Reconstructed human epidermis: A model to study the barrier function

The main function of the skin is to protect the body from the external environment. The barrier function of the skin is mainly provided by the stratum corneum, which consists of corneocytes bound with the corneodesmosomes and lamellar lipids. Skin barrier proteins like loricrin and filaggrin also contribute to the skin barrier function. In various skin diseases, skin barrier dysfunction is a common symptom, and skin irritants like detergents or surfactants could also perturb skin barrier function. Many efforts have been made to develop strategies to improve skin barrier function. Here, we investigated whether the microfluidized lysates of Lactobacillus rhamnosus (LR), one of the most widely used probiotic species for various health benefits, may improve the skin barrier function in a reconstructed human epidermis, Keraskin™. Application of LR lysate on Keraskin™ increased the expression of tight junction proteins; claudin 1 and occludin as determined by immunofluorescence analysis, and skin barrier proteins; loricrin and filaggrin as determined by immunohistochemistry and immunofluorescence analysis and qPCR. Also, the cytotoxicity of a skin irritant, sodium lauryl sulfate (SLS), was alleviated by the pretreatment of LR lysate. The skin barrier protective effects of LR lysate could be further demonstrated by the attenuation of SLS-enhanced dye-penetration. LR lysate also attenuated the destruction of desmosomes after SLS treatment. Collectively, we demonstrated that LR lysate has protective effects on the skin barrier, which could expand the utility of probiotics to skin-moisturization ingredients.

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Freezing Weakens the Barrier Function of Reconstructed Human Epidermis as Evidenced by Raman Spectroscopy and Percutaneous Permeation

Pharmaceutics ◽

10.3390/pharmaceutics12111041 ◽

2020 ◽

Vol 12 (11) ◽

pp. 1041

Author(s):

Yuri Dancik ◽

Hichem Kichou ◽

Christophe Eklouh-Molinier ◽

Martin Soucé ◽

Emilie Munnier ◽

...

Keyword(s):

Barrier Function ◽

Principal Component ◽

Human Epidermis ◽

Animal Tissues ◽

Reconstructed Human Epidermis ◽

Percutaneous Permeation ◽

Time Required ◽

Kinetics Of ◽

Permeation Kinetics

The development and characterization of reconstructed human epidermis (RHE) is an active area of R&D. RHE can replace animal tissues in pharmaceutical, toxicological and cosmetic sciences, yielding scientific and ethical advantages. RHEs remain costly, however, due to consumables and time required for their culture and a short shelf-life. Storing, i.e., freezing RHE could help reduce costs but to date, little is known on the effects of freezing on the barrier function of RHE. We studied such effects using commercial EpiSkin™ RHE stored at −20, −80 and −150 °C for 1 and 10 weeks. We acquired intrinsic Raman spectra in the stratum corneum (SC) of the RHEs as well as spectra obtained following topical application of resorcinol in an aqueous solution. In parallel, we quantified the effects of freezing on the permeation kinetics of resorcinol from time-dependent permeation experiments. Principal component analyses discriminated the intrinsic SC spectra and the spectra of resorcinol-containing RHEs, in each case on the basis of the freezing conditions. Permeation of resorcinol through the frozen RHE increased 3- to 6-fold compared to fresh RHE, with the strongest effect obtained from freezing at −20 °C for 10 weeks. Due to the extensive optimization and standardization of EpiSkin™ RHE, the effects observed in our work may be expected to be more pronounced with other RHEs.

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Reconstructed Human Epidermis: A Model to Study in vitro the Barrier Function of the Skin

Skin Pharmacology and Physiology ◽

10.1159/000211017 ◽

1992 ◽

Vol 5 (1) ◽

pp. 49-56 ◽

Cited By ~ 33

Author(s):

M. Régnier ◽

D. Caron ◽

U. Reichert ◽

H. Schaefer

Keyword(s):

Barrier Function ◽

Human Epidermis ◽

Reconstructed Human Epidermis

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Treatment with DHA Improves Epidermal Eeratinocyte Differentiation and Ameliorates Inflammation in Human Keratinocytes and Reconstructed Human Epidermis Models

10.20944/preprints201908.0178.v1 ◽

2019 ◽

Author(s):

Tinghan Jia ◽

Wu Qiao ◽

Qifeng Yao ◽

Wenhui Wu ◽

Ken Kaku

Keyword(s):

