Angiomyogenesis using liposome based vascular endothelial growth factor-165 transfection with skeletal myoblast for cardiac repair.
We aim to investigate the feasibility and efficacy of cholesterol (Chol)+DOTAP liposome (CD liposome) based human vascular endothelial growth factor-165 (hVEGF(165)) gene transfer into human skeletal myoblasts (hSkM) for cardiac repair. The feasibility and efficacy of CD liposome for gene transfer with hSkM was characterized using plasmid carrying enhanced green fluorescent protein (pEGFP). Based on the optimized transfection procedure, hSkM were transfected with CD lipoplexes carrying plasmid-hVEGF(165) (CD-phVEGF(165)). The genetically modified hSkM were transplanted into rat heart model of acute myocardial infarction. Flow cytometry revealed that about 7.99% hSkM could be transfected with pEGFP. Based on the optimized transfection condition, transfected hSkM expressed hVEGF(165) up to day-18 (1.7+/-0.1ng/ml) with peak at day-2 (13.1+/-0.52ng/ml) with >85% cell viability. Animal studies revealed that reduced apoptosis, improved angiogenesis with blood flow in group-3 animal's heart were achieved as compared to group-1 and 2. Ejection fraction was best recovered in group-3 animals. The study demonstrates that though gene transfection efficiency using CD liposome mediated hVEGF(165) gene transfer with hSkM was low; hVEGF(165) gene expression efficiency was sufficient to induce neovascularization, improve blood flow and injured heart function.
Ye
Lei
L
National University Medical Institutes, National University of Singapore, Singapore.
Haider
Husnain Kh
HKh
Tan
RuSan
R
Su
LiPing
L
Law
Peter K
PK
Zhang
Wei
W
Sim
Eugene K W
EK
D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D002784:Cholesterol / Q000737:chemistry; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D016027:Heart Transplantation; D008081:Liposomes / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D032448:Myoblasts, Skeletal / Q000166:cytology* / Q000378:metabolism*; D018919:Neovascularization, Physiologic; D010316:Particle Size; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D051381:Rats; D012039:Regional Blood Flow; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D000644:Quaternary Ammonium Compounds; D042461:Vascular Endothelial Growth Factor A; D004815:Epidermal Growth Factor; D002784:Cholesterol; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane