Department of Biomedical Engineering, Duke University, 136 Hudson Hall, Box 90281, Durham, NC 27708, USA.
Biomaterials. 2011 May;32(14):3611-9. doi: 10.1016/j.biomaterials.2011.01.040. Epub 2011 Feb 21.
Optimization of nonviral gene delivery typically focuses on the design of particulate carriers that are endowed with desirable membrane targeting, internalization, and endosomal escape properties. Topographical control of cell transfectability, however, remains a largely unexplored parameter. Emerging literature has highlighted the influence of cell-topography interactions on modulation of many cell phenotypes, including protein expression and cytoskeletal behaviors implicated in endocytosis. Using high-throughput screening of primary human dermal fibroblasts cultured on a combinatorial library of microscale topographies, we have demonstrated an improvement in nonviral transfection efficiency for cells cultured on dense micropit patterns compared to smooth substrates, as verified with flow cytometry. A 25% increase in GFP(+) cells was observed independent of proliferation rate, accompanied by SEM and confocal microscopy characterization to help explain the phenomenon qualitatively. This finding encourages researchers to investigate substrate topography as a new design consideration for the optimization of nonviral transfection systems.
优化非病毒基因传递通常集中在设计具有理想的膜靶向、内化和内涵体逃逸特性的颗粒载体上。然而,细胞转染能力的形貌控制仍然是一个很大程度上未被探索的参数。新兴文献强调了细胞形貌相互作用对许多细胞表型的调节作用,包括涉及胞吞作用的蛋白质表达和细胞骨架行为。通过对在组合微尺度形貌库上培养的原代人真皮成纤维细胞进行高通量筛选,我们已经证明与光滑基底相比,在密集微孔图案上培养的细胞的非病毒转染效率得到了提高,这一点通过流式细胞术得到了验证。观察到 GFP(+)细胞的增加了 25%,而与增殖率无关,同时还进行了 SEM 和共聚焦显微镜的表征,以帮助定性地解释这一现象。这一发现鼓励研究人员将基底形貌作为优化非病毒转染系统的新设计考虑因素。