The enhancement of bone regeneration by gene activated matrix encoding for platelet derived growth factor |
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Authors: | Satheesh Elangovan Sheetal R D'Mello Liu Hong Ryan D Ross Chantal Allamargot Deborah V Dawson Clark M Stanford Georgia K Johnson D Rick Sumner Aliasger K Salem |
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Institution: | 1. Department of Periodontics, University of Iowa College of Dentistry, Iowa City, IA, USA;2. Division of Pharmaceutics and Translational Therapeutics, University of Iowa College of Pharmacy, Iowa City, IA, USA;3. Department of Prosthodontics, University of Iowa College of Dentistry, Iowa City, IA, USA;4. Department of Anatomy and Cell Biology, Rush Medical College, Chicago, IL, USA;5. Central Microscopy Research Facility, University of Iowa, Iowa City, IA, USA;6. Division of Biostatistics and Research Design, College of Dentistry, Depts. of Pediatric Dentistry & Biostatistics and the Interdisciplinary Programs in Genetics and in Informatics, University of Iowa, Iowa City, IA, USA |
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Abstract: | Gene therapy using non-viral vectors that are safe and efficient in transfecting target cells is an effective approach to overcome the shortcomings of protein delivery of growth factors. The objective of this study was to develop and test a non-viral gene delivery system for bone regeneration utilizing a collagen scaffold to deliver polyethylenimine (PEI)-plasmid DNA (pDNA) encoding platelet derived growth factor-B (PDGF-B)] complexes. The PEI-pPDGF-B complexes were fabricated at amine (N) to phosphate (P) ratio of 10 and characterized for size, surface charge, and in vitro cytotoxicity and transfection efficacy in human bone marrow stromal cells (BMSCs). The influence of the complex-loaded collagen scaffold on cellular attachment and recruitment was evaluated in vitro using microscopy techniques. The in vivo regenerative capacity of the gene delivery system was assessed in 5 mm diameter critical-sized calvarial defects in Fisher 344 rats. The complexes were ∼100 nm in size with a positive surface charge. Complexes prepared at an N/P ratio of 10 displayed low cytotoxicity as assessed by a cell viability assay. Confocal microscopy revealed significant proliferation of BMSCs on complex-loaded collagen scaffolds compared to empty scaffolds. In vivo studies showed significantly higher new bone volume/total volume (BV/TV) % in calvarial defects treated with the complex-activated scaffolds following 4 weeks of implantation (14- and 44-fold higher) when compared to empty defects or empty scaffolds, respectively. Together, these findings suggest that non-viral PDGF-B gene-activated scaffolds are effective for bone regeneration and are an attractive gene delivery system with significant potential for clinical translation. |
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Keywords: | Plasmid DNA Polyethylenimine Gene delivery Scaffold Platelet derived growth factor Bone regeneration |
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