Transplantation of nanoparticle transfected skeletal myoblasts overexpressing vascular endothelial growth factor-165 for cardiac repair.
BACKGROUND
We investigated the feasibility and efficacy of polyethylenimine (PEI) based human vascular endothelial growth factor-165 (hVEGF165) gene transfer into human skeletal myoblasts (HSM) for cell based delivery to the infarcted myocardium.


RESULTS
Based on optimized transfection procedure using enhanced green fluorescent protein (pEGFP), HSM were transfected with plasmid-hVEGF165 (phVEGF165) carried by PEI (PEI-phVEGF165) nanoparticles. The transfected HSM were characterized for transfection and expression of hVEGF165 in vitro and transplanted into rat heart model of acute myocardial infarction (AMI): group-1=DMEM injection, group-2= HSM transplantation, group-3= PEI-phVEGF165-transfected HSM (PEI-phVEGF165 myoblast) transplantation. A total of 48 rats received cyclosporine injection from 3 days before and until 4 weeks after cell transplantation. Echocardiography was performed to assess the heart function. Animals were sacrificed for molecular and histological studies on the heart tissue at 4 weeks after treatment. Based on optimized transfection conditions, transfected HSM expressed hVEGF165 for 18 days with >90% cell viability in vitro. Apoptotic index was reduced in group-2 and group-3 as compared with group-1. Blood vessel density (x400) by immunostaining for PECAM-1 in group-3 was significantly higher (P=0.043 for both) as compared with group-1 and group-2 at 4 weeks. Regional blood flow (ml/min/g) in the left ventricular anterior wall was higher in group-3 (P=0.043 for both) as compared with group-1 and group-2. Improved ejection fraction was achieved in group-3 (58.44+/-4.92%) as compared with group-1 (P=0.004).


CONCLUSIONS
PEI nanoparticle mediated hVEGF165 gene transfer into HSM is feasible and safe. It may serve as a novel and efficient alternative for angiomyogenesis in cardiac repair.
Ye
Lei
L
National University Medical Institutes, National University of Singapore, Singapore.
Haider
Husnain Kh
HKh
Tan
RuSan
R
Toh
WeeChi
W
Law
Peter K
PK
Tan
WeeBeng
W
Su
LiPing
L
Zhang
Wei
W
Ge
RuoWen
R
Zhang
Yong
Y
Lim
YeanTeng
Y
Sim
Eugene K W
EK
D000818:Animals; D002470:Cell Survival / Q000235:genetics; D017690:Cell Transplantation / Q000379:methods; D005260:Female; D018014:Gene Transfer Techniques; D006801:Humans; D032448:Myoblasts, Skeletal / Q000502:physiology / Q000637:transplantation*; D009203:Myocardial Infarction / Q000235:genetics* / Q000601:surgery; D053758:Nanoparticles / Q000008:administration & dosage*; D010316:Particle Size; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A / Q000008:administration & dosage* / Q000096:biosynthesis / Q000235:genetics*
C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A