title: Wound-healing factors secreted by epidermal keratinocytes and dermal fibroblasts in skin substitutes.
abstract: Full-skin substitutes, epidermal substitutes, and dermal substitutes are currently being used to heal deep burns and chronic ulcers. In this study, we investigated which wound-healing mediators are released from these constructs and whether keratinocyte-fibroblast interactions are involved. Autologous skin substitutes were constructed from human keratinocytes, fibroblasts, and acellular donor dermis. Full-thickness skin was used to represent an autograft. Secretion of wound-healing mediators was investigated by means of protein array, enzyme-linked immunosorbent assay, neutralizing antibodies, and conditioned culture supernatants. Full-skin substitutes and autografts produce high amounts of inflammatory/angiogenic mediators (IL-6, CCL2, CXCL1, CXCL8, and sST2). Epidermal and dermal substitutes produced less of these proteins. Epidermal-derived proinflammatory cytokines interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha) were found to mediate synergistically the secretion of these wound-healing mediators (with the exception of sST2) from fibroblasts in dermal substitutes. The secretion of proinflammatory cytokines (IL-1alpha, TNF-alpha), chemokine/mitogen (CCL5) and angiogenic factor (vascular endothelial growth factor) by epidermal substitutes and tissue remodeling factors (tissue inhibitor of metalloproteinase-2, hepatocyte growth factor) by dermal substitutes was not influenced by keratinocyte-fibroblast interactions. The full-skin substitute has a greater potential to stimulate wound healing than epidermal or dermal substitutes. Both epidermal-derived IL-1alpha and TNF-alpha are required to trigger the release of dermal-derived inflammatory/angiogenic mediators from skin substitutes.
authors.affiliation: Department of Dermatology, VU University Medical Centre, Amsterdam, The Netherlands.
authors.forename: Sander W
authors.initials: SW
authors.lastname: Spiekstra
authors.forename: Melanie
authors.initials: M
authors.lastname: Breetveld
authors.forename: Thomas
authors.initials: T
authors.lastname: Rustemeyer
authors.forename: Rik J
authors.initials: RJ
authors.lastname: Scheper
authors.forename: Susan
authors.initials: S
authors.lastname: Gibbs
mesh_terms: D018929:Cell Culture Techniques; D018946:Chemokine CCL5 / Q000378:metabolism; D054360:Chemokine CXCL1 / Q000378:metabolism; D018920:Coculture Techniques; D000078404:Epidermal Cells; D005347:Fibroblasts / Q000502:physiology*; D006801:Humans; D007375:Interleukin-1 / Q000502:physiology*; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism / Q000502:physiology*; D016038:Skin Transplantation / Q000502:physiology; D017768:Skin, Artificial; D014182:Transplantation, Autologous; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*; D014945:Wound Healing / Q000502:physiology*
chemical_list: D018946:Chemokine CCL5; D054360:Chemokine CXCL1; D007375:Interleukin-1; D015850:Interleukin-6; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha