Impact of ultraviolet radiation on dermal and epidermal DNA damage in a human pigmented bilayered skin substitute
Benjamin Goyer
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorUlysse Pereira
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorBrice Magne
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorDanielle Larouche
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorSélia Kearns-Turcotte
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorPatrick J. Rochette
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département d'ophtalmologie et d'oto-rhino-laryngologie – chirurgie cervico-faciale, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorLudovic Martin
Service de Dermatologie, CHU d'Angers, et Institut MitoVasc (UMR INSERM 1083, UMR CNRS 6015), Université d'Angers, Angers, France
Search for more papers by this authorCorresponding Author
Lucie Germain
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Correspondence
Prof Lucie Germain, Loex, Aile-R, CHU de Québec, Université Laval, 1401, 18e rue, Québec G1J 1Z4, QC, Canada.
Email: [email protected]
Search for more papers by this authorBenjamin Goyer
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorUlysse Pereira
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorBrice Magne
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorDanielle Larouche
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorSélia Kearns-Turcotte
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorPatrick J. Rochette
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département d'ophtalmologie et d'oto-rhino-laryngologie – chirurgie cervico-faciale, Faculté de médecine, Université Laval, Québec, QC, Canada
Search for more papers by this authorLudovic Martin
Service de Dermatologie, CHU d'Angers, et Institut MitoVasc (UMR INSERM 1083, UMR CNRS 6015), Université d'Angers, Angers, France
Search for more papers by this authorCorresponding Author
Lucie Germain
Centre de recherche du CHU de Québec, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Québec, QC, Canada
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada
Correspondence
Prof Lucie Germain, Loex, Aile-R, CHU de Québec, Université Laval, 1401, 18e rue, Québec G1J 1Z4, QC, Canada.
Email: [email protected]
Search for more papers by this authorAbstract
Our laboratory has developed a scaffold-free cell-based method of tissue engineering to produce bilayered tissue-engineered skin substitutes (TESs) from epidermal and dermal cells. However, TES pigmentation is absent or heterogeneous after grafting, due to a suboptimal number of melanocytes in culture. Our objectives were to produce TESs with a sufficient quantity of melanocytes from different pigmentation phototypes (light and dark) to achieve a homogeneous color and to evaluate whether the resulting pigmentation was photoprotective against ultraviolet radiation (UVR)-induced DNA damage in the dermis and the epidermis. TESs were cultured using different concentrations of melanocytes (100, 200, and 1,500 melanocytes/mm2), and pigmentation was evaluated in vitro and after grafting onto an athymic mouse excisional model. Dermal and epidermal DNA damage was next studied, exposing pigmented TESs to 13 and 32.5 J/cm2 UVR in vitro. We observed that melanocyte cell density increased with culture time until reaching a plateau corresponding to the cell distribution of native skin. Pigmentation of melanocyte-containing TESs was similar to donor skin, with visible melanin transfer from melanocytes to keratinocytes. The amount of melanin in TESs was inversely correlated to the UVR-induced formation of cyclobutane pyrimidine dimer in dermal fibroblasts and keratinocytes. Our results indicate that the pigmentation conferred by the addition of melanocytes in TESs protects against UVR-induced DNA damage. Therefore, autologous pigmented TESs could ensure photoprotection after grafting.
CONFLICT OF INTEREST
The authors declare no conflicts of interest.
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