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A hydroxyethylated cholesterol-based cationic lipid for DNA delivery: effect of conditioning.
- Source :
-
International journal of pharmaceutics [Int J Pharm] 2004 Jun 18; Vol. 278 (1), pp. 143-63. - Publication Year :
- 2004
-
Abstract
- We have synthesised a novel cholesterol-based cationic lipid to promote DNA transfer in cells. This lipid, dimethyl hydroxyethyl aminopropane carbamoyl cholesterol iodide (DMHAPC-Chol) contains a biodegradable carbamoyl linker and a hydroxyethyl group in the polar amino head moiety and is characterised by NMR. Liposomes prepared from this lipid and dioleoyl phosphatidyl ethanolamine (DOPE) in equimolar proportion showed a weak cytotoxicity as revealed by MTT assays and are efficient to deliver plasmids DNA evaluated by the expression of reporter genes in vitro and in vivo. In this paper, we present an original method to determine the lipid concentration based on the colorimetric detection of the colipid DOPE and the measure of the molar ratio DOPE/cationic lipid in the liposome by FTIR spectroscopy. The liposomes and lipid/DNA complexes structures were characterized by transmission electron microscopy (TEM) and by quasi-elastic light scattering (QLS). TEM indicated that the complexes correspond to aggregates containing globular substructures with liposomes size. The method of immuno-gold labelling was used to detect plasmid in the complex and reveals the presence of DNA inside the aggregates. Transfection results showed efficient DNA transfer depending on the charge ratio and liposomes conditioning. Gel retardation results indicated that at a molar charge ratio between X = 1.5 and X = 2.5 (depending on the liposome conditioning), all DNA was taken by liposomes. We showed that conditioning by freeze-drying (lyophilization) facilitates storage and improves transfection efficiency. When the liposomes were lyophilized prior to DNA addition or when the complexes were subjected to freeze-thawing cycles, the obtained complexes showed a transfection with levels enhanced up to four and five-fold respectively for the lyophilized liposomes and freeze-thawed complexes. NMR was used to characterize the modifications under freezing which showed an effect on 31P spectra.<br /> (Copyright 2004 Elsevier B.V.)
- Subjects :
- Animals
Cations
Cell Line, Tumor
Cell Survival drug effects
Cell Survival genetics
Cholesterol genetics
Cholesterol toxicity
DNA genetics
DNA toxicity
Dose-Response Relationship, Drug
Female
Freeze Drying
Lipids administration & dosage
Lipids genetics
Lipids toxicity
Liposomes
Mice
Mice, Nude
Phosphatidylethanolamines administration & dosage
Phosphatidylethanolamines genetics
Phosphatidylethanolamines toxicity
Transfection methods
Cholesterol administration & dosage
DNA administration & dosage
Drug Delivery Systems methods
Subjects
Details
- Language :
- English
- ISSN :
- 0378-5173
- Volume :
- 278
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- International journal of pharmaceutics
- Publication Type :
- Academic Journal
- Accession number :
- 15158957
- Full Text :
- https://doi.org/10.1016/j.ijpharm.2004.03.003