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- Title
535. Physicochemical and Transfection Properties of Cationic Hydroxyethylcellulose/DNA Nanoparticles.
- Authors
Fayazpour, Farzaneh; Lucas, Bart; Alvarez-Lorenzo, Carmen; Sanders, Niek; Demeester, Joseph; De Smedt, Stefaan
- Abstract
Purpose:Recently, cationic polysaccharides were investigated for DNA delivery purposes. PQ-4 and PQ-10 two types of cationic hydroxyethylcellulose were previously characterized in detail and already have applications in cosmetics and topic drug delivery devices. In this study we evaluate their DNA complexation and transfection properties. Their DNA binding and transfection properties were compared with the branched and linear polyethylenimie (PEI) which are widely investigated for DNA delivery purposes.Methods:Size distribution and surface charge properties of the polyplexes were determined by DLS and zeta potential measurements. The complexation of pDNA to the cationic polymers and dissociation of the polyplexes by polyanions (dextran sulfate or poly-L-aspartic acid) were studied by gelelectrophoresis and the picogreen fluorescence assay.Transfection efficiencies using Cos-7 cells were evaluated by using the SEAP assy. The cell viability towards the polyplexes was determined using EZ4U assay.Results:Gelelectrophoresis and fluorescence experiments indicated that PQ-4 and PQ-10 can bind and condense pDNA. Compared to PEI, the interaction between pDNA and cationic HEC seems to be stronger. Although, the gelelectrophoresis data showed that PQ-10 binds all the pDNA, the fluorescent quenching assay indicates that PQ-10 does not condense pDNA as tightly as the other cationic polymers, still allowing picrogreen to intercalate to a certain extent in the pDNA.PEI based polyplexes transfect significantly better than cationic cellulose/pdna complexes.The transfection efficiency of the cellulase pretreated PQ-10: compared to high molecular weight PQ-10, transfects better, however, still less efficient than BPEI.PQ-4 and PQ-10 based pDNA complexes show low toxicity as more than 90% of the cells remains alive. At higher N/P ratio the cationic cellulose/ pDNA dispersions seemed to be slightly more toxic which may be due to the presence of a higher amount of (free) cationic cellulose damaging the cellular membranes.Conclusion:It is hypothesized that due to the strong binding and the observed uncharged surface, PQ-4/pDNA polyplexes do not optimally interact with cellular membranes. Consequently they are not taken up by cells which explain why they do not transfect cells. Coupling of a ligand for receptor-mediated uptake might overcome the problems of the neutrally charged polyplexes.PQ-10 proves to be a cationic polymer with a rather surprising behavior: It binds pDNA strongly while the latter does not become fully condensed. The PQ-10/pDNA complexes used in this study did transfect cells to a certain extent, however, not as efficient of PEI based polyplexes. Tailoring of the nature and extent of cationic side chains on this type of cationic hydroxyethylcellulose may be promising to further enhance their DNA delivery properties.Molecular Therapy (2006) 13, S206–S206; doi: 10.1016/j.ymthe.2006.08.606
- Subjects
GENETIC transformation; DNA; BIOPOLYMERS; POLYSACCHARIDES; DRUG delivery systems
- Publication
Molecular Therapy, 2006, Vol 13, pS206
- ISSN
1525-0016
- Publication type
Article
- DOI
10.1016/j.ymthe.2006.08.606