Abstract
Currently, engineered skin substitutes (ESS) are unable to regenerate cutaneous appendages. Recent studies have shown that skin-derived precursors (SKPs), which are extensively available, have the potential to induce hair follicle neogenesis. Here, we demonstrate that ESS consisting of culture-expanded SKPs and epidermal stem cells (Epi-SCs) reconstitute the skin with hair follicle regeneration after grafting into nude mice. SKPs seeded in a C-GAG matrix proliferated and expressed higher levels of hair induction signature genes—such as Akp2, Sox2, CD133 and Bmp6—compared to dermal fibroblasts. Moreover, when ESS prepared by seeding a mixture of culture-expanded murine SKPs and human adult Epi-SCs into a C-GAG matrix was grafted into full-thickness skin wounds in nude mice, black hairs were generated within 3 weeks. Immunofluorescence analysis showed that the SKPs were localized to the dermal papillae of the newly-formed hair follicle. Our results indicate that SKPs can serve as the hair-inductive cells in ESS to furnish it with hair genesis potential
Highlights
The immunofluorescence analysis of the wound tissue indicated that the regenerated epidermis, hair follicles and sebaceous glands were largely derived from the implanted epidermal stem cells (Epi-SCs) (Figure 6D), while the donor-derived skin-derived precursors (SKPs) contributed to the dermal papilla (DP) in the neogenic hair follicles and numerous cells in the newly formed dermis
We further examined the structure of the hair shafts of the follicles regenerated from grafts of Engineered skin substitutes (ESS) consisting of SKPs and neonatal mouse epidermal cells in C-GAG under a microscope, and compared them with hairs collected from the back of adult wild-type C57 mice
ESS capable of fully restoring the structure and function the injured skin To hasdevelop been the in tissue engineering
Summary
Recent studies indicate that multipotent skin-derived precursors (SKPs)—present in the dermis of neonatal or adult human and mouse skin—have been shown to induce hair follicle neogenesis when transplanted, indicating their potential applications in hair follicle regeneration and ESS [14,15]. The survival and hair follicle-inductive ability of SKPs in C-GAG have not been studied. ESSs populated with culture-expanded human adult epidermal stem cells (Epi-SCs) and culture-expanded SKPs, but not dermal fibroblasts, regenerated the hair follicle and sebaceous gland after transplantation into wounds in nude mice. Our results indicate that SKPs can serve as hair-inductive cells in ESS in order to enhance its ability in appendage regeneration
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