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. 2024 Apr;30(4):e13708.
doi: 10.1111/srt.13708.

Noninvasive evaluation of the skin barrier in reconstructed human epidermis using speckle analysis: Correlation with Raman microspectroscopy

Léa Habib  1   2 , Léa Abi Nassif  3 , Marie Abboud  3 , Rime Michael-Jubeli  2 , Ali Tfayli  2 , Roger Lteif  1
Affiliations

Noninvasive evaluation of the skin barrier in reconstructed human epidermis using speckle analysis: Correlation with Raman microspectroscopy

Léa Habib et al. Skin Res Technol. 2024 Apr.

Abstract

Background: Reconstructed epidermis models, obtained from 3D keratinocytes culture, have gained significant prominence as prototypes for safety and efficacy testing in skin research. To effectively evaluate these models, it is essential to perform molecular and functional characterization. The skin's barrier function is one of the essential aspects of the epidermis that needs to be assessed. A noninvasive method is thus required for the evaluation of the skin barrier in these models. With this perspective, the aim of this feasibility study is to apply the speckle technique for the assessment of the skin barrier in the Reconstructed Human Epidermis (RHE).

Materials and methods: Speckle analysis as well as Raman microspectroscopy were performed on RHE samples at two maturation days, D17 and D20.

Results: Between D17 and D20, our study showed an increase in various Raman parameters, including stratum corneum percentage, lateral lipid packing, lipid-to-protein ratio, and protein secondary structure. Furthermore, the degree of light polarization and the speckle grain size also increased over this period.

Conclusion: The speckle technique proved to be effective for evaluating the skin barrier in Reconstructed Human Epidermis (RHE) models. Comparison with Raman validates this approach and provides comprehensive molecular and functional characterization of reconstructive skin models.

Keywords: barrier function; degree of polarization; epidermal models; grain size; lipids; skin research.

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Figures

FIGURE 1
FIGURE 1
The correlation between the kinetic slopes of caffeine percutaneous penetration and maturation days of the RHE. RHE, reconstructed human epidermis.
FIGURE 2
FIGURE 2
(A) Speckle experimental set‐up and (B) Speckle image (400 pixel × 400 pixel) recorded of the RHE at D20 of maturation. ϴ is the angle between backscattered and incident light; D is the distance between the camera and the RHE sample. RHE, reconstructed human epidermis.
FIGURE 3
FIGURE 3
Examples of speckles image recorded of the RHE at (A) D17 and (B) D20 of maturation. RHE, reconstructed human epidermis.
FIGURE 4
FIGURE 4
Variation of (A) the percentage of stratum corneum (SC) contribution, (B) the lateral packing of lipids, and (C) the lipids‐to‐proteins ratio within the SC of the RHE during different maturation days. RHE, reconstructed human epidermis; SC, stratum corneum.
FIGURE 5
FIGURE 5
Variation of (A) Amide I/α, (B) Amide I/β, and (C) Amide I/(α+β) ratios within the SC of the RHE during different maturation days. RHE, reconstructed human epidermis; SC, stratum corneum.
FIGURE 6
FIGURE 6
Variation of DOPL within the RHE during maturation days. Error bars are the standard deviations. DOPL, degree of liner polarization; RHE, reconstructed human epidermis.
FIGURE 7
FIGURE 7
Variation of horizontal speckle grain size within the RHE during maturation days. RHE, reconstructed human epidermis.
FIGURE 8
FIGURE 8
Variation of Raman and Speckle parameters between days 17 and 20 of RHE maturation. RHE, reconstructed human epidermis.
See this image and copyright information in PMC

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