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Therefore, there is a need to develop novel microfluidic models that mimic the intestinal biological and mechanical environments. These models can be used for drug discovery and disease modeling and have the potential to reduce reliance on animal models. The goal of this study was to develop a small intestine on a chip with both enterocyte (Caco\u20102) and goblet (HT29\u2010MTX) cells cocultured with <jats:italic>Lacticaseibacillus rhamnosus<\/jats:italic> biofilms, which is of one of several genera present in the small intestinal microbiota. <jats:italic>L. rhamnosus<\/jats:italic> was introduced following the establishment of the epithelial barrier. The shear stress within the device was kept in the lower physiological range (0.3\u2009mPa) to enable biofilm development over the in vitro epithelium. The epithelial barrier differentiated after 5 days of dynamic culture with cell polarity and permeability similar to the human small intestine. The presence of biofilms did not alter the barrier's permeability in dynamic conditions. Under fluid flow, the complete model remained viable and functional for more than 5 days, while the static model remained functional for only 1 day. The presence of biofilm increased the secretion of acidic and neutral mucins by the epithelial barrier. Furthermore, the small intestine on a chip also showed increased MUC2 production, which is a dominant gel\u2010forming mucin in the small intestine. This model builds on previous publications as it establishes a stable environment that closely mimics in vivo conditions and can be used to study intestinal physiology, food\u2010intestinal interactions, and drug development.<\/jats:p>","DOI":"10.1002\/bit.28989","type":"journal-article","created":{"date-parts":[[2025,4,8]],"date-time":"2025-04-08T09:37:16Z","timestamp":1744105036000},"page":"1816-1827","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Small Intestine on a Chip Demonstrates Physiologic Mucus Secretion in the Presence of <i>Lacticaseibacillus rhamnosus<\/i> Biofilm"],"prefix":"10.1002","volume":"122","author":[{"given":"Sanat Kumar","family":"Dash","sequence":"first","affiliation":[{"name":"Department of Biomedical Engineering Binghamton University Binghamton New York USA"},{"name":"Department of Biological Sciences Binghamton University Binghamton New York USA"},{"name":"Binghamton Biofilm Research Center Binghamton University Binghamton New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2873-7074","authenticated-orcid":false,"given":"Cla\u0301udia N. 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