Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells

Michelle Fraga1,2, Fernanda Bruxel1, Valeska Lizzi Lagranha2,3, Helder Ferreira Teixeira1, Ursula Matte2,31Post Graduation Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, 2Gene Therapy Center, Experimental Research Center, Hospital de Cl&iacute;nicas de Porto A...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Fraga M, Bruxel F, Lagranha VL, Teixeira HF, Matte U
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2011
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/36b64bf081f9498484b02dcd3bd1ec0d
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:36b64bf081f9498484b02dcd3bd1ec0d
record_format	dspace
spelling	oai:doaj.org-article:36b64bf081f9498484b02dcd3bd1ec0d2021-12-02T05:32:49ZInfluence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells1176-91141178-2013https://doaj.org/article/36b64bf081f9498484b02dcd3bd1ec0d2011-10-01T00:00:00Zhttp://www.dovepress.com/influence-of-phospholipid-composition-on-cationic-emulsionsdna-complex-a8432https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Michelle Fraga1,2, Fernanda Bruxel1, Valeska Lizzi Lagranha2,3, Helder Ferreira Teixeira1, Ursula Matte2,31Post Graduation Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, 2Gene Therapy Center, Experimental Research Center, Hospital de Cl&iacute;nicas de Porto Alegre, 3Post Graduation Program in Genetics and Molecular Biology, Universidade Federal do Rio Grande do Sul, Porto Alegre, BrazilBackground: Cationic nanoemulsions have been recently considered as potential delivery systems for nucleic acids. This study reports the influence of phospholipids on the properties of cationic nanoemulsions/DNA plasmid complexes.Methods: Nanoemulsions composed of medium-chain triglycerides, stearylamine, egg lecithin or isolated phospholipids, ie, DSPC, DOPC, DSPE, or DOPE, glycerol, and water were prepared by spontaneous emulsification. Gene transfer to Hep G2 cells was analyzed using real-time polymerase chain reaction.Results: The procedure resulted in monodispersed nanoemulsions with a droplet size and zeta potential of approximately 250 nm and +50 mV, respectively. The complexation of cationic nanoemulsions with DNA plasmid, analyzed by agarose gel retardation assay, was complete when the complex was obtained at a charge ratio of &ge;1.0. In these conditions, the complexes were protected from enzymatic degradation by DNase I. The cytotoxicity of the complexes in Hep G2 cells, evaluated by MTT assay, showed that an increasing number of complexes led to progressive toxicity. Higher amounts of reporter DNA were detected for the formulation obtained with the DSPC phospholipid. Complexes containing DSPC and DSPE phospholipids, which have high phase transition temperatures, were less toxic in comparison with the formulations obtained with lecithin, DOPC, and DOPE.Conclusion: The results show the effect of the DNA/nanoemulsion complexes composition on the toxicity and transfection results.Keywords: plasmids, cationic nanoemulsions, phospholipids, physicochemical characterization, gene transfer, stearylamineFraga MBruxel FLagranha VLTeixeira HFMatte UDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2011, Iss default, Pp 2213-2220 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Fraga M Bruxel F Lagranha VL Teixeira HF Matte U Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells
description	Michelle Fraga1,2, Fernanda Bruxel1, Valeska Lizzi Lagranha2,3, Helder Ferreira Teixeira1, Ursula Matte2,31Post Graduation Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, 2Gene Therapy Center, Experimental Research Center, Hospital de Cl&iacute;nicas de Porto Alegre, 3Post Graduation Program in Genetics and Molecular Biology, Universidade Federal do Rio Grande do Sul, Porto Alegre, BrazilBackground: Cationic nanoemulsions have been recently considered as potential delivery systems for nucleic acids. This study reports the influence of phospholipids on the properties of cationic nanoemulsions/DNA plasmid complexes.Methods: Nanoemulsions composed of medium-chain triglycerides, stearylamine, egg lecithin or isolated phospholipids, ie, DSPC, DOPC, DSPE, or DOPE, glycerol, and water were prepared by spontaneous emulsification. Gene transfer to Hep G2 cells was analyzed using real-time polymerase chain reaction.Results: The procedure resulted in monodispersed nanoemulsions with a droplet size and zeta potential of approximately 250 nm and +50 mV, respectively. The complexation of cationic nanoemulsions with DNA plasmid, analyzed by agarose gel retardation assay, was complete when the complex was obtained at a charge ratio of &ge;1.0. In these conditions, the complexes were protected from enzymatic degradation by DNase I. The cytotoxicity of the complexes in Hep G2 cells, evaluated by MTT assay, showed that an increasing number of complexes led to progressive toxicity. Higher amounts of reporter DNA were detected for the formulation obtained with the DSPC phospholipid. Complexes containing DSPC and DSPE phospholipids, which have high phase transition temperatures, were less toxic in comparison with the formulations obtained with lecithin, DOPC, and DOPE.Conclusion: The results show the effect of the DNA/nanoemulsion complexes composition on the toxicity and transfection results.Keywords: plasmids, cationic nanoemulsions, phospholipids, physicochemical characterization, gene transfer, stearylamine
format	article
author	Fraga M Bruxel F Lagranha VL Teixeira HF Matte U
author_facet	Fraga M Bruxel F Lagranha VL Teixeira HF Matte U
author_sort	Fraga M
title	Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells
title_short	Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells
title_full	Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells
title_fullStr	Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells
title_full_unstemmed	Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells
title_sort	influence of phospholipid composition on cationic emulsions/dna complexes: physicochemical properties, cytotoxicity, and transfection on hep g2 cells
publisher	Dove Medical Press
publishDate	2011
url	https://doaj.org/article/36b64bf081f9498484b02dcd3bd1ec0d
work_keys_str_mv	AT fragam influenceofphospholipidcompositiononcationicemulsionsdnacomplexesphysicochemicalpropertiescytotoxicityandtransfectiononhepg2cells AT bruxelf influenceofphospholipidcompositiononcationicemulsionsdnacomplexesphysicochemicalpropertiescytotoxicityandtransfectiononhepg2cells AT lagranhavl influenceofphospholipidcompositiononcationicemulsionsdnacomplexesphysicochemicalpropertiescytotoxicityandtransfectiononhepg2cells AT teixeirahf influenceofphospholipidcompositiononcationicemulsionsdnacomplexesphysicochemicalpropertiescytotoxicityandtransfectiononhepg2cells AT matteu influenceofphospholipidcompositiononcationicemulsionsdnacomplexesphysicochemicalpropertiescytotoxicityandtransfectiononhepg2cells
_version_	1718400344639143936

Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells

Ejemplares similares