標題: Uptake and transfection efficiency of PEGylated cationic liposome-DNA complexes with and without RGD-tagging
作者: Majzoub, Ramsey N.
Chan, Chia-Ling
Ewert, Kai K.
Silva, Bruno F. B.
Liang, Keng S.
Jacovetty, Erica L.
Carragher, Bridget
Potter, Clinton S.
Safinya, Cyrus R.
電子物理學系
Department of Electrophysics
關鍵字: Gene therapy;Live cell imaging;Liposome;Nanopartide;Polyethylene glycol;RGD peptide
公開日期: 1-六月-2014
摘要: Steric stabilization of cationic liposome DNA (CL DNA) complexes is required for in vivo applications such as gene therapy. PEGylation (PEG: poly(ethylene glycol)) of CL DNA complexes by addition of PEG2000-lipids yields sterically stabilized nanoparticles but strongly reduces their gene delivery efficacy. PEGylation-induced weakening of the electrostatic binding of CL DNA nanoparticles to cells (leading to reduced uptake) has been considered as a possible cause, but experimental results have been ambiguous. Using quantitative live-cell imaging in vitro, we have investigated cell attachment and uptake of PEGylated CL DNA nanopartides with and without a custom synthesized RGD-peptide grafted to the distal ends of PEG2000-lipids. The RGD-tagged nanoparticles exhibit strongly increased cellular attachment as well as uptake compared to nanoparticles without grafted peptide. Transfection efficiency of RGD-tagged PEGylated CL DNA NPs increases by about an order of magnitude between NPs with low and high membrane charge density (sigma(M); the average charge per unit area of the membrane; controlled by the molar ratio of cationic to neutral lipid), even though imaging data show that uptake of RGD-tagged particles is only slightly enhanced by high am. This suggests that endosomal escape and, as a result, transfection efficiency of RGD-tagged NPs is facilitated by high sigma(M). We present a model describing the interactions between PEGylated CL DNA nanoparticles and the anionic cell membrane which shows how the PEG grafting density and membrane charge density affect adhesion of nanoparticles to the cell surface. (C) 2014 Elsevier Ltd. All rights reserved.
URI: http://dx.doi.org/10.1016/j.biomaterials.2014.03.007
http://hdl.handle.net/11536/24184
ISSN: 0142-9612
DOI: 10.1016/j.biomaterials.2014.03.007
期刊: BIOMATERIALS
Volume: 35
Issue: 18
起始頁: 4996
結束頁: 5005
顯示於類別:期刊論文


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