[Establishment of animal model for electroporation-mediated gene therapy in distraction osteogenesis of rabbit mandible].
OBJECTIVE
To evaluate the feasibility of electroporation-mediated transfection of recombinant plasmid to mandibular distraction area of rabbit in vivo.


METHODS
New-Zeland rabbit were employed. The mandible was distracted 3 days after operation at a rate of 0.8 mm per day for 7 days. The rabbits were randomly divided into 3 groups as group A (recombinant plasmid pIRES-VEGF165-EGFP), group B (recombinant plasmid plRES-VEGF165-EGFP) and group C (normal saline). The rabbits were sacrified at 3 hours, 1, 3, 7 and 14 d after injection respectively. The tissue at the distraction area was taken out for frozen section. The gene expression was assessed by the detection of expression of green fluorescence protein (GFP) using fluorescence microscope. The liver and kidney function test (ALT, AST, BUN, Scr) and the histological examination of heart, liver and kidney were also performed.


RESULTS
GFP was seen in the distraction area in group A and group B 3 hours after injection, which increased at the 1st day, reached peak value at the 3rd day, decreased at the 7th day and was very lower at 14th day. The GFP expression was much stronger in group A than in group B. GFP was not expressed in group C. There was no statistical difference in the concentration of ALT, AST, BUN and Scr in serum of rabbits among the three groups.


CONCLUSIONS
Electroporation-mediated transfection of recombinant plasmid can be expressed in the distraction area of rabbits, and there was no toxicity to the liver and kidney of rabbits. Electroporation could obviously improve transfection efficiency in vivo. It indicates that electroporation-mediated transfection of recombinant plasmid to distraction area tissue of rabbits is feasible.
Wu
Guo-Ping
GP
Department of Plastic Surgery of the Affiliated Hospital of Luzhou Medical College, Luzhou 646000, China.
Li
Sheng-Hua
SH
Li
De-Ping
DP
Yang
Zhi-Hui
ZH
He
Xiao-Chuan
XC
Liao
Yi
Y
Guo
Li
L
D000818:Animals; D053672:Electrochemotherapy; D005260:Female; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000235:genetics; D008297:Male; D008334:Mandible / Q000601:surgery*; D023421:Models, Animal; D019857:Osteogenesis, Distraction; D011817:Rabbits; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics
C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins