Ex vivo non-viral vector-mediated neurotrophin-3 gene transfer to olfactory ensheathing glia: effects on axonal regeneration and functional recovery after implantation in rats with spinal cord injury.
OBJECTIVE
Combine olfactory ensheathing glia (OEG) implantation with ex vivo non-viral vector-based neurotrophin-3 (NT-3) gene therapy in attempting to enhance regeneration after thoracic spinal cord injury (SCI).


METHODS
Primary OEG were transfected with cationic liposome-mediated recombinant plasmid pcDNA3.1(+)-NT3 and subsequently implanted into adult Wistar rats directly after the thoracic spinal cord (T9) contusion by the New York University impactor. The animals in 3 different groups received 4x10(5) OEG transfected with pcDNA3.1(+)-NT3 or pcDNA3.1(+) plasmids, or the OEGs without any plasmid transfection, respectively; the fourth group was untreated group, in which no OEG was implanted.


RESULTS
NT-3 production was seen increased both ex vivo and in vivo in pcDNA3.1(+)-NT3 transfected OEGs. Three months after implantation of NT-3-transfected OEGs, behavioral analysis revealed that the hindlimb function of SCI rats was improved. All spinal cords were filled with regenerated neurofilament-positive axons. Retrograde tracing revealed enhanced regenerative axonal sprouting.


CONCLUSIONS
Non-viral vector-mediated genetic engineering of OEG was safe and more effective in producing NT-3 and promoting axonal outgrowth followed by enhancing SCI recovery in rats.
Wu
Jun
J
Department of Orthopedics, Traumatic Orthopedic Institute of PLA, Beijing Army General Hospital, Beijing 100700, China.
Sun
Tian-Sheng
TS
Ren
Ji-Xin
JX
Wang
Xian-Zhang
XZ
D000818:Animals; D000831:Animals, Newborn; D016380:Brain Tissue Transplantation / Q000379:methods*; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000627:therapeutic use; D004195:Disease Models, Animal; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D006085:Graft Survival / Q000235:genetics; D020439:Growth Cones / Q000378:metabolism / Q000648:ultrastructure; D009416:Nerve Regeneration / Q000235:genetics*; D009457:Neuroglia / Q000378:metabolism / Q000637:transplantation*; D020933:Neurotrophin 3 / Q000096:biosynthesis / Q000235:genetics*; D009830:Olfactory Bulb / Q000166:cytology / Q000637:transplantation; D010243:Paralysis / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy; D010957:Plasmids / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D020127:Recovery of Function / Q000235:genetics; D013119:Spinal Cord Injuries / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D016896:Treatment Outcome; D015854:Up-Regulation / Q000235:genetics
D004274:DNA, Recombinant; D020933:Neurotrophin 3