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Permanent link (DOI): https://doi.org/10.7939/R3QF8JR68

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Theses and Dissertations

In vivo gene delivery with non-viral carriers Open Access

Descriptions

Other title
Subject/Keyword
Gene delivery
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Rose, Laura C
Supervisor and department
Uludag, Hasan (Chemical Engineering)
Examining committee member and department
Unsworth, Larry (Chemical Engineering)
Rice, Kevin (Pharmaceutical Sciences and Experimental Therapeutics)
El-Bialy, Tarek (Dentistry)
Burrell, Robert (Biomedical Engineering)
Gorassini, Monica (Biomedical Engineering)
Adesida, Adetola (Surgery)
Department
Department of Biomedical Engineering
Specialization

Date accepted
2013-11-13T15:29:32Z
Graduation date
2014-06
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
Gene delivery is an emerging therapy for treatment of many different diseases and conditions, with the first therapy recently approved for clinical use. This thesis investigates non-viral carriers for gene delivery for bone regeneration, with an emphasis on the mechanisms of plasmid DNA (pDNA) delivery and methods to improve transfection. Polyethylenimine (PEI, 2 kDa) modified with linoleic acid (PEI-LA) was found to give transfection rates comparable to viral vectors both in vitro and in vivo. The PEI-LA/pDNA complexes were found to display a decreased transfection efficiency over time, but a gelatin coating was found to prevent this loss of transfection. The gelatin-coated particles also led to increased transfection in vivo, allowing lower doses of pDNA to be used. Finally, we developed a novel quantitative PCR (qPCR) method to detect pDNA bound in a polymer complex such that the pharmacokinetics could be investigated. The pDNA delivered without a polymeric carrier was degraded very rapidly, while pDNA from PEI and PEI-LA complexes were detectable for two and four weeks respectively. For polymeric complexes, the qPCR method was in good agreement with studies tracking fluorescently labelled pDNA. Similar to in vitro results, PEI complexes gave no gene expression while PEI-LA complexes gave gene expression for at least four weeks. Although no bone regeneration was observed following delivery of complexes, these studies provide crucial information on the non-viral gene delivery in vivo.
Language
English
DOI
doi:10.7939/R3QF8JR68
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication
L Rose, H Uludag. Realizing the potential of gene-based molecular medicines in bone repair. J Bone Miner Res 2013;28(11):2245-62.LC Rose, R Fitzsimmons, P Lee, R Krawetz, DE Rancourt, H Uludag. Effect of basic fibroblast growth factor in mouse embryonic stem cell culture and osteogenic differentiation. J Tissue Eng Regen Med 2013;7(5):371-82.LC Rose, C Kucharski, H Uludag. Protein expression following non-viral delivery of plasmid DNA coding for basic FGF and BMP-2 in a rat ectopic model. Biomaterials 2012;33(11):3363-74.L Rose, HM Aliabadi, H Uludag. Gelatin coating to stabilize the transfection ability of nucleic acid polyplexes. Acta Biomater 2013;9(7):7429-38.L Rose, P Mahdipoor, C Kucharski, H Uludag. Pharmacokinetics and transgene expression of implanted polyethylenimine-based pDNA complexes. Biomaterials Science 2013. BM-ART-08-2013-060200. In press.

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