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Title: Enhanced intracellular uptake and endocytic pathway selection mediated by hemocompatible ornithine grafted chitosan polycation for gene delivery
Authors: Alex, SM
Sharma, CP
Keywords: Biophysics; Chemistry; Materials Science
Issue Date: 2014
Citation: 122 ,;792-800
Abstract: Nanotechnology is adopted in gene therapy research to create gene vectors that will facilitate gene transfer to cells with utmost efficacy and safety. For vector design, polymers are the preferred nonviral colloidal systems as they are feasible for any chemical modifications. In this study, chitosan, a versatile biopolymer has been subjected to chemical conjugation with the amino acid ornithine to generate chitosan-ornithine conjugate (CON) for gene delivery. With the help of FTIR and H-1 NMR spectra the chemical composition of the chitosan derivative was confirmed. Buffering capacity was found enhanced with the synthesised chitosan derivative when compared to the parent unmodified chitosan. The cationic derivative formed nanoparticles when mixed with negatively charged DNA. The nanoparticles showed good DNA retardation ability in agarose gel electrophoresis and sizes were ascertained by DLS and TEM observations. The derivative on interaction with blood plasma showed negligible protein adsorption and did not cause either hemolysis or RBC aggregation in blood. In vitro cell culture also revealed the CON derivative to be nontoxic to cells and capable of transfection with an explicit increase in cellular uptake of nanopartides. An uptake study in the presence of endocytosis inhibitors indicated the specific pathway used for cell entry. The results revealed that the clathrin mediated pathway and dynamin played a role in the internalisation of these specific nanoparticles. (C) 2014 Published by Elsevier B.V.
URI: 10.1016/j.colsurfb.2014.08.023
Appears in Collections:Journal Articles

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