Optimization of DNA delivery by three classes of hybrid nanoparticle/DNA complexes

Journal of Nanobiotechnology

Research

Sarala Pamujula¹  Tarun K Mandal¹  Dakshina Murthy Devanga Chinta¹  Ronald B Luftig²  Qiu Zhong²  Xin Yao²  Haifan Wang³ 

[1] College of Pharmacy, Xavier University of Louisiana, 70125, New Orleans, Louisiana, USA;Department of Microbiology Immunology and Parasitology, Louisiana State University Health Sciences Center, 70112, New Orleans, Louisiana, USA;Department of Microbiology Immunology and Parasitology, Louisiana State University Health Sciences Center, 70112, New Orleans, Louisiana, USA;Guangdong Food and Drug Vocational College, 510520, Guang Zhou, Guangdong, PR China;

关键词: Zeta Potential;    Cationic Lipid;    Protamine Sulphate;    Solvent Evaporation Method;    Viral Gene Therapy;   

DOI  :  10.1186/1477-3155-8-6

 received in 2009-11-24, accepted in 2010-02-24,  发布年份 2010

来源: Springer

【 摘 要 】

Plasmid DNA encoding a luciferase reporter gene was complexed with each of six different hybrid nanoparticles (NPs) synthesized from mixtures of poly (D, L-lactide-co-glycolide acid) (PLGA 50:50) and the cationic lipids DOTAP (1, 2-Dioleoyl-3-Trimethyammonium-Propane) or DC-Chol {3β-[N-(N', N'-Dimethylaminoethane)-carbamyl] Cholesterol}. Particles were 100-400 nm in diameter and the resulting complexes had DNA adsorbed on the surface (out), encapsulated (in), or DNA adsorbed and encapsulated (both). A luciferase reporter assay was used to quantify DNA expression in 293 cells for the uptake of six different NP/DNA complexes. Optimal DNA delivery occurred for 105 cells over a range of 500 ng - 10 μg of NPs containing 20-30 μg DNA per 1 mg of NPs. Uptake of DNA from NP/DNA complexes was found to be 500-600 times as efficient as unbound DNA. Regression analysis was performed and lines were drawn for DNA uptake over a four week interval. NP/DNA complexes with adsorbed NPs (out) showed a large initial uptake followed by a steep slope of DNA decline and large angle of declination; lines from uptake of adsorbed and encapsulated NPs (both) also exhibited a large initial uptake but was followed by a gradual slope of DNA decline and small angle of declination, indicating longer times of luciferase expression in 293 cells. NPs with encapsulated DNA only (in), gave an intermediate activity. The latter two effects were best seen with DOTAP-NPs while the former was best seen with DC-Chol-NPs. These results provide optimal conditions for using different hybrid NP/DNA complexes in vitro and in the future, will be tested in vivo.

【 授权许可】

CC BY   
© Zhong et al; licensee BioMed Central Ltd. 2010