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Nanoparticle uptake and gene transfer efficiency for MSCs on chitosan and chitosan-hyaluronan substrates

Paper ID Volume ID Publish Year Pages File Format Full-Text
6943 525 2012 12 PDF Available
Title
Nanoparticle uptake and gene transfer efficiency for MSCs on chitosan and chitosan-hyaluronan substrates
Abstract

Nanoparticles (NPs) are usually surface modified to increase endocytosis for applications in cellular imaging and gene delivery. The influence of cell culture substrates on endocytosis remains relatively unexplored. This study investigated the substrate-mediated effects on the uptake of NPs by mesenchymal stem cells (MSCs). Two types of NPs were employed, negatively charged paramagnetic iron oxide (Fe3O4) NPs (∼5 nm) and bare plasmid DNA pTRE-Tight-DsRED2 (3.3 kb, ∼5 nm), each of which were poorly endocytosed by the adipose-derived MSCs grown on tissue culture polystyrene (TCPS). When cells were cultured on chitosan or hyaluronan-modified chitosan (chitosan-HA) membranes, significant increases (>5-fold) in the intracellular uptake of Fe3O4 NPs as well as transfectability of plasmid DNA were demonstrated. The enhancement in transgene expression was more pronounced than that using the transfection agent. The beneficial effects were not caused by elevated proliferation or a change in the differentiation state of interacting MSCs. On chitosan and chitosan-HA, cells moved fast and formed spheroids. The cytoskeletal arrangement associated with the up-regulated RhoA activity during spheroid formation may have accounted for the increased endocytosis. Using different inhibitors, the endocytosis pathways were further clarified. Both Fe3O4 NPs and plasmid DNA were taken up primarily by clathrin-mediated endocytosis on chitosan (∼50%). The caveolae-mediated endocytosis on chitosan-HA was more evident (∼30–40%) than that on chitosan (<25%). For plasmid DNA but not Fe3O4 NPs, macropinocytosis also occurred on both substrates. Chitosan and chitosan-HA as cell culture substrates may activate different endocytic pathways of MSCs to increase NP internalization or plasmid transfection. The substrate-mediated endocytosis described here may represent a new and potentially attractive approach to facilitate stem cell labeling or to improve gene delivery efficiency without altering cell viability and differentiation.

Keywords
Nanoparticles (NPs); Mesenchymal stem cells (MSCs); Endocytosis; Gene transfection; Chitosan
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Nanoparticle uptake and gene transfer efficiency for MSCs on chitosan and chitosan-hyaluronan substrates
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 33, Issue 14, May 2012, Pages 3639–3650
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us