Non-ionic amphiphilic biodegradable PEG-PLGA-PEG copolymer enhances gene delivery efficiency in rat skeletal muscle.
Naked plasmid DNA (pDNA)-based gene therapy has low delivery efficiency, and consequently, low therapeutic effect. We present a biodegradable nonionic triblock copolymer, PEG(13)-PLGA(10)-PEG(13), to enhance gene delivery efficiency in skeletal muscle. Effects of PEG(13)-PLGA(10)-PEG(13) on physicochemical properties of pDNA were evaluated by atomic force microscopy (AFM) imaging, gel electrophoresis and zeta-potential analysis. AFM imaging suggested a slightly compacted structure of pDNA when it was mixed with the polymer, while zeta-potential measurement indicated an increased surface potential of negatively charged pDNA. PEG(13)-PLGA(10)-PEG(13) showed a relatively lower toxicity compared to Pluronic P85 in a skeletal muscle cell line. The luciferase expression of pDNA delivered in 0.25% polymer solution was up to three orders of magnitude more than branched polyethylenimine (bPEI(25 k))/pDNA and three times more than that of naked pDNA five days after intramuscular administration. This in vivo gene delivery enhancement was also observed displaying a two-fold higher expression of human vascular endothelial growth factor (VEGF). Based on fluorescence labeled pDNA distribution, it is speculated that the greater diffusivity of PEG(13)-PLGA(10)-PEG(13)/pDNA compared to bPEI(25 k)/pDNA accounts for better transfection efficiency in vivo. To summarize, combining PEG(13)-PLGA(10)-PEG(13) with pDNA possesses the potential to improve gene delivery efficiency in skeletal muscle.
Chang
Chien-Wen
CW
Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT 84112-5820, USA.
Choi
Donghoon
D
Kim
Won Jong
WJ
Yockman
James W
JW
Christensen
Lane V
LV
Kim
Yong-Hee
YH
Kim
Sung Wan
SW
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004587:Electrophoresis, Agar Gel; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases; D008297:Male; D051379:Mice; D018625:Microscopy, Atomic Force; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism*; D011060:Poloxalene / Q000633:toxicity; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000633:toxicity; D011094:Polyethyleneimine / Q000737:chemistry; D011098:Polyglactin 910 / Q000737:chemistry* / Q000633:toxicity; D051381:Rats; D017207:Rats, Sprague-Dawley; D013499:Surface Properties; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000235:genetics
C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C422648:pluronic block copolymer p85; C520330:polyethylene glycol-poly(lactic-co-glycolic acid)-polyethylene glycol; D011098:Polyglactin 910; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; D011060:Poloxalene; D008156:Luciferases