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: Daniel J Tilkorn, E Michele Davies, Effie Keramidaris, Aaron M Dingle, Yi-Wen Gerrand, Caroline J Taylor, Xiao Lian Han, Jason A Palmer, Anthony J Penington, Christina A Mitchell, Wayne A Morrison, Gregory J Dusting, Geraldine M Mitchell
• Journal: Biomaterials (Biomaterials) Vol. 33 Issue 15 Pg. 3868-79 (May 2012) ISSN: 1878-5905 [Electronic] Netherlands
• PMID: 22369961 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012 Elsevier Ltd. All rights reserved.

Chemical References

• Desmin
• Nitroso Compounds
• Platelet Endothelial Cell Adhesion Molecule-1
• 2,2'-(hydroxynitrosohydrazono)bis-ethanamine
• Proto-Oncogene Proteins c-akt
• Extracellular Signal-Regulated MAP Kinases

Topics

• Animals
• Blood Vessels (drug effects, growth & development)
• Cell Count
• Cell Hypoxia (drug effects)
• Cell Line
• Cell Size (drug effects)
• Cell Survival (drug effects)
• Desmin (metabolism)
• Extracellular Signal-Regulated MAP Kinases (metabolism)
• MAP Kinase Signaling System (drug effects)
• Mice
• Mice, SCID
• Models, Biological
• Myoblasts (cytology, drug effects, enzymology)
• Nitroso Compounds (pharmacology)
• Platelet Endothelial Cell Adhesion Molecule-1 (metabolism)
• Prosthesis Implantation
• Proto-Oncogene Proteins c-akt (metabolism)
• Rats
• Tissue Engineering (methods)
• Tissue Scaffolds