<i>In vitro</i>assessment of epithelial electrical resistance in human esophageal and jejunal mucosae and in Caco-2 cell layers

Björkman, Eleonora; Casselbrant, Anna; Lundberg, Sören; Fändriks, Lars

doi:10.3109/00365521.2012.722677

Cited by 19 publications

(16 citation statements)

References 37 publications

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“…We confirmed the findings of previous studies showing that the non-eroded mucosa of patients with erosive esophagitis has an increased permeability to ions, as shown by a lower TEER [24,25]. In contrast to the ex vivo findings, in vivo mucosal impedance values in that and many other studies showed a remarkable downward gradient from healthy controls to NERD to EE [21,22,26,27,28].…”

Section: Discussionsupporting

confidence: 91%

Exploration of the Esophageal Mucosal Barrier in Non-Erosive Reflux Disease

Rinsma

Farré

Troost

et al. 2017

IJMS

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In the absence of visible mucosal damage, it is hypothesized that the esophageal mucosal barrier is functionally impaired in patients with non-erosive reflux disease (NERD). The aim of the present study was to perform an exploratory analysis of the mucosal barrier in NERD compared to erosive esophagitis (EE) and controls. A second aim was to explore TRPV1 gene transcription in relation to the mucosal barrier function and heartburn symptoms. In this prospective study, 10 NERD patients, 11 patients with active erosive esophagitis and 10 healthy volunteers were included. Biopsies from non-eroded mucosa were obtained for (1) ex vivo analyses (Ussing chamber) of transepithelial electrical resistance (TEER) and permeability (2) gene transcription of tight-junction proteins and transient receptor potential vanilloid subfamily member 1 (TRPV1). No differences in TEER or permeability were found between NERD and healthy volunteers, whereas TEER was lower in patients with erosive esophagitis. TRPV1 gene transcription was not significantly different between EE, NERD and controls. Conclusions: esophageal mucosal barrier function and TRPV1 transcription is not significantly altered in NERD patients. Future research is needed to explore other potential mechanisms that may account for the high symptom burden in these patients.

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Section: Discussionsupporting

confidence: 91%

Exploration of the Esophageal Mucosal Barrier in Non-Erosive Reflux Disease

Rinsma

Farré

Troost

et al. 2017

IJMS

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“…While we observed no difference in permeability between our groups ( Figure 1A), our low number of subjects coupled with the enormous heterogeneity observed in IBD prevents us from proving or disproving this hypothesis but the finding that P app of FD4 can be assessed consistently in multiple patient iPSC-derived lines in Transwell inserts confirms the technical feasibility of this approach and now permits an examination of this hypothesis by including a much larger number of subjects within each group. The basal P app of FD4 (0.57-0.92 × 10 −7 cm/s) compared favorably with human small intestine (0.5-2.1 × 10 −7 cm/s) recordings in Ussing chambers [33,34]. Additionally, our P app values along with our basal TEER values (404-615 Ω•cm 2 ; Figure 2B) also compared favorably to a recent study which reported a basal TEER of 420 Ω•cm 2 and P app value of 0.12 × 10 −6 cm/s using iPSC derived intestinal epithelial cell monolayers generated by CDX2 transduction [35].…”

Section: Discussionmentioning

confidence: 79%

Development of Physiologically Responsive Human iPSC-Derived Intestinal Epithelium to Study Barrier Dysfunction in IBD

Gleeson

Estrada

Yamashita

et al. 2020

IJMS

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In inflammatory bowel disease (IBD), the intestinal epithelium is characterized by increased permeability both in active disease and remission states. The genetic underpinnings of this increased intestinal permeability are largely unstudied, in part due to a lack of appropriate modelling systems. Our aim is to develop an in vitro model of intestinal permeability using induced pluripotent stem cell (iPSC)-derived human intestinal organoids (HIOs) and human colonic organoids (HCOs) to study barrier dysfunction. iPSCs were generated from healthy controls, adult onset IBD, and very early onset IBD (VEO-IBD) patients and differentiated into HIOs and HCOs. EpCAM+ selected cells were seeded onto Transwell inserts and barrier integrity studies were carried out in the presence or absence of pro-inflammatory cytokines TNFα and IFNγ. Quantitative real-time PCR (qRT-PCR), transmission electron microscopy (TEM), and immunofluorescence were used to determine altered tight and adherens junction protein expression or localization. Differentiation to HCO indicated an increased gene expression of CDX2, CD147, and CA2, and increased basal transepithelial electrical resistance compared to HIO. Permeability studies were carried out in HIO- and HCO-derived epithelium, and permeability of FD4 was significantly increased when exposed to TNFα and IFNγ. TEM and immunofluorescence imaging indicated a mislocalization of E-cadherin and ZO-1 in TNFα and IFNγ challenged organoids with a corresponding decrease in mRNA expression. Comparisons between HIO- and HCO-epithelium show a difference in gene expression, electrophysiology, and morphology: both are responsive to TNFα and IFNγ stimulation resulting in enhanced permeability, and changes in tight and adherens junction architecture. This data indicate that iPSC-derived HIOs and HCOs constitute an appropriate physiologically responsive model to study barrier dysfunction and the role of the epithelium in IBD and VEO-IBD.

