Modified nanoprecipitation method to fabricate DNA-loaded PLGA nanoparticles.
OBJECTIVE
The objective of this study was to formulate DNA-loaded poly(d,l-lactide-co-glycotide) (PLGA) nanoparticles by a modified nanoprecipitation method.


METHODS
DNA-loaded PLGA nanoparticles were prepared by the modified nanoprecipitation method and the double emulsion/solvent evaporation method. The characterizations of DNA-loaded nanoparticles such as entrapment efficiency, morphology, particle size, zeta potential, structural integrity of the loaded DNA, and stability of the loaded DNA in PLGA nanoparticles against DNase I, in vitro release, cell viability and in vitro transfection capability were investigated.


RESULTS
The resulted PLGA nanoparticles by the modified nanoprecipitation method had uniform spherical shape, narrow size distribution with average particles size near 200 nm, negative zeta potential of -12.6 mV at pH 7.4, and a sustained-release property in vitro. Plasmid DNA could be efficiently encapsulated into PLGA nanoparticles (> 95%) without affecting its intact conformation using this modified nanoprecipitation method, which was superior to the double emulsion/solvent evaporation method. The PLGA nanoparticles were much safer to A549 cell compared to commercial Lipofectamine 2000 and could successfully transfer plasmid-enhanced green fluorescent protein into A549 cells.


CONCLUSIONS
In conclusion, the modified nanoprecipitation method could be applied as an efficient way to fabricate DNA-loaded PLGA nanoparticles instead of the conventional double emulsion/solvent evaporation method.
Niu
Xiuming
X
Department of Pharmacy, Shandong Medical College, Ji'nan, Shandong Province, PR China.
Zou
Weiwei
W
Liu
Chunxi
C
Zhang
Na
N
Fu
Chunhua
C
D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D011232:Chemical Precipitation; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D003850:Deoxyribonuclease I / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry* / Q000633:toxicity; D004339:Drug Compounding / Q000379:methods*; D004355:Drug Stability; D004587:Electrophoresis, Agar Gel; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry* / Q000633:toxicity; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D053758:Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D010316:Particle Size; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D011100:Polyglycolic Acid / Q000737:chemistry* / Q000633:toxicity; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D012995:Solubility; D013499:Surface Properties; D014162:Transfection
D004337:Drug Carriers; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D004247:DNA; D003850:Deoxyribonuclease I