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首页> 外文期刊>Acta biomaterialia >Patterned transgene expression in multiple-channel bridges after spinal cord injury.
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Patterned transgene expression in multiple-channel bridges after spinal cord injury.

机译：脊髓损伤后多通道桥中模式化的转基因表达。

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摘要

Patterning of gene delivery on sub-millimeter length scales within tissue engineering scaffolds is fundamental to recreating the complex architectures of tissues. Surface-mediated delivery of lipoplexes mixed with fibronectin was investigated to pattern vectors within 250 microm channels in poly(lactide-co-glycolide) (PLG) bridges. Initial studies performed in vitro on PLG surfaces indicated that a DNA density of 0.07 microg mm(-2) inside each channel with a weight ratio of DNA to fibronectin of 1:20 maximized the number of transfected cells and the levels of transgene expression. Patterned vectors encoding for nerve growth factor (NGF) resulted in localized neurite extension within the channel. Translation to three-dimensional multiple-channel bridges enabled patterned transfection of different vectors throughout the channels for DNA:fibronectin ratios of 1:4 and multiple DNA depositions, with a large increase of neural cell bodies and neurite extension for delivery of DNA encoding for NGF. In vivo, the immobilization of non-viral vectors within the channels resulted in localized transfection within the pore structure of the bridge immediately around the channels of the bridge containing DNA. This surface immobilization strategy enables patterned gene delivery in vitro and in vivo on length scales of hundreds of microns and may find utility in strategies aimed at regenerating tissues with complex architectures.
机译：组织工程支架中亚毫米长度尺度的基因传递模式是重建复杂组织结构的基础。研究了与纤连蛋白混合的脂质复合物的表面介导递送，以在聚（丙交酯-乙交酯）（PLG）桥的250微米通道内对载体进行模式化。在PLG表面上进行的体外初步研究表明，每个通道内的DNA密度为0.07 microg mm（-2），DNA与纤连蛋白的重量比为1:20，可最大化转染细胞的数量和转基因表达的水平。编码神经生长因子（NGF）的模式化载体导致通道内局部神经突延伸。通过翻译成三维多通道桥，可以在整个通道中以1：4的DNA：纤连蛋白比率和多个DNA沉积进行不同载体的模式转染，并大量增加神经细胞体和神经突的延伸，以递送编码NGF的DNA 。在体内，非病毒载体在通道内的固定化导致在紧邻包含DNA的桥的通道周围的桥的孔结构内局部转染。这种表面固定策略可以在体外和体内以数百微米的长度尺度进行模式化的基因递送，并且可以在旨在再生具有复杂结构的组织的策略中找到实用性。

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