In vivo multiphoton imaging for non‐invasive time course assessment of retinoids effects on human skin

Tancrède‐Bohin, Emmanuelle; Baldeweck, Thérèse; Brizion, Sébastien; Decencière, Étienne; Victorin, Steeve; Ngo, Blandine; Raynaud, Edouard; Souverain, Luc; Bagot, Martine; Pena, Ana‐Maria

doi:10.1111/srt.12877

Cited by 16 publications

(35 citation statements)

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“…In the study of retinoids effects under occlusion 51 , a clear decrease of melanin content in the living epidermis is evidenced on retinol-treated area (Fig. S8 ), whereas no significant modification is observed at the global level after one year of usual application conditions 52 (Fig. S9 a).…”

Section: Resultsmentioning

confidence: 97%

“…For example, a decrease in one layer could be counteracted by an increase in another layer, and hence, no change will be evidenced in the global density. It also allows evidencing slight modulations that are not detectable at the global level 52 .…”

Section: Resultsmentioning

confidence: 99%

“…The data for seasonal sun exposure (Fig. S9 ) are from retinoids 1 year-study and correspond to acquisitions performed at months M00 (March), M03 (June), M06 (September), M12 (March + 1 yr) on the control untreated forearm of volunteers; these have already been published 52 ).…”

Section: Resultsmentioning

confidence: 99%

“…Despite the small investigated epidermal volume, we have demonstrated that multiphoton global 3D epidermal melanin density is highly correlated with skin ITA value in constitutive pigmentation study, in agreement with ex vivo histology and HPLC chemical analysis results on skin samples with variable constitutive pigmentation 9 . Moreover, our results indicate that multiphoton imaging can provide quantitative data of interest in studying pigmentation modulations under different conditions (constitutive and acquired pigmentation, aging, natural UV exposure 52 or application of topical retinoids 51 , 52 or corticosteroids 45 ), but its ability to accurately assess melanin heterogeneity in skin pigmentary disorders remains to be studied.…”

Section: Discussionmentioning

confidence: 97%

“…Retinoids 1-year study (February 2011–April 2012, EC reference 2010/58; published 52 ) This study involved 30 female volunteers (50–65 y) with skin color ITA between 10° and 41° (ITA group III/IV). They applied Retinol 0.3% (RO, Retinol 0.3% cream, L’Oréal group) (n = 15) or all-trans-Retinoic acid 0.025% (RA, Retinoic acid 0.025% cream, Galderma) (n = 15) on one dorsal forearm versus a control product (white paraffin containing excipient, Bayer) on the other forearm for 1 year.…”

Section: Methodsmentioning

confidence: 99%

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In vivo melanin 3D quantification and z-epidermal distribution by multiphoton FLIM, phasor and Pseudo-FLIM analyses

Pena

Decencière

Brizion

et al. 2022

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Characterizing melanins in situ and determining their 3D z-epidermal distribution is paramount for understanding physiological/pathological processes of melanin neosynthesis, transfer, degradation or modulation with external UV exposure or cosmetic/pharmaceutical products. Multiphoton fluorescence intensity- and lifetime-based approaches have been shown to afford melanin detection, but how can one quantify melanin in vivo in 3D from multiphoton fluorescence lifetime (FLIM) data, especially since FLIM imaging requires long image acquisition times not compatible with 3D imaging in a clinical setup? We propose an approach combining (i) multiphoton FLIM, (ii) fast image acquisition times, and (iii) a melanin detection method called Pseudo-FLIM, based on slope analysis of autofluorescence intensity decays from temporally binned data. We compare Pseudo-FLIM to FLIM bi-exponential and phasor analyses of synthetic melanin, melanocytes/keratinocytes coculture and in vivo human skin. Using parameters of global 3D epidermal melanin density and z-epidermal distribution profile, we provide first insights into the in vivo knowledge of 3D melanin modulations with constitutive pigmentation versus ethnicity, with seasonality over 1 year and with topical application of retinoic acid or retinol on human skin. Applications of Pseudo-FLIM based melanin detection encompass physiological, pathological, or environmental factors-induced pigmentation modulations up to whitening, anti-photoaging, or photoprotection products evaluation.

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 99%

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In vivo melanin 3D quantification and z-epidermal distribution by multiphoton FLIM, phasor and Pseudo-FLIM analyses

Pena

Decencière

Brizion

et al. 2022

Sci Rep

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Research Techniques Made Simple: Emerging Imaging Technologies for Noninvasive Optical Biopsy of Human Skin

Lentsch

Baugh

Lee

et al. 2022

Journal of Investigative Dermatology

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In vivo imaging with a fast large-area multiphoton exoscope (FLAME) captures the melanin distribution heterogeneity in human skin

Vicente

Durkin

Shrestha

et al. 2022

Sci Rep

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Melanin plays a significant role in the regulation of epidermal homeostasis and photoprotection of human skin. The assessment of its epidermal distribution and overall content is of great interest due to its involvement in a wide range of physiological and pathological skin processes. Among several spectroscopic and optical imaging methods that have been reported for non-invasive quantification of melanin in human skin, the approach based on the detection of two-photon excited fluorescence lifetime distinguishes itself by enabling selective detection of melanin with sub-cellular resolution, thus facilitating its quantification while also resolving its depth-profile. A key limitation of prior studies on the melanin assessment based on this approach is their inability to account for the skin heterogeneity due to the reduced field of view of the images, which results in high dispersion of the measurement values. Pigmentation in both normal and pathological human skin is highly heterogeneous and its macroscopic quantification is critical for reliable measurements of the epidermal melanin distribution and for capturing melanin-related sensitive dynamic changes as a response to treatment. In this work, we employ a fast large-area multiphoton exoscope (FLAME), recently developed by our group for clinical skin imaging, that has the ability to evaluate the 3D distribution of epidermal melanin content in vivo macroscopically (millimeter scale) with microscopic resolution (sub-micron) and rapid acquisition rates (minutes). We demonstrate significant enhancement in the reliability of the melanin density and distribution measurements across Fitzpatrick skin types I to V by capturing the intra-subject pigmentation heterogeneity enabled by the large volumetric sampling. We also demonstrate the potential of this approach to provide consistent measurement results when imaging the same skin area at different times. These advances are critical for clinical and research applications related to monitoring pigment modulation as a response to therapies against pigmentary skin disorders, skin aging, as well as skin cancers.

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In vivo multiphoton imaging for non‐invasive time course assessment of retinoids effects on human skin

Abstract: This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

Cited by 16 publications

References 46 publications

In vivo melanin 3D quantification and z-epidermal distribution by multiphoton FLIM, phasor and Pseudo-FLIM analyses

In vivo melanin 3D quantification and z-epidermal distribution by multiphoton FLIM, phasor and Pseudo-FLIM analyses

Research Techniques Made Simple: Emerging Imaging Technologies for Noninvasive Optical Biopsy of Human Skin

In vivo imaging with a fast large-area multiphoton exoscope (FLAME) captures the melanin distribution heterogeneity in human skin

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