Abstract
The development of dermal equivalents (DEs) for the treatment of burns has contributed toward efficient wound closure. A collagen-glycosaminoglycan DE (C-GAG) grafted with hair follicles converted a full-thickness wound to partial-thickness resulting in complete wound closure and restored hair. In this study we compared the ability of both intact pilosebaceous units (PSU) or truncated hair follicles (THF) to regenerate a multilayered epidermis in vitro when implanted into de-epidermalized dermis (DED) or C-GAG with the epidermis generated in vivo using C-CAG. Keratinocytes explanted from the outer root sheath of PSU and THF in both DED and C-GAG but only formed a multilayered epidermis with PSU in DED. PSU were more effective at forming multilayered epidermis in DED than THF. Both DED and C-GAG skin expressed proliferation (PCNA), differentiation (K1, K10), hyperproliferation (K6, K16), basal (K14), putative stem cell (p63), extracellular matrix protein (Collagen IV), mesenchymal (vimentin) and adherens junction (β-catenin) markers. These data suggest DEs supported initial maintenance of the implanted hair follicles, in particular PSU, and provide an excellent model with which to investigate the regulation of hair follicle progenitor epithelial cells during epidermal regeneration. Although neither PSU nor THF formed multilayered epidermis in C-CAG in vitro, hair follicles implanted into engrafted C-GAG on a burns patient resulted in epithelial regeneration and expression of proliferation and differentiation markers in a similar manner to that seen in vitro. However, the failure of C-GAG to support epidermal regeneration in vitro suggests in vivo factors are essential for full epidermal regeneration using C-GAG.
Highlights
Burn wounds excision followed by wound closure is the key to success for the treatment of large burn injuries and subsequent increase in survival rates [1]
We previously investigated the clinical outcome of early implantation of autologous pilosebaceous units (PSU) into a vascularised collagen-glycosaminoglycan DE (C-GAG) neo-dermis on a burns patient and examined the general histology of the grafted site 16 and 37 days post grafting [17]
We previously reported that implanting hair follicle micrografts into the same C-GAG neo-dermis on a full-thickness scalp burn wound gave rise to a normal multilayered, differentiated epidermis which was derived from outer root sheath (ORS) cells [17]
Summary
Burn wounds excision followed by wound closure is the key to success for the treatment of large burn injuries and subsequent increase in survival rates [1]. Split-thickness and allogenic skin grafts are typically used for the resurfacing of burn wounds. Skin models to study epithelial regeneration from the hair follicle permanent cover but, availability can be restricted due to sparse donor sites which can lead to pain and scar formation. The latter are temporary, subject to rejection, and pose a risk of viral transmission. Due to the flat dermo-epidermal junction, these sheets are fragile on full-thickness wounds and are prone to blistering influencing take rates [3]. Poor take rates can be improved by grafting these sheets in combination with a dermal component [4]
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