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Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies

Skin barrier dysfunction is important in atopic dermatitis and can be secondary to inflammation. Observation of keratinocytes in culture may show intrinsic differences. TransEpithelial Electrical Resistance (TEER) measures epithelial permeability. We cultured normal and atopic keratinocytes and foun...

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Autores principales: Marsella, Rosanna, Ahrens, Kim, Wilkes, Rachel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9319359/
https://www.ncbi.nlm.nih.gov/pubmed/35878346
http://dx.doi.org/10.3390/vetsci9070329
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author Marsella, Rosanna
Ahrens, Kim
Wilkes, Rachel
author_facet Marsella, Rosanna
Ahrens, Kim
Wilkes, Rachel
author_sort Marsella, Rosanna
collection PubMed
description Skin barrier dysfunction is important in atopic dermatitis and can be secondary to inflammation. Observation of keratinocytes in culture may show intrinsic differences. TransEpithelial Electrical Resistance (TEER) measures epithelial permeability. We cultured normal and atopic keratinocytes and found that TEER of atopic keratinocytes was significantly lower (p < 0.0001) than that of normals. Atopic keratinocytes grew upwards, first creating isolated dome-like structures and later horizontally into a monolayer. At time of confluence (D0), atopic keratinocytes were more differentiated, with higher filaggrin gene expression than normals. No differences existed between groups for TJ proteins (claudin, occludin, and Zonula Occludens-1) on D0 and D6. On D6, claudin and occludin were higher than D0, in normal (p = 0.0296 and p = 0.0011) and atopic keratinocytes (p = 0.0348 and 0.0491). Immunofluorescent staining showed nuclear location of filaggrin on D0 and cytoplasmic on D6. ANOVA showed increased cell size from D0 to D6 in both groups (effect of time, p = 0.0076) but no differences between groups. Significant subject effect (p = 0.0022) was found, indicating that cell size was subject-dependent but not disease-dependent. No difference for continuity for TJ protein existed between groups. These observations suggest that decreased TEER in atopics is not linked to TJ differences but is possibly linked to different growth behavior.
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spelling pubmed-93193592022-07-27 Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies Marsella, Rosanna Ahrens, Kim Wilkes, Rachel Vet Sci Communication Skin barrier dysfunction is important in atopic dermatitis and can be secondary to inflammation. Observation of keratinocytes in culture may show intrinsic differences. TransEpithelial Electrical Resistance (TEER) measures epithelial permeability. We cultured normal and atopic keratinocytes and found that TEER of atopic keratinocytes was significantly lower (p < 0.0001) than that of normals. Atopic keratinocytes grew upwards, first creating isolated dome-like structures and later horizontally into a monolayer. At time of confluence (D0), atopic keratinocytes were more differentiated, with higher filaggrin gene expression than normals. No differences existed between groups for TJ proteins (claudin, occludin, and Zonula Occludens-1) on D0 and D6. On D6, claudin and occludin were higher than D0, in normal (p = 0.0296 and p = 0.0011) and atopic keratinocytes (p = 0.0348 and 0.0491). Immunofluorescent staining showed nuclear location of filaggrin on D0 and cytoplasmic on D6. ANOVA showed increased cell size from D0 to D6 in both groups (effect of time, p = 0.0076) but no differences between groups. Significant subject effect (p = 0.0022) was found, indicating that cell size was subject-dependent but not disease-dependent. No difference for continuity for TJ protein existed between groups. These observations suggest that decreased TEER in atopics is not linked to TJ differences but is possibly linked to different growth behavior. MDPI 2022-06-30 /pmc/articles/PMC9319359/ /pubmed/35878346 http://dx.doi.org/10.3390/vetsci9070329 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Marsella, Rosanna
Ahrens, Kim
Wilkes, Rachel
Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies
title Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies
title_full Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies
title_fullStr Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies
title_full_unstemmed Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies
title_short Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies
title_sort differences in behavior between normal and atopic keratinocytes in culture: pilot studies
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9319359/
https://www.ncbi.nlm.nih.gov/pubmed/35878346
http://dx.doi.org/10.3390/vetsci9070329
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