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Characterization of the structure of human skin substitutes by infrared microspectroscopy

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Abstract

The skin acts mainly as a protective barrier from the external environment, thanks to the stratum corneum which is the outermost layer of the skin. As in vitro tests on skin are essential to elaborate new drugs, the development of skin models closer to reality becomes essential. It is now possible to produce in vitro human skin substitutes through tissue engineering by using the self-assembly method developed by the Laboratoire d’Organogénèse Expérimentale. In the present work, infrared microspectroscopy imaging analyses were performed to get in-depth morpho-spectral characterization of the three characteristic layers of human skin substitutes and normal human skin, namely the stratum corneum, living epidermis, and dermis. An infrared spectral analysis of the skin is a powerful tool to gain information on the order and conformation of the lipid chains and the secondary structure of proteins. On one hand, the symmetric stretching mode of the lipid methylene groups (2,850 cm−1) is sensitive to the acyl chain conformational order. The evolution profile of the frequency of this vibrational mode throughout the epidermis suggests that lipids in the stratum corneum are more ordered than those in the living epidermis. On the other hand, the frequencies of the infrared components underneath the envelop of the amide I band provide information about the overall protein conformation. The analysis of this mode establishes that the proteins essentially adopt an α-helix conformation in the epidermis, probably associated with the presence of keratin, while modifications of the protein content are observed in the dermis (extracellular matrix made of collagen). Finally, the lipid organization, as well as the protein composition in the different layers, is similar for human skin substitutes and normal human skin, confirming that the substitutes reproduce essential features of real skin and are appropriate biomimetics.

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Acknowledgments

The authors acknowledge financial support from the National Science and Engineering Research Council (NSERC-Canada) and the Canadian Institutes for Health Research (CIHR) through their joint Collaborative Health Research Program and the Centre Québécois sur les Matériaux Fonctionnels. They also thank Jessica Jean for technical support during the tissue engineering experiments and Michel Pézolet for helpful discussions.

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Authors and Affiliations

  1. Laboratoire d’Ingénierie de Surface (LIS), Département de Génie des Mines, de la Métallurgie et des Matériaux, Centre de Recherche sur les Matériaux Avancés (CERMA), Université Laval, 1065 avenue de la médecine, Québec, QC, G1V 0A6, Canada

    Marie Leroy & Gaétan Laroche

  2. Centre de Recherche du Centre Hospitalier Universitaire de Québec (CRCHUQ), Hôpital St-François d’Assise, 10 rue de l’Espinay, Québec, QC, G1L 3L5, Canada

    Marie Leroy & Gaétan Laroche

  3. Centre LOEX de l’Université Laval, Génie Tissulaire et Régénération: LOEX-Centre de Recherche du Centre Hospitalier Universitaire de Québec (QC), Hôpital de l’Enfant Jesus, 1401, 18e rue, Québec, QC, G1J 1Z4, Canada

    Marie Leroy & Roxane Pouliot

  4. Département de Chimie, Regroupement québécois sur la fonction, la structure et l’ingénierie des protéines (PROTEO), CERMA, Université Laval, 1045 avenue de la médecine, Québec, QC, G1V 0A6, Canada

    Marie Leroy & Michèle Auger

  5. Center for Self-Assembled Chemical Structures (CSACS), Département de Chimie, Université de Montréal, Québec, QC, H3T 1J4, Canada

    Michel Lafleur

Authors
  1. Marie Leroy
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  2. Michel Lafleur
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  3. Michèle Auger
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  4. Gaétan Laroche
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  5. Roxane Pouliot
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Corresponding authors

Correspondence to Gaétan Laroche or Roxane Pouliot.

Additional information

Published in the topical collection Morpho-Spectral Imaging with guest editors Cyril Petibois and Yeukuang Hwu.

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Leroy, M., Lafleur, M., Auger, M. et al. Characterization of the structure of human skin substitutes by infrared microspectroscopy. Anal Bioanal Chem 405, 8709–8718 (2013). https://doi.org/10.1007/s00216-013-7103-y

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  • DOI: https://doi.org/10.1007/s00216-013-7103-y

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