Format

Send to

Choose Destination
J Tissue Eng Regen Med. 2019 Sep 10. doi: 10.1002/term.2959. [Epub ahead of print]

Impact of ultraviolet radiation on dermal and epidermal DNA damage in a human pigmented bilayered skin substitute.

Author information

1
Centre de recherche du CHU de Québec - Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX.
2
Département de chirurgie, Faculté de médecine, Université Laval, Québec, QC, Canada.
3
Service de Dermatologie, CHU d'Angers, et Institut MitoVasc (UMR INSERM 1083, UMR CNRS 6015), Université d'Angers, Angers, France.
4
Département d'ophtalmologie et d'oto-rhino-laryngologie - chirurgie cervico-faciale, Faculté de médecine, Université Laval, Québec, QC, Canada.

Abstract

Our laboratory has developed a scaffold-free cell-based method of tissue engineering to produce bilayered tissue-engineered skin substitutes (TES) 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, 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.

KEYWORDS:

Dewar photoproduct; cyclobutane pyrimidine dimer; pigmentation disorders; pyrimidine (6-4) pyrimidone photoproduct; skin color; tissue engineering; tissue-engineered medical products; ultraviolet rays

PMID:
31502756
DOI:
10.1002/term.2959

Supplemental Content

Full text links

Icon for Wiley
Loading ...
Support Center