Connexin mimetic peptides increase cell migration in scrape-wounded human organotypic skin models and alter gene expression of dermal fibroblasts

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Significant increases in skin wound healing rates occur by reducing connexin (Cx) mediated communication (CMC). We analysed effects of Gap27, a connexin mimetic peptide directly targeted to the extracellular loop of Cx43 and known to reduce CMC, on cell migration during wound closure of an in vitro organotypic human skin model, and expression of extra-cellular matrix (ECM) and adhesion genes during dermal fibroblast migration in eugylcaemic and hyperglycaemic conditions. Organotypic living skin equivalents derived from primary dermal fibroblasts and epidermal keratinocytes were created. Immunocytochemistry determined connexin expression profiles. Migration studies revealed rates of ‘wound closure’ over 78 hours in the presence or absence of 100 µM Gap27. Dermal fibroblasts were cultured in 5.5 mM or 25 mM glucose and treated for 24 hours with Gap27, or scrambled peptide (control) in triplicate. mRNA from migrating cells was subjected to PCR array analysis (SABiosciences), with gene expression fold changes = 2 considered regulatory.
Keratinocytes stratified in the model, expressing Cx43 in basal proliferating cells and limited Cx26 predominantly in upper layers reflecting profiles seen in vivo. Gap27 increased cell migration rates of keratinocytes and fibroblasts (P < 0.01). In euglycaemic migrating fibroblasts Gap27 upregulated thirty-four genes and downregulated 1 gene. Collagen 1a1, connective tissue growth factor, integrinß1 and matrix metalloproteinase 1 were significantly upregulated (P < 0.05). By contrast, under hyperglycaemia, Gap27 upregulated one gene and downregulated 9 genes, including collagens, integrins, fibronectin-1 and versican.
Connexin mimetic peptides blocked CMC and increased cell migration rates in this organotypic model. In euglycaemic conditions during fibroblast migration Gap27 induced upregulation of genes associated with ECM remodelling, but in high glucose these mechanisms appear impaired. Connexin mimetic peptides can improve migration and have therapeutic potential in wound healing.
Original languageEnglish
Title of host publicationJournal of Investigative Dermatology
PublisherElsevier B.V.
PagesS81
Number of pages1
Volume129
DOIs
Publication statusPublished - 9 Sep 2009

Keywords

  • connexin
  • wound-healing
  • diabetes
  • Gap 27

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