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- Title
Non-viral-mediated gene therapy approaches for bone repair.
- Authors
Winn, S. R.; Chen, J. C.; Gong, X.; Bartholomew, S. V.; Shreenivas, S.; Ozaki, W.
- Abstract
<bold>Objectives: </bold>Bone repair strategies continue to be developed for alternatives to autografting, allogeneic implants of banked bone, and other bone substitutes. Efforts have included the delivery of potent growth and/or differentiation factors and the use of gene therapy. For bone regeneration, gene therapy is the delivery, uptake and expression of DNA that has been localized to a wound bed. The objective of the current study is to investigate methods to enhance non-viral-mediated means of gene uptake and expression for use in bone regeneration.<bold>Methods: </bold>Several types of DNA-polymer complexes, either applied directly to baby hamster kidney (BHK) cells, or released from a porous, resorbable gene-activated matrix (GAM), were evaluated in vitro for their ability to transfect cells with a circular plasmid DNA construct expressing green fluorescent protein. Complexes included conjugates containing a lipophilic reagent, liposomes, poly-ethyl-oxazoline, and poly-ethyleneimine (PEI). Data were subjected to analysis of variance and Fisher's protected least significant difference for multiple comparisons with significance established at p < 0.05.<bold>Results: </bold>Transfection efficiencies of the liposome and PEI complexes improved in vitro when released from resorbable GAMs. The lipophilic reagent FuGene 6 demonstrated abundant uptake and expression in the initial 1- and 2-day evaluation periods. In contrast, the DNA-liposome and PEI GAM complexes demonstrated a sustained release, uptake and expression by the BHK cells at the 2-, 4-, and 7-day, and 4- and 7-day evaluation intervals, respectively.<bold>Conclusion: </bold>GAM technology appears to improve the functional stability and release duration of incorporated DNA-polymer complexes in the present in vitro studies. The ongoing objective of our research is to develop a localized treatment to improve the uptake and expression of plasmid DNA by non-viral-mediated gene therapy.
- Subjects
GENE therapy; DNA; LIPOSOMES; REGENERATION (Biology); GENE transfection; CELLS
- Publication
Orthodontics & Craniofacial Research, 2005, Vol 8, Issue 3, p183
- ISSN
1601-6335
- Publication type
journal article
- DOI
10.1111/j.1601-6343.2005.00332.x