Topical Treatment ◽

Barrier Function ◽

Skin Diseases ◽

Morphological Differentiation ◽

Human Keratinocytes ◽

Human Epidermis ◽

Poly I:C ◽

Skin Barrier Function ◽

Mrna Levels ◽

Reconstructed Human Epidermis

Atopic dermatitis (AD) is a chronic inflammatory skin disease, which can cause skin barrier function damaged. Although co-incubation with docosahexaenoic acid (DHA) exerts a positive effect in deficient skin model, there is no study to investigate the effects of topical treatment with DHA in inflammatory reconstructed human epidermis (RHE) model. The effects of DHA on monolayer normal human epidermal keratinocyte (NHEK) cells were evaluated via CCK-8, qPCR and ELISA. The skin related barrier function was assessed by hematoxylin-eosin (HE) staining, western blot (WB), Immunohistofluorescence (IF) and ELISA in normal and inflammatory RHE models. DHA upregulated filaggrin and loricrin expression at mRNA levels in addition to suppress overexpression of TNF-α，IL-1α and IL-6 stimulated by poly I:C plus LPS (stimulation cocktail) in cultured NHEK cells. After topical treatment with DHA, cocktail induced inflammatory characteristics of skin diseases including barrier morphological, differentiation proteins and TSLP secretion, which were alleviated in RHE models. Supplementation with DHA can improved related barrier function and have anti-inflammation effects in monolayer keratinocytes and RHE models, which indicated that DHA may have a potential value for the treatment of inflammation-associate skin diseases.

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Treatment with Docosahexaenoic Acid Improves Epidermal Keratinocyte Differentiation and Ameliorates Inflammation in Human Keratinocytes and Reconstructed Human Epidermis Models

Molecules ◽

10.3390/molecules24173156 ◽

2019 ◽

Vol 24 (17) ◽

pp. 3156

Author(s):

Tinghan Jia ◽

Wu Qiao ◽

Qifeng Yao ◽

Wenhui Wu ◽

Ken Kaku

Keyword(s):

Docosahexaenoic Acid ◽

Topical Treatment ◽

Barrier Function ◽

Skin Diseases ◽

Human Epidermis ◽

Cell Counting ◽

Skin Barrier Function ◽

Enzyme Linked Immunosorbent Assay ◽

Epidermal Keratinocyte ◽

Reconstructed Human Epidermis

Atopic dermatitis (AD) is a chronic inflammatory skin disease that can cause skin barrier function damage. Although co-incubation with docosahexaenoic acid (DHA) exerts a positive effect on deficient skin models, no studies have investigated the effects of topical treatment with DHA in an inflammatory reconstructed human epidermis (RHE) model. The effects of DHA on monolayer normal human epidermal keratinocyte (NHEK) cells were evaluated using cell counting kit-8 (CCK-8), real-time quantitative polymerase chain reaction (qPCR), and enzyme-linked immunosorbent assay (ELISA). The skin-related barrier function was assessed using hematoxylin–eosin (HE) staining, Western blot (WB), immunohistofluorescence (IF), and ELISA in normal and inflammatory RHE models. Docosahexaenoic acid upregulated filaggrin and loricrin expression at mRNA levels in addition to suppressing overexpression of tumor necrosis factor-α (TNF-α), interleukin-α (IL-1α), and interleukin-6 (IL-6) stimulated by polyinosinic–polycytidylic acid (poly I:C) plus lipopolysaccharide (LPS) (stimulation cocktail) in cultured NHEK cells. After topical treatment with DHA, cocktail-induced inflammatory characteristics of skin diseases, including barrier morphology, differentiation proteins, and thymic stromal lymphopoietin (TSLP) secretion, were alleviated in RHE models. Supplementation with DHA can improve related barrier function and have anti-inflammation effects in monolayer keratinocytes and RHE models, which indicates that DHA may have potential value for the treatment of inflammation-associated skin diseases.

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Effect of retinaldehyde and calcium on epidermal barrier function in reconstructed human epidermis

Journal of the American Academy of Dermatology ◽

10.1016/j.jaad.2019.06.729 ◽

2019 ◽

Vol 81 (4) ◽

pp. AB197

Keyword(s):

Barrier Function ◽

Human Epidermis ◽

Epidermal Barrier ◽

Reconstructed Human Epidermis

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Stabilization of microtubules restores barrier function after cytokine-induced defects in reconstructed human epidermis

Journal of Dermatological Science ◽

10.1016/j.jdermsci.2018.04.008 ◽

2018 ◽

Vol 91 (1) ◽

pp. 87-96 ◽

Cited By ~ 5

Author(s):

Chiung-Yueh Hsu ◽

Nicolas Lecland ◽

Valérie Pendaries ◽

Cécile Viodé ◽

Daniel Redoulès ◽

...

Keyword(s):

Barrier Function ◽

Human Epidermis ◽

Reconstructed Human Epidermis ◽

Induced Defects

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Assessment of the Skin Barrier Function in the Reconstructed Human Epidermis using a Multimodal Approach at molecular, tissue and functional levels

The Analyst ◽

10.1039/d1an00465d ◽

2021 ◽

Author(s):

Joudi bakar ◽

Rime Michael-Jubeli ◽

Rindala El Khoury ◽

Sabrina Hamla ◽

Ali ASSI ◽

...

Keyword(s):

Barrier Function ◽

Functional Characterization ◽

Skin Barrier ◽

Human Epidermis ◽

Skin Barrier Function ◽

Multimodal Approach ◽

Reconstructed Human Epidermis

Reconstructed human epidermis models are used as epidermis alternatives in skin researches. It is necessary to provide molecular and functional characterization in order to assess these models. Our aim is...

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