title: The in vitro preconditioning of myoblasts to enhance subsequent survival in an in vivo tissue engineering chamber model.
abstract: The effects of in vitro preconditioning protocols on the ultimate survival of myoblasts implanted in an in vivo tissue engineering chamber were examined. In vitro testing: L6 myoblasts were preconditioned by heat (42 °C; 1.5 h); hypoxia (<8% O(2); 1.5 h); or nitric oxide donors: S-nitroso-N-acetylpenicillamine (SNAP, 200 μM, 1.5 h) or 1-[N-(2-aminoethyl)-N-(2-aminoethyl)amino]-diazen-1-ium-1,2-diolate (DETA-NONOate, 500 μM, 7 h). Following a rest phase preconditioned cells were exposed to 24 h hypoxia, and demonstrated minimal overall cell loss, whilst controls (not preconditioned, but exposed to 24 h hypoxia) demonstrated a 44% cell loss. Phosphoimmunoblot analysis of pro-survival signaling pathways revealed significant activation of serine threonine kinase Akt with DETA-NONOate (p < 0.01) and heat preconditioning (p < 0.05). DETA-NONOate also activated ERK 1/2 signaling (p < 0.05). In vivo implantation: 100,000 preconditioned (heat, hypoxia, or DETA-NONOate) myoblasts were implanted in SCID mouse tissue engineering chambers. 100,000 (not preconditioned) myoblasts were implanted in control chambers. At 3 weeks, morphometric assessment of surviving myoblasts indicated myoblast percent volume (p = 0.012) and myoblasts/mm(2) (p = 0.0005) overall significantly increased in preconditioned myoblast chambers compared to control, with DETA-NONOate-preconditioned myoblasts demonstrating the greatest increase in survival (p = 0.007 and p = 0.001 respectively). DETA-NONOate therefore has potential therapeutic benefits to significantly improve survival of transplanted cells.
authors.affiliation: Department of Plastic Surgery, Burn Center, Hand Center, BG-University-Hospital Bergmannsheil, Ruhr-University Bochum, Germany.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Tilkorn
authors.forename: E Michele
authors.initials: EM
authors.lastname: Davies
authors.forename: Effie
authors.initials: E
authors.lastname: Keramidaris
authors.forename: Aaron M
authors.initials: AM
authors.lastname: Dingle
authors.forename: Yi-Wen
authors.initials: YW
authors.lastname: Gerrand
authors.forename: Caroline J
authors.initials: CJ
authors.lastname: Taylor
authors.forename: Xiao Lian
authors.initials: XL
authors.lastname: Han
authors.forename: Jason A
authors.initials: JA
authors.lastname: Palmer
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Penington
authors.forename: Christina A
authors.initials: CA
authors.lastname: Mitchell
authors.forename: Wayne A
authors.initials: WA
authors.lastname: Morrison
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Dusting
authors.forename: Geraldine M
authors.initials: GM
authors.lastname: Mitchell
mesh_terms: D000818:Animals; D001808:Blood Vessels / Q000187:drug effects / Q000254:growth & development; D002452:Cell Count; D015687:Cell Hypoxia / Q000187:drug effects; D002460:Cell Line; D048429:Cell Size / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003893:Desmin / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects; D051379:Mice; D016513:Mice, SCID; D008954:Models, Biological; D032446:Myoblasts / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D009603:Nitroso Compounds / Q000494:pharmacology; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D019919:Prosthesis Implantation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051381:Rats; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D003893:Desmin; D009603:Nitroso Compounds; D019408:Platelet Endothelial Cell Adhesion Molecule-1; C094210:2,2'-(hydroxynitrosohydrazono)bis-ethanamine; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases