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dc.contributor.authorAlexander, Jennifer C.
dc.contributor.authorBrowne, Shane
dc.contributor.authorPandit, Abhay
dc.contributor.authorRochev, Yury
dc.date.accessioned2018-09-20T15:59:21Z
dc.date.available2018-09-20T15:59:21Z
dc.date.issued2013-06-03
dc.identifier.citationAlexander, Jennifer C. Browne, Shane; Pandit, Abhay; Rochev, Yury (2013). Biomaterial constructs for delivery of multiple therapeutic genes: a spatiotemporal evaluation of efficacy using molecular beacons. PLoS ONE 8 (6),
dc.identifier.issn1932-6203
dc.identifier.urihttp://hdl.handle.net/10379/10184
dc.description.abstractGene therapy is emerging as a potential therapeutic approach for cardiovascular pathogenesis. An appropriate therapy may require multiple genes to enhance therapeutic outcome by modulating inflammatory response and angiogenesis in a controlled and time-dependent manner. Thus, the aim of this research was to assess the spatiotemporal efficacy of a dualgene therapy model based on 3D collagen scaffolds loaded with the therapeutic genes interleukin 10 (IL-10), a potent anti-inflammatory cytokine, and endothelial nitric oxide synthase (eNOS), a promoter of angiogenesis. A collagen-based scaffold loaded with plasmid IL-10 polyplexes and plasmid eNOS polyplexes encapsulated into microspheres was used to transfect HUVECs and HMSCs cells. The therapeutic efficacy of the system was monitored at 2, 7 and 14 days for eNOS and IL-10 mRNA expression using RT-PCR and live cell imaging molecular beacon technology. The dual gene releasing collagen-based scaffold provided both sustained and delayed release of functional polyplexes in vitro over a 14 day period which was corroborated with variation in expression levels seen using RT-PCR and MB imaging. Maximum fold increases in IL-10 mRNA and eNOS mRNA expression levels occurred at day 7 in HMSCs and HUVECs. However, IL-10 mRNA expression levels seemed dependent on frequency of media changes and/or ease of transfection of the cell type. It was demonstrated that molecular beacons are able to monitor changes in mRNA levels at various time points, in the presence of a 3D scaffolding gene carrier system and the results complemented those of RT-PCR.
dc.publisherPublic Library of Science (PLoS)
dc.relation.ispartofPLoS ONE
dc.rightsAttribution-NonCommercial-NoDerivs 3.0 Ireland
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/3.0/ie/
dc.subjectmesenchymal stem-cells
dc.subjectnitric-oxide synthase
dc.subjectmessenger-rna
dc.subjectliving cells
dc.subjectinterleukin-10
dc.subjectexpression
dc.subjecthybridization
dc.subjectprediction
dc.subjectdisease
dc.subjectgrowth
dc.titleBiomaterial constructs for delivery of multiple therapeutic genes: a spatiotemporal evaluation of efficacy using molecular beacons
dc.typeArticle
dc.identifier.doi10.1371/journal.pone.0065749
dc.local.publishedsourcehttps://doi.org/10.1371/journal.pone.0065749
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Attribution-NonCommercial-NoDerivs 3.0 Ireland
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivs 3.0 Ireland