INEB
INEB
TitleOsteogenic differentiation of mesenchymal stem cells using PAMAM dendrimers as gene delivery vectors
Publication TypeJournal Article
2009
AuthorsSantos, JL, Oramas, E, Pêgo, AP, Granja, PL, Tomás, H
JournalJournal of Controlled ReleaseJ. Control. Release
Volume134
Issue2
Pagination141 - 148
Date Published2009///
01683659 (ISSN)
alkaline phosphatase, amine, Amines, animal cell, Animals, article, beta galactosidase, beta-Galactosidase, Bone, Bone morphogenetic protein, bone morphogenetic protein 2, Bone Regeneration, Calcium, Cell culture, cell differentiation, Cell Line, cell lineage, cell survival, Cells, Cells, Cultured, controlled study, dendrimer, Dendrimers, embryo, enzyme activity, Flowcharting, gene expression, Gene transfer, Gene Transfer Techniques, gene vector, Genes, Reporter, genetic transfection, human, human cell, Humans, in vitro study, Kidney, male, Mesenchymal stem cell, Mesenchymal Stem Cells, nonhuman, Nonviral gene delivery, nonviral gene delivery system, Nucleic acids, osteoblast, osteocalcin, Osteogenesis, PAMAM dendrimers, phosphate, plasmid, Plasmids, polyamidoamine, Polyamines, priority journal, protein secretion, Proteins, qualitative analysis, quantitative analysis, rat, Rats, Rats, Wistar, reporter gene, staining
This paper reports the use of different generations of polyamidoamine (PAMAM) dendrimers for the in vitro transfection of mesenchymal stem cells (MSCs). A systematic study was carried out on the transfection efficiency achieved by the PAMAM dendrimers using a β-galactosidase reporter gene system. Transfection results were shown to be dependent upon the generation of dendrimers, the amine to phosphate group ratio and the cell passage number. In all cases, the transfection efficiency was very low. Nevertheless, it was hypothesized that a low transfection level could be sufficient to promote the in vitro differentiation of MSCs towards the osteoblastic lineage. To address this possibility, dendrimers carrying the human bone morphogenetic protein-2 (hBMP-2) gene-containing plasmid were used. All quantitative (alkaline phosphatase activity, osteocalcin secretion and calcium deposition) and qualitative (von Kossa staining) osteogenic markers were significantly stronger in transfected cells when compared to non-transfected ones. This study not only clearly demonstrates that a low transfection level can be sufficient for inducing in vitro differentiation of MSCs to the osteoblast phenotype but also highlights the importance of focusing research on the development of gene delivery vectors in the concrete application. © 2008 Elsevier B.V. All rights reserved.
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