This study evaluated the structure and function of a dermal substitute composed of de-epidermalized, acellular human dermis, and tested its performance as part of a composite skin graft with cultured human keratinocytes. The acellular dermis retained much of the complex structure of native dermis, as demonstrated by immunostaining for collagen Types IV and VII. Collagen Types IV and VII were found on the papillary surface, and collagen Type IV was found at sites throughout the acellular dermis. To further demonstrate the functionality of the acellular dermis material, we seeded the papillary surface with cultured keratinocytes. In response to the architecture of the dermal papillae, the keratinocytes formed a three-dimensional epidermal structure that was several cell layers thick, and recreated the original skin rete ridges at the interface with the dermis. When these composite grafts were transplanted to athymic mice, host fibrovascular cells repopulated the acellular dermis. Some vessels grew into the acellular dermis along the original pathways of the human blood vessels, as demonstrated by co-localization of human and mouse collagen Type IV. The skin that developed from these grafts repigmented completely via passenger melanocytes from the keratinocyte cultures, was durable, and remained stable for more than 5 months.
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