title: Paracrine Activity from Adipose-Derived Stem Cells on In Vitro Wound Healing in Human Tympanic Membrane Keratinocytes.
abstract: Stem cell therapies for tympanic membrane repair have shown initial experimental success using mesenchymal stem cells in rat models to promote healing; however, the mechanisms providing this benefit are not known. We investigated in vitro the paracrine effects of human adipose-derived stem cells (ADSCs) on wound healing mechanisms for human tympanic membrane-derived keratinocytes (hTM) and immortalized human keratinocytes (HaCaT). ADSC conditioned media (CMADSC) were assessed for paracrine activity on keratinocyte proliferation and migration, with hypoxic conditions for ADSC culture used to generate contrasting effects on cytokine gene expression. Keratinocytes cultured in CMADSCshowed a significant increase in cell number compared to serum-free cultures and further significant increases in hypoxic CMADSC. Assessment of ADSC gene expression on a cytokine array showed a range of wound healing cytokines expressed and under stringent hypoxic and serum-free conditions was upregulated (VEGF A, MMP9, Tissue Factor, PAI-1) or downregulated (CXCL5, CCL7, TNF-α). Several of these may contribute to the activity of conditioned media on the keratinocytes with potential applications in TM perforation repair. VEGFA protein was confirmed by immunoassay to be increased in conditioned media. Together with gene regulation associated with hypoxia in ADSCs, this study has provided several strong leads for a stem cell-derived approach to TM wound healing.
authors.affiliation: 1 Ear Science Institute Australia , Nedlands, Australia .|2 School of Veterinary and Life Sciences, Murdoch University , Murdoch, Australia .
authors.forename: Huan Ting
authors.initials: HT
authors.lastname: Ong
authors.affiliation: 1 Ear Science Institute Australia , Nedlands, Australia .|3 Ear Sciences Centre, University of Western Australia , Perth, Australia .
authors.forename: Sharon L
authors.initials: SL
authors.lastname: Redmond
authors.affiliation: 1 Ear Science Institute Australia , Nedlands, Australia .|3 Ear Sciences Centre, University of Western Australia , Perth, Australia .
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Marano
authors.affiliation: 1 Ear Science Institute Australia , Nedlands, Australia .|3 Ear Sciences Centre, University of Western Australia , Perth, Australia .
authors.forename: Marcus D
authors.initials: MD
authors.lastname: Atlas
authors.affiliation: 4 Division of Surgery, Akershus University Hospital and University of Oslo , Oslo, Norway .|5 Centre for Clinical Research Västerås, University of Uppsala , Uppsala, Sweden .
authors.forename: Magnus
authors.initials: M
authors.lastname: von Unge
authors.affiliation: 4 Division of Surgery, Akershus University Hospital and University of Oslo , Oslo, Norway .
authors.forename: Peder
authors.initials: P
authors.lastname: Aabel
authors.affiliation: 1 Ear Science Institute Australia , Nedlands, Australia .|3 Ear Sciences Centre, University of Western Australia , Perth, Australia .|6 Centre for Cell Therapy and Regenerative Medicine, University of Western Australia , Perth, Australia .
authors.forename: Rodney J
authors.initials: RJ
authors.lastname: Dilley
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D015687:Cell Hypoxia / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism; D020869:Gene Expression Profiling; D063990:Gene Ontology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D015603:Keratinocytes / Q000187:drug effects / Q000473:pathology*; D019899:Paracrine Communication / Q000187:drug effects; D013234:Stem Cells / Q000166:cytology*; D014432:Tympanic Membrane / Q000473:pathology*; D014945:Wound Healing
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins
keywords: MSC; gene expression; growth factors; wound healing