Combining polyethylenimine and Fe(III) for mediating pDNA transfection.

Autor:	Jorge AF; Department of Chemistry, University of Coimbra, Rua Larga, Coimbra 3004-535, Portugal. Electronic address: andreiaj@qui.uc.pt., Röder R; Pharmaceutical Biotechnology, Center for System-based Drug Research, and Center for Nanoscience (CeNS), Ludwig-Maximilians-Universität München, Munich D-81377, Germany., Kos P; Pharmaceutical Biotechnology, Center for System-based Drug Research, and Center for Nanoscience (CeNS), Ludwig-Maximilians-Universität München, Munich D-81377, Germany., Dias RS; Biophysics and Medical Technology, Department of Physics, Norwegian University of Science and Technology, Trondheim 7491, Norway., Wagner E; Pharmaceutical Biotechnology, Center for System-based Drug Research, and Center for Nanoscience (CeNS), Ludwig-Maximilians-Universität München, Munich D-81377, Germany., Pais AA; Department of Chemistry, University of Coimbra, Rua Larga, Coimbra 3004-535, Portugal.
Jazyk:	angličtina
Zdroj:	Biochimica et biophysica acta [Biochim Biophys Acta] 2015 Jun; Vol. 1850 (6), pp. 1325-35. Date of Electronic Publication: 2015 Feb 19.
DOI:	10.1016/j.bbagen.2015.02.007
Abstrakt:	Background: The potential use of Fe(III) ions in biomedical applications may predict the interest of its combination with pDNA-PEI polyplexes. The present work aims at assessing the impact of this metal on pDNA complex properties. Methods: Variations in the formation of complexes were imposed by using two types of biological buffers at different salt conditions. The incorporation of pDNA in complexes was characterised by gel electrophoresis and dynamic light scattering. Transfection efficiency and cytotoxicity were evaluated in HeLa and HUH-7 cell lines, supported by flow cytometry assays. Results: Fe(III) enhances pDNA incorporation in the complex, irrespective of the buffer used. Transfection studies reveal that the addition of Fe(III) to complexes at low ionic strength reduces gene transfection, while those prepared under high salt content do not affect or, in a specific case, increase gene transfection up to 5 times. This increase may be a consequence of a favoured interaction of polyplexes with cell membrane and uptake. At low salt conditions, results attained with chloroquine indicate that the metal may inhibit polyplex endosomal escape. A reduction on the amount of PEI (N/P 5) formed at intermediary ionic strength, complemented by Fe(III), reduces the size of complexes while maintaining a transfection efficiency similar to that obtained to N/P 6. Conclusions: Fe(III) emerges as a good supporting condensing agent to modulate pDNA-PEI properties, including condensation, size and cytotoxicity, without a large penalty on gene transfection. General Significance: This study highlights important aspects that govern pDNA transfection and elucidates the benefits of incorporating the versatile Fe(III) in a gene delivery system. (Copyright © 2015 Elsevier B.V. All rights reserved.)
Databáze:	MEDLINE
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