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“…Since we were interested in the specific transepithelial resistance and not the total tissue resistance we tried to remove as much of the subepithelium as possible to minimize the influence on the TER. We are currently upgrading our systems to use square-pulse analysis for even more adequate assessment of the epithelial resistance 246 .…”

Section: Methodological Considerations Ussing Chambersmentioning

confidence: 99%

Role of mast cells and probiotics in the regulation of intestinal barrier function

Carlsson¹

2013

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About the coverThe cover displays immunofluorescence image of intestinal villus epithelium and mast cells. Nuclei are stained blue with DAPI and mast cell tryptase is stained green. The image was acquired on a Zeiss LSM 700.During the course of the research underlying this thesis, Anders Carlsson was enrolled in Forum GIMIICum, faa national research school in gastrointestinal infection and inflammation funded by the Swedish Research Council and based at Linköping University ABSTRACTThe intestinal mucosa is the largest contact area and one of the most important barriers to the outside environment. It is highly specialized in aiding us digest and absorb nutrients. It is daily exposed to several potentially dangerous substances and microorganisms, which if they were allowed to pass into the body, could give rise to diseases. Throughout the small intestine certain sites specialized in antigen sampling are found. These sites are named Peyer's patches and are lymphoid follicles. The epithelium covering the Peyer's patches is called follicle-associated epithelium and is specialized in antigen sampling and uptake. The special epithelium enables presentation of luminal antigen to immune cells in the underlying follicle. Persistent life stress and stressful life events affect the course of irritable bowel syndrome (IBS) and inflammatory bowel disease (IBD) through largely unknown mechanisms. Regulation of epithelial permeability to antigens is crucial for the balance between inflammation and immune-surveillance, and increased intestinal permeability has been shown in patients with ulcerative colitis and Crohns disease. Vasoactive intestinal polypeptide (VIP) and corticotropin-releasing factor have been implicated as important mediators of stress-induced abnormalities in intestinal mucosal functions in animal models. Both of these mediators have been reported to regulate bowel ion secretion in humans during stress and uptake of horseradish peroxidase in rodents. Probiotics have been shown to ameliorate the deleterious effects of stress on intestinal function, but mechanisms remain to be elucidated. The aim of this thesis was to elucidate whether mast cells play an important role in intestinal barrier function during stress and inflammation. Moreover, we wanted to determine whether probiotics can ameliorate the mucosal barrier integrity during stress and inflammation.To study the function of mast cells we conducted in vitro experiments on cell lines and ex vivo experiments in Ussing chambers on mouse, rat and human intestinal tissue. The Ussing chamber technique measures electrophysiological properties of the tissue and also gives the possibility to study transcellular and paracellular passage of markers and bacteria. Immunohistology and confocal microscopy have been used to identify mast cells and receptors of interest.Our results show that stress affects the follicle-associated epithelium barrier by mechanisms involving VIP and mast cells. These findings were corroborated by the localization of VIP receptors on mucosal ...

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In vitroassessment of epithelial electrical resistance in human esophageal and jejunal mucosae and in Caco-2 cell layers

Cited by 19 publications

References 37 publications

Exploration of the Esophageal Mucosal Barrier in Non-Erosive Reflux Disease

Exploration of the Esophageal Mucosal Barrier in Non-Erosive Reflux Disease

Development of Physiologically Responsive Human iPSC-Derived Intestinal Epithelium to Study Barrier Dysfunction in IBD

Role of mast cells and probiotics in the regulation of intestinal barrier function

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