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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2021-06-01
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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. |
format | Article |
id | doaj-art-34824dcff2fa4231a1bbe8693ddae864 |
institution | Kabale University |
issn | 1846-9558 |
language | English |
publishDate | 2021-06-01 |
publisher | Sciendo |
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series | Acta Pharmaceutica |
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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