The enhancement of bone regeneration by gene activated matrix encoding for platelet derived growth factor

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• 作者：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
• 关键词：Plasmid DNA ; Polyethylenimine ; Gene delivery ; Scaffold ; Platelet derived growth factor ; Bone regeneration
• 刊名：Biomaterials
• 出版年：January, 2014
• 年：2014
• 卷：35
• 期：2
• 页码：737-747
• 全文大小：2473 K

文摘

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聽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聽vitro using microscopy techniques. The in聽vivo regenerative capacity of the gene delivery system was assessed in 5聽mm diameter critical-sized calvarial defects in Fisher 344 rats. The complexes were 鈭?00聽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聽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.