Simultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA–induced metabolic stress in reconstructed human skin

Abstract Solar ultraviolet longwave UVA1 exposure of human skin has short-term consequences at cellular and molecular level, leading at long-term to photoaging. Following exposure, reactive oxygen species (ROS) are generated, inducing oxidative stress that might impair cellular metabolic activity. H...

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Autores principales:	Thi Phuong Lien Ung, Seongbin Lim, Xavier Solinas, Pierre Mahou, Anatole Chessel, Claire Marionnet, Thomas Bornschlögl, Emmanuel Beaurepaire, Françoise Bernerd, Ana-Maria Pena, Chiara Stringari
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/eec8ea68d4c94ca09d655154a5ef399d
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spelling	oai:doaj.org-article:eec8ea68d4c94ca09d655154a5ef399d2021-11-14T12:20:56ZSimultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA–induced metabolic stress in reconstructed human skin10.1038/s41598-021-00126-82045-2322https://doaj.org/article/eec8ea68d4c94ca09d655154a5ef399d2021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-00126-8https://doaj.org/toc/2045-2322Abstract Solar ultraviolet longwave UVA1 exposure of human skin has short-term consequences at cellular and molecular level, leading at long-term to photoaging. Following exposure, reactive oxygen species (ROS) are generated, inducing oxidative stress that might impair cellular metabolic activity. However, the dynamic of UVA1 impact on cellular metabolism remains unknown because of lacking adequate live imaging techniques. Here we assess the UVA1-induced metabolic stress response in reconstructed human skin with multicolor two-photon fluorescence lifetime microscopy (FLIM). Simultaneous imaging of nicotinamide adenine dinucleotide (NAD(P)H) and flavin adenine dinucleotide (FAD) by wavelength mixing allows quantifying cellular metabolism in function of NAD(P)+/NAD(P)H and FAD/FADH2 redox ratios. After UVA1 exposure, we observe an increase of fraction of bound NAD(P)H and decrease of fraction of bound FAD indicating a metabolic switch from glycolysis to oxidative phosphorylation or oxidative stress possibly correlated to ROS generation. NAD(P)H and FAD biomarkers have unique temporal dynamic and sensitivity to skin cell types and UVA1 dose. While the FAD biomarker is UVA1 dose-dependent in keratinocytes, the NAD(P)H biomarker shows no dose dependence in keratinocytes, but is directly affected after exposure in fibroblasts, thus reflecting different skin cells sensitivities to oxidative stress. Finally, we show that a sunscreen including a UVA1 filter prevents UVA1 metabolic stress response from occurring.Thi Phuong Lien UngSeongbin LimXavier SolinasPierre MahouAnatole ChesselClaire MarionnetThomas BornschlöglEmmanuel BeaurepaireFrançoise BernerdAna-Maria PenaChiara StringariNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Thi Phuong Lien Ung Seongbin Lim Xavier Solinas Pierre Mahou Anatole Chessel Claire Marionnet Thomas Bornschlögl Emmanuel Beaurepaire Françoise Bernerd Ana-Maria Pena Chiara Stringari Simultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA–induced metabolic stress in reconstructed human skin
description	Abstract Solar ultraviolet longwave UVA1 exposure of human skin has short-term consequences at cellular and molecular level, leading at long-term to photoaging. Following exposure, reactive oxygen species (ROS) are generated, inducing oxidative stress that might impair cellular metabolic activity. However, the dynamic of UVA1 impact on cellular metabolism remains unknown because of lacking adequate live imaging techniques. Here we assess the UVA1-induced metabolic stress response in reconstructed human skin with multicolor two-photon fluorescence lifetime microscopy (FLIM). Simultaneous imaging of nicotinamide adenine dinucleotide (NAD(P)H) and flavin adenine dinucleotide (FAD) by wavelength mixing allows quantifying cellular metabolism in function of NAD(P)+/NAD(P)H and FAD/FADH2 redox ratios. After UVA1 exposure, we observe an increase of fraction of bound NAD(P)H and decrease of fraction of bound FAD indicating a metabolic switch from glycolysis to oxidative phosphorylation or oxidative stress possibly correlated to ROS generation. NAD(P)H and FAD biomarkers have unique temporal dynamic and sensitivity to skin cell types and UVA1 dose. While the FAD biomarker is UVA1 dose-dependent in keratinocytes, the NAD(P)H biomarker shows no dose dependence in keratinocytes, but is directly affected after exposure in fibroblasts, thus reflecting different skin cells sensitivities to oxidative stress. Finally, we show that a sunscreen including a UVA1 filter prevents UVA1 metabolic stress response from occurring.
format	article
author	Thi Phuong Lien Ung Seongbin Lim Xavier Solinas Pierre Mahou Anatole Chessel Claire Marionnet Thomas Bornschlögl Emmanuel Beaurepaire Françoise Bernerd Ana-Maria Pena Chiara Stringari
author_facet	Thi Phuong Lien Ung Seongbin Lim Xavier Solinas Pierre Mahou Anatole Chessel Claire Marionnet Thomas Bornschlögl Emmanuel Beaurepaire Françoise Bernerd Ana-Maria Pena Chiara Stringari
author_sort	Thi Phuong Lien Ung
title	Simultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA–induced metabolic stress in reconstructed human skin
title_short	Simultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA–induced metabolic stress in reconstructed human skin
title_full	Simultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA–induced metabolic stress in reconstructed human skin
title_fullStr	Simultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA–induced metabolic stress in reconstructed human skin
title_full_unstemmed	Simultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA–induced metabolic stress in reconstructed human skin
title_sort	simultaneous nad(p)h and fad fluorescence lifetime microscopy of long uva–induced metabolic stress in reconstructed human skin
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/eec8ea68d4c94ca09d655154a5ef399d
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Simultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA–induced metabolic stress in reconstructed human skin

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