The enhancement of bone regeneration by gene activated matrix encoding for platelet derived growth factor.
The enhancement of bone regeneration by gene activated matrix encoding for platelet derived growth factor.
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DOI：
10.1016/j.biomaterials.2013.10.021
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发表时间：
2014-01
期刊：
影响因子：
14
通讯作者：
Salem, Aliasger K.
中科院分区：
文献类型：
--
作者：
Elangovan, Satheesh;D'Mello, Sheetal R.;Hong, Liu;Ross, Ryan D.;Allamargot, Chantal;Dawson, Deborah V.;Stanford, Clark M.;Johnson, Georgia K.;Sumner, D. Rick;Salem, Aliasger K.
关键词：
非病毒载体介导的基因治疗是克服蛋白质递送生长因子缺陷的有效途径。本研究的目的是开发和测试一种用于骨再生的非病毒基因递送系统，该系统利用胶原支架递送聚乙烯亚胺（PEI）-质粒DNA（pDNA）[编码血小板衍生生长因子-B（PDGF-B）]复合物。PEI-pPDGF-B复合物以胺（N）与磷酸盐（P）的比例为10制备，并表征尺寸、表面电荷和体外细胞毒性以及在人骨髓基质细胞（BMSC）中的转染功效。使用显微镜技术在体外评价复合负载的胶原支架对细胞附着和募集的影响。在Fisher 344大鼠中，在5 mm直径的临界大小的颅骨缺损中评估基因递送系统的体内再生能力。复合物的尺寸约为100 nm，具有正表面电荷。以N/P比为10制备的复合物显示出低细胞毒性，如通过细胞活力测定所评估的。共聚焦显微镜显示，与空支架相比，复合负载胶原支架上的BMSCs显着增殖。体内研究显示，与空缺损或空支架相比，植入4周后，用复合物活化支架治疗的颅骨缺损中的新骨体积/总体积（BV/TV）%显著更高（分别高14倍和44倍）。总之，这些研究结果表明，非病毒PDGF-B基因激活支架对骨再生有效，是一种有吸引力的基因递送系统，具有显著的临床转化潜力。
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 microscopic 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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