Hassan Ait El Madani,a,b

Emmanuelle Tancrède-Bohin,c

Armand Bensussan,b

Anne Colonna,a

Alain Dupuy,d

Martine Bagot,b and Ana-Maria Penaa a

L’Oréal Research and Innovation, Aulnay sous Bois, France b Université Paris 7, Denis Diderot, Centre de Recherche sur la Peau, INSERM UMR-S-976, Paris, France c L’Oréal Research and Innovation, Centre de Recherche Bioclinique, Hôpital St. Louis, Paris, France d Université Rennes 1, CHU de Rennes, Service de Dermatologie, Rennes, France

Abstract

Multiphoton microscopy has emerged in the past decade as a promising tool for noninvasive skin imaging. Our aim was to evaluate the potential of multiphoton microscopy to detect topical corticosteroids side effects within the epidermis and to provide new insights into their dynamics. Healthy volunteers were topically treated with clobetasol propionate on a small region of their forearms under overnight occlusion for three weeks. The treated region of each patient was investigated at D0, D7, D15, D22 (end of the treatment), and D60. Our study shows that multiphoton microscopy allows for the detection of corticoid-induced epidermis modifications: thinning of stratum corneum compactum and epidermis, decrease of keratinocytes size, and changes in their morphology from D7 to D22. We also show that multiphoton microscopy enables in vivo three-dimensional (3-D) quantitative assessment of melanin content. We observe that melanin density decreases during treatment and almost completely disappears at D22. Moreover, these alterations are reversible as they are no longer present at D60. Our study demonstrates that multiphoton microscopy is a convenient and powerful tool for noninvasive 3-D dynamical studies of skin integrity and pigmentation.

© 2012 Society of Photo-Optical Instrumentation Engineers (SPIE). [DOI: 10.1117/1.JBO.17.2.026009] Keywords: three-dimensional imaging; second harmonic generation; two-photon excited fluorescence; in vivo multiphoton microscopy; corticosteroid; skin; melanin. Paper 11407 received Jul. 27, 2011; revised manuscript received Nov. 4, 2011; accepted for publication Dec. 19, 2011; published online Feb. 28, 2012.

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skin multiphoton