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Dadun > Depósito Académico > Facultad de Farmacia > Departamento de Farmacia y Tecnología Farmacéutica > DA - Farmacia - Tecnología Farmacéutica - Artículos de revista >

New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles
Authors: Centelles, M.N. (Miguel N.)
Qian, C. (Cheng)
Campanero, M.A. (Miguel Angel)
Irache, J.M. (Juan Manuel)
Keywords: Chitosan
Nanoparticles
Gene delivery
Hydrodynamics-based procedure
Bioluminescence
Intestine
Issue Date: 2008
Publisher: Dove Medical Press Ltd.
Publisher version: http://www.dovepress.com/articles.php?article_id=2396. DOI: 10.2147/IJN.S3445
ISSN: 1176-9114
Citation: Centelles MN, Qian C, Campanero MA, Irache JM. New methodologies to characterize the effectiveness of the gene transfer mediated by DNA-chitosan nanoparticles. Int J Nanomedicine 2008;3(4):451-460.
Abstract
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 signifi cant infl uence 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 profi le, 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 quantifi able 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.
Permanent link: http://hdl.handle.net/10171/17090
Appears in Collections:DA - CUN - Farmacología clínica - Artículos de revista
DA - Farmacia - Tecnología Farmacéutica - Artículos de revista

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