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首页> 外文期刊>Acta biomaterialia >Differential uptake of DNA-poly(ethylenimine) polyplexes in cells cultured on collagen and fibronectin surfaces.
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Differential uptake of DNA-poly(ethylenimine) polyplexes in cells cultured on collagen and fibronectin surfaces.

机译：在胶原蛋白和纤连蛋白表面上培养的细胞中DNA-聚乙烯亚胺多聚体的差异摄取。

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摘要

Genetically modified bone marrow-derived mesenchymal stem cells (MSCs) have proven to be efficient cell carriers for local or systemic delivery of therapeutics as well as growth factors to augment tissue formation. However, efficient non-viral gene transfer to these cells is limiting their applicability. Although most studies have focused on designing more efficient condensation agents for DNA, our focus in this manuscript is to study the role of two extracellular matrix (ECM) proteins, collagen I (Col I) and fibronectin (Fn), on the ability of MSCs to become transfected. Here we report that plating MSCs on Col I-coated surfaces inhibits transfection, while plating MSCs on Fn-coated surfaces enhances transfection. The mechanism by which these ECM proteins affect non-viral gene transfer involves the endocytosis pathway used for polyplex uptake and intracellular tension. We found that Fn promoted internalization through clathrin-mediated endocytosis and that this pathway resulted in more efficient transfection than caveolae-mediated endocytosis and macropinocytosis. Further, the disruption of actin-myosin interactions resulted in an enhancement of gene transfer for cells plated on Fn-coated surfaces, but not for cells plated on Col I. We believe that the cellular microenvironment can be engineered to enhance the ability of cells to become transfected and that through understanding the mechanisms by which the ECM affects non-viral gene transfer better materials and transfection protocols can be realized.
机译：基因修饰的骨髓间充质干细胞（MSC）已被证明是有效的细胞载体，可局部或全身递送治疗药物以及生长因子以增强组织形成。但是，有效的非病毒基因转移到这些细胞限制了它们的适用性。尽管大多数研究都集中在设计更有效的DNA缩合剂上，但本文的重点是研究两种细胞外基质（ECM）蛋白胶原I（Col I）和纤连蛋白（Fn）对MSCs的作用。被转染。在这里，我们报道在Col I包被的表面上电镀MSC可以抑制转染，而在Fn包被的表面上电镀MSC可以提高转染。这些ECM蛋白影响非病毒基因转移的机制涉及用于多聚体摄取和细胞内张力的内吞途径。我们发现Fn通过网格蛋白介导的内吞作用促进内在化，并且该途径比小窝介导的内吞作用和巨胞饮作用导致更有效的转染。此外，肌动蛋白-肌球蛋白相互作用的破坏导致铺在Fn涂层表面的细胞的基因转移增强，但铺在Col I上的细胞却没有增强。我们相信可以改造细胞微环境，以增强细胞的能力。通过了解ECM影响非病毒基因转移的机制，可以实现更好的材料和转染方案。

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