Cell-controlled and spatially arrayed gene delivery from fibrin hydrogels
Cell-controlled and spatially arrayed gene delivery from fibrin hydrogels
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DOI：
10.1016/j.biomaterials.2009.03.049
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发表时间：
2009-08-01
期刊：
影响因子：
14
通讯作者：
Andreadis, Stelios T.
中科院分区：
文献类型：
--
作者：
Lei, Pedro;Padmashali, Roshan M.;Andreadis, Stelios T.
我们研究了纤维蛋白介导的基因转移，在聚合过程中将pDNA嵌入水凝胶中，并使用两种基因转染模式将细胞放置在水凝胶表面(2D转染法)或水凝胶内(3D转染法)。利用这一模型，我们发现细胞转导强烈依赖于局部细胞-PDNA微环境，如2D与3D环境、靶细胞类型和密度以及纤维蛋白原和PDNA浓度。当细胞被包埋在纤维蛋白基质中时，脂联胺诱导的细胞死亡显著减少，尤其是在低靶细胞密度时。纤溶抑制剂的加入以一种剂量依赖的方式减少了基因转移，这表明纤维蛋白降解可能是有效基因转移所必需的。我们还提供了概念证明，纤维蛋白介导的基因转移可以用于空间定位的基因传递，这是细胞转染微阵列所必需的。当含有脂复合体的水凝胶以阵列形式被发现时，基因转移严格限制在含有PDNA的纤维蛋白斑点上，相邻位置之间没有交叉污染。总之，我们的数据表明，纤维蛋白可能被用作一种生物材料，以一种有效的、细胞控制的和空间定位的方式传递基因，在体外或体内具有潜在的应用前景。(C)2009爱思唯尔有限公司。保留所有权利。
We investigated fibrin-mediated gene transfer by embedding pDNA within the hydrogel during polymerization and using two modes of gene transfection with cells placed either on the surface (2D transfection) or within the hydrogel (3D transfection). Using this model, we found that cell transfection depended strongly on the local cell-pDNA microenvironment as defined by the 2D vs. 3D context, target cell type and density, as well as fibrinogen and pDNA concentrations. When cells were embedded within the fibrin matrix lipofectamine-induced cell death decreased significantly, especially at low target cell density. Addition of fibrinolytic inhibitors decreased gene transfer in a dose-dependent manner, suggesting that fibrin degradation may be necessary for efficient gene transfer. We also provided proof-of-concept that fibrin-mediated gene transfer can be used for spatially localized gene delivery, which is required in cell-transfection microarrays. When lipoplex-containing hydrogels were spotted in an array format gene transfer was strictly confined to pDNA-containing fibrin spots with no cross-contamination between neighboring sites. Collectively, our data suggest that fibrin may be used as a biomaterial to deliver genes in an efficient, cell-controlled and spatially localized manner for potential applications in vitro or in vivo. (C) 2009 Elsevier Ltd. All rights reserved.