High loading efficiency and sustained release of siRNA encapsulated in PLGA nanoparticles: Quality by design optimization and characterization |
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Authors: | Dongmei Cun,Ditte Krohn Jensen,Morten Jonas Maltesen,Matthew BunkerPaul Whiteside,David ScurrCamilla Foged,Hanne Mø rck Nielsen |
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Affiliation: | a Department of Pharmaceutics and Analytical Chemistry, University of Copenhagen, Copenhagen Ø, Denmark b Molecular Profiles Ltd, Nottingham, UK c Laboratory of Biophysics and Surface Analysis, School of Pharmacy, University of Nottingham, Nottingham, UK |
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Abstract: | Poly(dl-lactide-co-glycolide acid) (PLGA) is an attractive polymer for delivery of biopharmaceuticals owing to its biocompatibility, biodegradability and outstanding controlled release characteristics. The purpose of this study was to understand and define optimal parameters for preparation of small interfering RNA (siRNA)-loaded PLGA nanoparticles by the double emulsion solvent evaporation method and characterize their properties. The experiments were performed according to a 25−1 fractional factorial design based on five independent variables: The volume ratio between the inner water phase and the oil phase, the PLGA concentration, the sonication time, the siRNA load and the amount of acetylated bovine serum albumin (Ac-BSA) in the inner water phase added to stabilize the primary emulsion. The effects on the siRNA encapsulation efficiency and the particle size were investigated. The most important factors for obtaining an encapsulation efficiency as high as 70% were the PLGA concentration and the volume ratio whereas the size was mainly affected by the PLGA concentration. The viscosity of the oil phase was increased at high PLGA concentration, which explains the improved encapsulation by stabilization of the primary emulsion and reduction of siRNA leakage to the outer water phase. Addition of Ac-BSA increased the encapsulation efficiency at low PLGA concentrations. The PLGA matrix protected siRNA against nuclease degradation, provided a burst release of surface-localized siRNA followed by a triphasic sustained release for two months. These results enable careful understanding and definition of optimal process parameters for preparation of PLGA nanoparticles encapsulating high amounts of siRNA with immediate and long-term sustained release properties. |
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Keywords: | Ac-BSA, acetylated bovine serum albumin AFM, atomic force microscopy ANOVA, analysis of variance DCM, dichloromethane DEPC, diethyl pyrocarbonate DOE, design of experiments DOTAP, dioleyltrimethylammoniumpropane EGFP, enhanced green fluorescent protein FBS, fetal bovine serum FDA, Food and Drug Administration FFD, fractional factorial design FLuc, firefly luciferase LF, lipofectamine PBS, phosphate-buffered saline PCS, photon correlation spectroscopy PDI, polydispersity index PEI, polyethyleneimine PLGA, poly( smallcaps" >dl-lactide-co-glycolide acid) PVA, polyvinylalcohol siRNA, small interfering RNA ToF-SIMS, time-of-flight secondary ion mass spectrometry |
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