INEB
INEB
TitleCationic liposome-DNA complexes as gene delivery vectors: Development and behaviour towards bone-like cells
Publication TypeJournal Article
2009
AuthorsOliveira, AC, Ferraz, MP, Monteiro, FJ, Simões, S
JournalActa BiomaterialiaActa Biomater.
Volume5
Issue6
Pagination2142 - 2151
Date Published2009///
17427061 (ISSN)
1,2 dioleoyl 3 trimethylammoniopropane, animal cell, Animals, article, bone cell, bone development, bone tissue, Cationic liposomes, Cell Line, cell type, complex formation, controlled study, Cytotoxicity, DNA, drug mechanism, gene expression regulation, Gene therapy, Gene transfer, genetic transfection, Genetic Vectors, human, human cell, internalization, lipoplex, liposome, Liposomes, materials testing, Mice, nonhuman, nonviral gene delivery system, osteoblast, Osteoblasts, Osteogenesis, Particle Size, plasmid, priority journal, Transfection, transferrin, Zeta potential
Modulation of the biological pathways responsible for fracture repair and osteogenisis may accelerate regeneration. Gene therapy is an alternative method for the release of osteogenisis-stimulating proteins into tissues. The development of vectors for gene release is still a problem in terms of ethics and techniques. In this work we evaluated whether cationic liposomes constitute a valuable strategy for the release of genetic material into bone tissue cells as non-viral vectors. Liposomes were prepared with 1,2-dioleoyl-3-trimethylammonium propane (DOTAP)-2-dioleoyl-sn-glycero-3-phosphatidylethanolamine and DOTAP-cholesterol, and characterized according to their size, zeta potential, DNA protection capacity and cytotoxicity. Transfection studies were also carried out using pCMVβ-gal plasmid in two osteoblastic cell lines (MG63 and MC3T3-E1) and in the 294T line, varying the charge ratio and the applied DNA dose. Inclusion of transferrin to increase the expression was also tested. The results suggest that there is great dependency between the transfection activity and the lipid formulation, the charge ratios of the complexes, the applied DNA dose and the cell type. There were even some differences concerning both osteoblastic lines under study. The cells of the MC3T3-E1 line present greater expression levels than the cells of the MG-63 line. The conjugation of the transferrin with the complexes contributes to the increase in transfection levels, possibly due to an increase in internalization of complexes. It is thus a good strategy for inducing the expression of specific genes in osteoblast-like cells. © 2009 Acta Materialia Inc.
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