Gene delivery nanoparticles fabricated by supercritical fluid extraction of emulsions.
Non-viral polymeric gene delivery systems offer increased protection from nuclease degradation, enhanced plasmid DNA (pDNA) uptake, and controlled dosing to sustain the duration of pDNA action. Such gene delivery systems can be formulated from biocompatible and biodegradable polymers such as poly(D,L-lactic-co-glycolic) acid (PLGA). Experimental loading of hydrophilic macromolecules such as pDNA is low in polymeric particles. The study purpose was to develop a supercritical fluid extraction of emulsions (SFEE) process based on CO(2) for preparing pEGFP-PLGA nanoparticles with high plasmid loading and loading efficiency. Another objective was to determine the efficacy of pFlt23k, an anti-angiogenic pDNA capable of inhibiting vascular endothelial growth factor (VEGF) secretion, following nanoparticle formation using the SFEE process. Results indicated that the SFEE process allows high actual loading of pDNA (19.7%, w/w), high loading efficiency (>98%), and low residual solvents (<50 ppm), due to rapid particle formation from efficient solvent removal provided by the SFEE process. pFlt23K-PLGA nanoparticles were capable of in vitro transfection, significantly reducing secreted VEGF from human lung alveolar epithelial cells (A549) under normoxic and hypoxic conditions. pFlt23K-PLGA nanoparticles did not exhibit cytotoxicity and are of potential value in treating neovascular disorders wherein VEGF levels are elevated.
Mayo
Aaron S
AS
Department of Pharmaceutical Sciences, University of Nebraska Medical Center, Omaha, NE, United States.
Ambati
Balamurali K
BK
Kompella
Uday B
UB
D025924:Chromatography, Supercritical Fluid / Q000379:methods; D004247:DNA / Q000008:administration & dosage*; D004655:Emulsions; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000008:administration & dosage*; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry; D053758:Nanoparticles; D010957:Plasmids; D011100:Polyglycolic Acid / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011650:Pulmonary Alveoli / Q000166:cytology / Q000378:metabolism; D012997:Solvents / Q000737:chemistry; D014162:Transfection / Q000379:methods; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors / Q000378:metabolism
D004655:Emulsions; D012997:Solvents; D042461:Vascular Endothelial Growth Factor A; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D004247:DNA