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Intracellular Trafficking and Decondensation Kinetics of Chitosan–pDNA Polyplexes

Marc Thibault, Surendra Nimesh, Marc Lavertu and Michael D. Buschmann

Article (2010)

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The transfection efficiency (TE) of chitosan-plasmid DNA (pDNA) polyplexes can be critically modulated by the polymer degree of deacetylation (DDA) and molecular weight (MW). This study was performed to test the hypothesis that the TE dependence on chitosan MW and DDA is related to the polyplex stability, hence their intracellular decondensation/unpacking kinetics. Major barriers to nonviral gene transfer were studied by image-based quantification. Although uptake increased with increased DDA, it did not appear to be a structure-dependent process affecting TE, nor was nuclear entry. Colocalization analysis showed that all chitosans trafficked through lysosomes with similar kinetics. Fluorescent resonant energy transfer (FRET) analysis revealed a distinct relationship between TE and polyplex dissociation rate. The most efficient chitosans showed an intermediate stability and a kinetics of dissociation, which occurred in synchrony with lysosomal escape. In contrast, a rapid dissociation before lysosomal escape was found for the inefficient low DDA chitosan whereas the highly stable and inefficient complex formed by a high MW and high DDA chitosan did not dissociate even after 24 hours. This study identified that the kinetics of decondensation in relation to lysosomal escape was a most critical structure-dependent process affecting the TE of chitosan polyplexes.

Uncontrolled Keywords

Cell Line; Chitosan; DNA; Flow Cytometry; Fluorescence Resonance Energy Transfer; Humans; Kinetics; Lysosomes; Molecular Weight; Plasmids; Transfection; DNA; Chitosan

Subjects: 1800 Chemical engineering > 1800 Chemical engineering
1900 Biomedical engineering > 1900 Biomedical engineering
Department: Department of Chemical Engineering
Institut de génie biomédical
Funders: CISH, CRSNG/NSERC, Fond de la recherche en santé du Québec, Canada Foundation for Innovation
PolyPublie URL: https://publications.polymtl.ca/3402/
Journal Title: Molecular Therapy (vol. 18, no. 10)
Publisher: Elsevier
DOI: 10.1038/mt.2010.143
Official URL: https://doi.org/10.1038/mt.2010.143
Date Deposited: 10 Jan 2019 14:20
Last Modified: 07 Nov 2023 11:18
Cite in APA 7: Thibault, M., Nimesh, S., Lavertu, M., & Buschmann, M. D. (2010). Intracellular Trafficking and Decondensation Kinetics of Chitosan–pDNA Polyplexes. Molecular Therapy, 18(10), 1787-1795. https://doi.org/10.1038/mt.2010.143


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