New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles

Miguel N Centelles1, Cheng Qian2, Miguel A Campanero1, Juan M Irache11Centro Gal&eacute;nico, Departamento Farmacia y Tecnolog&iacute;a Farmac&eacute;utica, University of Navarra, Irunlarrea 1, 31080, Pamplona, Spain; 2Centro de Investigaci&oacute;n Medica Aplicada (C...

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Autores principales:	Miguel N Centelles, Cheng Qian, Miguel A Campanero, Juan M Irache
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2008
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/8d8a9960a4e848f98eb4f74a403d3219
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spelling	oai:doaj.org-article:8d8a9960a4e848f98eb4f74a403d32192021-12-02T02:10:27ZNew methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles1176-91141178-2013https://doaj.org/article/8d8a9960a4e848f98eb4f74a403d32192008-10-01T00:00:00Zhttp://www.dovepress.com/new-methodologies-to-characterize-the-effectiveness-of-the-gene-transf-a2396https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Miguel N Centelles1, Cheng Qian2, Miguel A Campanero1, Juan M Irache11Centro Gal&eacute;nico, Departamento Farmacia y Tecnolog&iacute;a Farmac&eacute;utica, University of Navarra, Irunlarrea 1, 31080, Pamplona, Spain; 2Centro de Investigaci&oacute;n Medica Aplicada (CIMA), Facultad de Medicina, Gene Therapy Unit, University of Navarra, 31008 Pamplona, SpainAbstract: In this work three DNA-chitosan nanoparticle formulations (Np), differing in the molecular weight (MW; 150 kDa, 400 kDa, and 600 kDa) of the polysaccharide, were prepared and administered by two different administration routes: the hydrodynamics-based procedure and the intraduodenal injection. After the hydrodynamic injection, DNA-chitosan nanoparticles were predominantly accumulated in the liver, where the transgene was expressed during at least 105 days. No significant influence of MW was observed on the levels of luciferase expression. The curves of bioluminescence versus time obtained using the charge-coupled device (CCD) camera were described and divided in three phases: (i) the initial phase, (ii) the sustained release step and (iii) the decline phase (promotor inactivation, immunological and physiological processes). From these curves, which describe the transgene expression profile, the behavior of the different formulations as gene delivery systems was characterized. Therefore, the following parameters such as Cmax (maximum level of detected bioluminescence), AUC (area under the bioluminescence-time curve) and MET (mean time of the transgene expression) were calculated. This approach offers the possibility of studying and comparing transgene expression kinetics among a wide variety of gene delivery systems. Finally, the intraduodenal administration of naked DNA permitted the gene transfer in a dose dependent manner quantifiable with the CCD camera within 3 days. Nevertheless, the same administration procedure of the three formulations did not improve the levels of transgene expression obtained with naked DNA. This fact could be explained by the rapid physiological turn-over of enterocytes and by the ability of chitosan nanoparticles to control the DNA release.Keywords: chitosan, nanoparticles, gene delivery, hydrodynamics-based procedure, bioluminescence, intestine Miguel N CentellesCheng QianMiguel A CampaneroJuan M IracheDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2008, Iss Issue 4, Pp 451-460 (2008)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Miguel N Centelles Cheng Qian Miguel A Campanero Juan M Irache New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles
description	Miguel N Centelles1, Cheng Qian2, Miguel A Campanero1, Juan M Irache11Centro Gal&eacute;nico, Departamento Farmacia y Tecnolog&iacute;a Farmac&eacute;utica, University of Navarra, Irunlarrea 1, 31080, Pamplona, Spain; 2Centro de Investigaci&oacute;n Medica Aplicada (CIMA), Facultad de Medicina, Gene Therapy Unit, University of Navarra, 31008 Pamplona, SpainAbstract: In this work three DNA-chitosan nanoparticle formulations (Np), differing in the molecular weight (MW; 150 kDa, 400 kDa, and 600 kDa) of the polysaccharide, were prepared and administered by two different administration routes: the hydrodynamics-based procedure and the intraduodenal injection. After the hydrodynamic injection, DNA-chitosan nanoparticles were predominantly accumulated in the liver, where the transgene was expressed during at least 105 days. No significant influence of MW was observed on the levels of luciferase expression. The curves of bioluminescence versus time obtained using the charge-coupled device (CCD) camera were described and divided in three phases: (i) the initial phase, (ii) the sustained release step and (iii) the decline phase (promotor inactivation, immunological and physiological processes). From these curves, which describe the transgene expression profile, the behavior of the different formulations as gene delivery systems was characterized. Therefore, the following parameters such as Cmax (maximum level of detected bioluminescence), AUC (area under the bioluminescence-time curve) and MET (mean time of the transgene expression) were calculated. This approach offers the possibility of studying and comparing transgene expression kinetics among a wide variety of gene delivery systems. Finally, the intraduodenal administration of naked DNA permitted the gene transfer in a dose dependent manner quantifiable with the CCD camera within 3 days. Nevertheless, the same administration procedure of the three formulations did not improve the levels of transgene expression obtained with naked DNA. This fact could be explained by the rapid physiological turn-over of enterocytes and by the ability of chitosan nanoparticles to control the DNA release.Keywords: chitosan, nanoparticles, gene delivery, hydrodynamics-based procedure, bioluminescence, intestine
format	article
author	Miguel N Centelles Cheng Qian Miguel A Campanero Juan M Irache
author_facet	Miguel N Centelles Cheng Qian Miguel A Campanero Juan M Irache
author_sort	Miguel N Centelles
title	New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles
title_short	New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles
title_full	New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles
title_fullStr	New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles
title_full_unstemmed	New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles
title_sort	new methodologies to characterize the effectiveness of the gene transfer mediated by dna-chitosan nanoparticles
publisher	Dove Medical Press
publishDate	2008
url	https://doaj.org/article/8d8a9960a4e848f98eb4f74a403d3219
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New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles

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