title: Nerve and blood vessel growth in response to grafted dermis and cultured keratinocytes.
abstract: The aim of this study was to study innervation and angiogenesis in response to grafts of dermis and cultured keratinocytes using immunohistochemical techniques. In a porcine model, fresh autologous de-epidermalized dermis and cultured autologous keratinocytes were combined using a two-stage technique, to produce keratodermal grafts. Wounds were encased within skin graft chambers that prevented the influence of the surrounding skin. As grafts contracted, a peripheral rim of granulation tissue became exposed, allowing us to compare the wound bed beneath grafts with that beneath the raw granulating surface. Grafts were studied for 6 weeks. Angiogenesis was studied using antisera to von Willebrand factor to detect endothelial cells. Nerve growth was studied using antisera to S-100, a Schwann cell marker, and to four axonal markers: protein gene product 9.5, C-flanking peptide of neuropeptide Y, calcitonin gene-related peptide, and vasoactive intestinal peptide. In kerato-dermal grafts (n = 28), organization of blood vessels and nerve growth occurred only beneath areas with epidermal cover as compared with the surrounding granulation tissue. Initially, the immunoreactivity to von Willebrand factor was high, but in areas with epidermal cover it assumed a more orderly pattern with fewer blood vessels. Innervation was first detected by S-100 immunoreactivity seen at 1 to 2 weeks, closely followed by that to protein gene product 9.5 and much later to calcitonin gene-related peptide. C-flanking peptide of neuropeptide Y and vasoactive intestinal peptide immunoreactivity were detected in the wound depth surrounding large blood vessels at 4 to 6 weeks. In control wounds that had been either grafted with de-epidermalized dermis alone (n = 10) or allowed to granulate (n = 10), persistently there was high immunoreactivity to von Willebrand factor but minimal immunoreactivity to the neural markers. In conclusion, kerato-dermal grafts become innervated, and beneath their surface there is also vascular organization to resemble normal skin. Keratinocytes themselves may influence angiogenesis and innervation, as both processes failed to occur beneath granulating areas.
authors.affiliation: RAFT Institute of Plastic Surgery, Mount Vernon Hospital, Northwood, Middlesex, United Kingdom.
authors.forename: T
authors.initials: T
authors.lastname: Kangesu
authors.forename: S
authors.initials: S
authors.lastname: Manek
authors.forename: G
authors.initials: G
authors.lastname: Terenghi
authors.forename: X H
authors.initials: XH
authors.lastname: Gu
authors.forename: H A
authors.initials: HA
authors.lastname: Navsaria
authors.forename: J M
authors.initials: JM
authors.lastname: Polak
authors.forename: C J
authors.initials: CJ
authors.lastname: Green
authors.forename: I M
authors.initials: IM
authors.lastname: Leigh
mesh_terms: D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000032:analysis; D002478:Cells, Cultured; D006097:Granulation Tissue / Q000737:chemistry; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000166:cytology / Q000637:transplantation*; D018919:Neovascularization, Physiologic; D009478:Neuropeptide Y / Q000032:analysis; D010446:Peptide Fragments / Q000032:analysis; D010525:Peripheral Nerves / Q000254:growth & development; D009418:S100 Proteins / Q000032:analysis; D012867:Skin / Q000098:blood supply* / Q000737:chemistry / Q000294:innervation*; D016038:Skin Transplantation; D013552:Swine; D013869:Thiolester Hydrolases / Q000032:analysis; D043222:Ubiquitin Thiolesterase; D014660:Vasoactive Intestinal Peptide / Q000032:analysis; D014841:von Willebrand Factor / Q000032:analysis
chemical_list: D009478:Neuropeptide Y; D010446:Peptide Fragments; D009418:S100 Proteins; C048612:neuropeptide Y C-terminal flanking peptide; D014841:von Willebrand Factor; D014660:Vasoactive Intestinal Peptide; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase; D015740:Calcitonin Gene-Related Peptide