Ultraviolet B irradiation-induced keratinocyte senescence and impaired development of 3D epidermal reconstruct

Ultraviolet B (UVB) induces morphological and functional changes of the skin. This study investigated the effect of UVB on keratinocyte senescence and the development of reconstructed human epidermis (RHE). Primary normal human keratinocytes (NHK) from juvenile foreskin were irradiated with UVB (30...

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Main Authors: Wattanapitayakul Suvara K., Chularojmontri Linda, Schäfer-Korting Monika
Format: Article
Language:English
Published: Sciendo 2021-06-01
Series:Acta Pharmaceutica
Subjects:
Online Access:https://doi.org/10.2478/acph-2021-0011
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author Wattanapitayakul Suvara K.
Chularojmontri Linda
Schäfer-Korting Monika
author_facet Wattanapitayakul Suvara K.
Chularojmontri Linda
Schäfer-Korting Monika
author_sort Wattanapitayakul Suvara K.
collection DOAJ
description Ultraviolet B (UVB) induces morphological and functional changes of the skin. This study investigated the effect of UVB on keratinocyte senescence and the development of reconstructed human epidermis (RHE). Primary normal human keratinocytes (NHK) from juvenile foreskin were irradiated with UVB (30 mJ cm−2) and these effects were compared to NHK that underwent senescence in the late passage. UVB enhanced the accumulation of reactive oxygen species (ROS) and halted cell replication as detected by BrdU cell proliferation assay. The senescence phenotype was evaluated by beta-galactosidase (β-gal) staining and qPCR of genes related to senescent regulation, i.e. p16INK4a, cyclin D2, and IFI27. Senescence induced by high dose UVB resulted in morphological changes, enhanced β-gal activity, elevated cellular ROS levels and reduced DNA synthesis. qPCR revealed differential expression of the genes regulated senescence. p16INK4a expression was significantly increased in NHK exposed to UVB whereas enhanced IFI27 expression was observed only in cultural senescence. The levels of cyclin D2 expression were not significantly altered either by UVB or long culturing conditions. UVB significantly induced the aging phenotype in keratinocytes and impaired epidermal development. RHE generated from UVB-irradiated keratinocytes showed a thinner cross-sectional structure and the majority of keratinocytes in the lower epidermis were degenerated. The 3D epidermis model is useful in studying the skin aging process.
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spelling doaj-art-34824dcff2fa4231a1bbe8693ddae8642025-02-02T00:31:54ZengSciendoActa Pharmaceutica1846-95582021-06-0171229330310.2478/acph-2021-0011acph-2021-0011Ultraviolet B irradiation-induced keratinocyte senescence and impaired development of 3D epidermal reconstructWattanapitayakul Suvara K.0Chularojmontri Linda1Schäfer-Korting Monika2Department of Pharmacology Faculty of Medicine, Srinakharinwirot University, Bangkok10110, ThailandDepartment of Preclinical Sciences Faculty of Medicine, Thammasat University, Klongluang, Pathumthani 12120, ThailandInstitute for Pharmacy, Freie Universität Berlin, 14195BerlinGermanyUltraviolet B (UVB) induces morphological and functional changes of the skin. This study investigated the effect of UVB on keratinocyte senescence and the development of reconstructed human epidermis (RHE). Primary normal human keratinocytes (NHK) from juvenile foreskin were irradiated with UVB (30 mJ cm−2) and these effects were compared to NHK that underwent senescence in the late passage. UVB enhanced the accumulation of reactive oxygen species (ROS) and halted cell replication as detected by BrdU cell proliferation assay. The senescence phenotype was evaluated by beta-galactosidase (β-gal) staining and qPCR of genes related to senescent regulation, i.e. p16INK4a, cyclin D2, and IFI27. Senescence induced by high dose UVB resulted in morphological changes, enhanced β-gal activity, elevated cellular ROS levels and reduced DNA synthesis. qPCR revealed differential expression of the genes regulated senescence. p16INK4a expression was significantly increased in NHK exposed to UVB whereas enhanced IFI27 expression was observed only in cultural senescence. The levels of cyclin D2 expression were not significantly altered either by UVB or long culturing conditions. UVB significantly induced the aging phenotype in keratinocytes and impaired epidermal development. RHE generated from UVB-irradiated keratinocytes showed a thinner cross-sectional structure and the majority of keratinocytes in the lower epidermis were degenerated. The 3D epidermis model is useful in studying the skin aging process.https://doi.org/10.2478/acph-2021-0011epidermiskeratinocytesultraviolet radiationskin agingsenescencereactive oxygen species
spellingShingle Wattanapitayakul Suvara K.
Chularojmontri Linda
Schäfer-Korting Monika
Ultraviolet B irradiation-induced keratinocyte senescence and impaired development of 3D epidermal reconstruct
Acta Pharmaceutica
epidermis
keratinocytes
ultraviolet radiation
skin aging
senescence
reactive oxygen species
title Ultraviolet B irradiation-induced keratinocyte senescence and impaired development of 3D epidermal reconstruct
title_full Ultraviolet B irradiation-induced keratinocyte senescence and impaired development of 3D epidermal reconstruct
title_fullStr Ultraviolet B irradiation-induced keratinocyte senescence and impaired development of 3D epidermal reconstruct
title_full_unstemmed Ultraviolet B irradiation-induced keratinocyte senescence and impaired development of 3D epidermal reconstruct
title_short Ultraviolet B irradiation-induced keratinocyte senescence and impaired development of 3D epidermal reconstruct
title_sort ultraviolet b irradiation induced keratinocyte senescence and impaired development of 3d epidermal reconstruct
topic epidermis
keratinocytes
ultraviolet radiation
skin aging
senescence
reactive oxygen species
url https://doi.org/10.2478/acph-2021-0011
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AT chularojmontrilinda ultravioletbirradiationinducedkeratinocytesenescenceandimpaireddevelopmentof3depidermalreconstruct
AT schaferkortingmonika ultravioletbirradiationinducedkeratinocytesenescenceandimpaireddevelopmentof3depidermalreconstruct