Optimization of PEI/VEGF DNA polyplexes for potenital delivery from tissue engineering scaffolds

中国地球物理学会年刊 Pub Date : 2002-12-01 DOI:10.1109/IEMBS.2002.1136951

J. H. van Zanten, Y. Har-el, J. Hanes

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Abstract

Non-viral vector formation by DNA complexation with cationic condensing agents is a self-assembly process driven primarily by electrostatic interactions and counterion release. DNA complexation kinetics influence three physical parameters that have a direct effect on gene delivery and expression efficiency: DNA complex geometric size, surface charge and density. We demonstrate the utility of time resolved multiangle laser light scattering (TR-MALLS) for probing polyethylenimine (PEI) based polyplex formation kinetics with plasmid DNA. Our studies utilize plasmid DNA coding for VEGF, which may be used to enhance blood vessel in-growth into cell seeded polymeric scaffolds used in tissue engineering applications. We determined PEI/DNA complex size and density in real time and monitored vector stability in various liquid formulations. Parameters such as PEI molecular weight, N/P ratio and solution pH and ionic strength were investigated systematically. The ability to accurately measure polyplex size and density may lead to improvements in the design and control of non-viral gene delivery vectors and facilitate the determination of optimal formulations.

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PEI/VEGF DNA复合物的优化，用于组织工程支架的潜在递送

DNA与阳离子凝聚剂络合形成的非病毒载体是一个主要由静电相互作用和反离子释放驱动的自组装过程。DNA络合动力学影响三个对基因传递和表达效率有直接影响的物理参数:DNA复合物的几何尺寸、表面电荷和密度。我们展示了时间分辨多角度激光散射(tr - mall)用于探测基于聚乙烯亚胺(PEI)的多聚体形成动力学与质粒DNA的效用。我们的研究利用质粒DNA编码VEGF，可用于促进血管生长成细胞种子聚合物支架，用于组织工程应用。我们实时测定了PEI/DNA复合物的大小和密度，并监测了载体在各种液体配方中的稳定性。系统地考察了PEI的分子量、N/P比、溶液pH和离子强度等参数。精确测量多聚体大小和密度的能力可能导致非病毒基因传递载体的设计和控制的改进，并有助于确定最佳配方。

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中国地球物理学会年刊

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