Topical Mineralocorticoid Receptor Blockade Limits Glucocorticoid-Induced Epidermal Atrophy in Human Skin  Eve Maubec, Cédric Laouénan, Lydia Deschamps,

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Topical Mineralocorticoid Receptor Blockade Limits Glucocorticoid-Induced Epidermal Atrophy in Human Skin  Eve Maubec, Cédric Laouénan, Lydia Deschamps, Van Tuan Nguyen, Isabelle Scheer-Senyarich, Anne-Catherine Wackenheim-Jacobs, Maud Steff, Stéphanie Duhamel, Sarah Tubiana, Nesrine Brahimi, Stéphanie Leclerc-Mercier, Béatrice Crickx, Claudine Perret, Selim Aractingi, Brigitte Escoubet, Xavier Duval, Philippe Arnaud, Frederic Jaisser, France Mentré, Nicolette Farman  Journal of Investigative Dermatology  Volume 135, Issue 7, Pages 1781-1789 (July 2015) DOI: 10.1038/jid.2015.44 Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Variations in epidermal thickness and keratinocyte proliferation in cultured human skin explants. Explants were incubated with solvent (control), mineralocorticoid receptor (MR) antagonist, clobetasol, or clobetasol+MR antagonist. (a) Photographs of tissue (bar=0.1mm). Effects of canrenoate (10μM): (b) epidermal thickness (μm); (c) clobetasol (100nM) compared with clobetasol+canrenoate; values from the same subject are linked, with dashed line for median; (d) KI67 index in the canrenoate series. Effects of eplerenone (10μM): (e) epidermal thickness; (f) eplerenone compared with clobetasol+eplerenone; (g) KI67 index in the eplerenone series. Graphs are box-and-whisker plot with median (rule inside the box), 75th and 25th percentile (top and bottom of box), 10th and 90th percentiles (bars projecting up and down) of epidermal thickness (in μm), and outlier values falling outside. Comparisons of treatments: Wilcoxon paired test. der, dermis; ep, epidermis; SC, stratum corneum. Journal of Investigative Dermatology 2015 135, 1781-1789DOI: (10.1038/jid.2015.44) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Effect of epithelial sodium channel (ENaC) blockade in human skin explants. Skin explants from seven subjects were incubated with solvent, phenamil (an ENaC blocker), clobetasol, or clobetasol+phenamil. (a) Photographs of tissue sections (bar=0.1mm). (b) Epidermal thickness: box-and-whisker plots as described in Figure 1. (c) Effect of clobetasol+ phenamil on epidermal thickness compared with clobetasol alone; values from samples issued from the same subject are linked and the dashed line shows the median value for each treatment. (d) Immunofluorescence photographs of tissue sections labeled with KI67 antibody (bar=0.1mm). (e) KI67 index; box-and-whisker plots as described in Figure 1. Differences were evaluated by Wilcoxon paired test. der, dermis; ep, epidermis; SC, stratum corneum. Journal of Investigative Dermatology 2015 135, 1781-1789DOI: (10.1038/jid.2015.44) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 SPIREPI trial: epidermal thickness determined on histological sections of skin biopsy (day 29) in healthy volunteers. (a) Histology (hematoxylin–eosin staining) of skin treated with each treatment (bar=0.5mm). (b) Plots of epidermal thickness (in μm) measured by histology in all healthy volunteers (n=23) according to treatment. Box-and-whisker plots as described in Figure 1. (c and d) Clobetasol compared with placebo in c and with clobetasol+spironolactone in d. Values from the same subject are linked and the dashed line shows the median value for each treatment. (e) Hatched box-and-whisker plots of epidermal thickness in the 17 healthy volunteers who were clobetasol responders (post hoc analysis). Differences were evaluated using Wilcoxon paired test. der, dermis; ep, epidermis; SC, stratum corneum. Journal of Investigative Dermatology 2015 135, 1781-1789DOI: (10.1038/jid.2015.44) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 SPIREPI trial: ultrasound measurement of whole-skin thickness in healthy volunteers according to treatment. (a) Representative high-resolution ultrasound imaging of epidermis+dermis thickness. White bar (1mm) extends over the epidermis+dermis. (b) Change in ultrasound measurement (in μm) of whole-skin thickness for all volunteers (n=23). (c) The post hoc analysis for the 17 clobetasol responders. Values are provided as difference in whole-skin thickness between measurements performed on day 1 (D1) and day 29 (D29). Box-and-whisker plots are as described in Figure 1. Differences were evaluated using Wilcoxon paired test. der, dermis; Ep, epidermis, hyp, hypodermis. Journal of Investigative Dermatology 2015 135, 1781-1789DOI: (10.1038/jid.2015.44) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions