar properties as the native mucus in terms of its storage modulus (66 Pa) and barrier function (resistance to penetration by 1-μm microbeads). This soft, but mechanically resilient mucus layer was covalently linked to a stiff fecal hydrogel surrogate (based on agarose and magnetic particles, with a storage modulus of 4600 Pa). The covalent bonding between the mucus and agarose ensured its stability in the subsequent fecal sliding movement when tested at travel distances as long as 203 m. The mucus layer served as a lubricant and protected epithelial cells from the moving fecal surrogate over a 1 h time without cell damage. To demonstrate its utility, this mucus-coated fecal surrogate was used to mechanically stimulate a fully differentiated, in vitro primary colon epithelium, and the physiological stimulated response of mucin-2 (MUC2), interleukin-8 (IL-8) and serotonin (5HT) secretion was quantified. Compared with a static control, mechanical stimulation caused a significant increase in MUC2 secretion into luminal compartment (6.4 × ), a small but significant increase in IL-8 secretion (2.5 × and 3.5 × , at both luminal and basal compartments, respectively), and no detectable alteration in 5HT secretion. This mucus-coated fecal surrogate is expected to be useful in in vitro colon organ-on-chips and microphysiological systems to facilitate the investigation of feces-induced mechanical stimulation on intestinal physiology and pathology.

Keywords: Colon; Fecal surrogate; Hydrogel; Magnetic; Michael addition; Mucus; Primary cells; Vinyl sulfone.