期刊
BIOMATERIALS
卷 33, 期 15, 页码 3868-3879出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2012.02.006
关键词
Cell preconditioning; In vitro test of cell survival; Implantation in an in vivo tissue engineering chamber; Morphometric assessment of cell survival
资金
- National Health and Medical Research Council (NHMRC)
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 degrees C; 1.5 h); hypoxia (<8% O-2; 1.5 h); or nitric oxide donors: S-nitroso-N-acetylpenicillamine (SNAP, 200 mu M, 1.5 h) or 1-[N-(2-aminoethyl)-N-(2-aminoethyl)aminol-diazen-l-ium-1,2-diolate (DETA-NONOate, 500 mu M, 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. (C) 2012 Elsevier Ltd. All rights reserved.
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