title: Adipose-derived stem cells and keratinocytes in a chronic wound cell culture model: the role of hydroxyectoine.
abstract: Chronic wounds represent a major socio-economic problem in developed countries today. Wound healing is a complex biological process. It requires a well-orchestrated interaction of mediators, resident cells and infiltrating cells. In this context, mesenchymal stem cells and keratinocytes play a crucial role in tissue regeneration. In chronic wounds these processes are disturbed and cell viability is reduced. Hydroxyectoine (HyEc) is a membrane protecting osmolyte with protein and macromolecule stabilising properties. Adipose-derived stem cells (ASC) and keratinocytes were cultured with chronic wound fluid (CWF) and treated with HyEc. Proliferation was investigated using MTT test and migration was examined with transwell-migration assay and scratch assay. Gene expression changes of basic fibroblast growth factor (b-FGF), vascular endothelial growth factor (VEGF), matrix metalloproteinases-2 (MMP-2) and MMP-9 were analysed by quantitative real-time polymerase chain reaction (qRT-PCR). CWF significantly inhibited proliferation and migration of keratinocytes. Addition of HyEc did not affect these results. Proliferation capacity of ASC was not influenced by CWF whereas migration was significantly enhanced. HyEc significantly reduced ASC migration. Expression of b-FGF, VEGF, MMP-2 and MMP-9 in ASC, and b-FGF, VEGF and MMP-9 in keratinocytes was strongly induced by chronic wound fluid. HyEc enhanced CWF induced gene expression of VEGF in ASC and MMP-9 in keratinocytes. CWF negatively impaired keratinocyte function, which was not influenced by HyEc. ASC migration was stimulated by CWF, whereas HyEc significantly inhibited migration of ASC. CWF induced gene expression of VEGF in ASC and MMP-9 in keratinocytes was enhanced by HyEc, which might partly be explained by an RNA stabilising effect of HyEc.
authors.affiliation: Clinic for Plastic and Reconstructive Surgery, Handsurgery, Burn Care Center, University of Witten/Herdecke, Cologne-Merheim Medical Center, Köln, Germany.
authors.forename: Oliver C
authors.initials: OC
authors.lastname: Thamm
authors.affiliation: Clinic for Plastic and Reconstructive Surgery, Handsurgery, Burn Care Center, University of Witten/Herdecke, Cologne-Merheim Medical Center, Köln, Germany.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Theodorou
authors.affiliation: Institute for Research in Operative Medicine (IFOM), University of Witten/Herdecke, Köln, Germany.
authors.forename: Ewa
authors.initials: E
authors.lastname: Stuermer
authors.affiliation: Clinic for Plastic and Reconstructive Surgery, Handsurgery, Burn Care Center, University of Witten/Herdecke, Cologne-Merheim Medical Center, Köln, Germany.
authors.forename: Max J
authors.initials: MJ
authors.lastname: Zinser
authors.affiliation: Institute for Research in Operative Medicine (IFOM), University of Witten/Herdecke, Köln, Germany.
authors.forename: Edmund A
authors.initials: EA
authors.lastname: Neugebauer
authors.affiliation: Clinic for Plastic and Reconstructive Surgery, Handsurgery, Burn Care Center, University of Witten/Herdecke, Cologne-Merheim Medical Center, Köln, Germany.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Fuchs
authors.affiliation: Department of Trauma and Orthopedic Surgery, University of Witten/Herdecke, Cologne-Merheim Medical Center, Köln, Germany.
authors.forename: Paola
authors.initials: P
authors.lastname: Koenen
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000599:Amino Acids, Diamino / Q000627:therapeutic use*; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured / Q000187:drug effects; D002908:Chronic Disease / Q000628:therapy; D048091:Guided Tissue Regeneration / Q000379:methods; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000502:physiology*; D014945:Wound Healing / Q000187:drug effects*
chemical_list: D000599:Amino Acids, Diamino; C121557:hydroxyectoine
keywords: Adipose-derived stem cells; Chronic wound; Gene expression; Hydroxyectoine; Keratinocytes; Migration; Proliferation; Wound fluid; Wound healing