Department of Cardiology, Cardiovascular Center, Korea University Anam Hospital, 145, Anam-ro, Seongbuk-gu, Seoul 02841, Republic of Korea.
School of Biomedical Engineering, College of Health Science, Korea University, 145, Anam-ro, Seongbuk-gu, Seoul 02841, Republic of Korea.
Acta Biomater. 2018 Jan;65:272-282. doi: 10.1016/j.actbio.2017.10.026. Epub 2017 Oct 14.
Nanotopography plays a pivotal role in the regulation of cellular responses. Nonetheless, little is known about how the gradient size of nanostructural stimuli alters the responses of endothelial progenitor cells without chemical factors. Herein, the fabrication of gradient nanopattern plates intended to mimic microenvironment nanotopography is described. The gradient nanopattern plates consist of nanopillars of increasing diameter ranges [120-200 nm (GP 120/200), 200-280 nm (GP 200/280), and 280-360 nm (GP 280/360)] that were used to screen the responses of human endothelial colony-forming cells (hECFCs). Nanopillars with a smaller nanopillar diameter caused the cell area and perimeter of hECFCs to decrease and their filopodial outgrowth to increase. The structure of vinculin (a focal adhesion marker in hECFCs) was also modulated by nanostructural stimuli of the gradient nanopattern plates. Moreover, Rho-associated protein kinase (ROCK) gene expression was significantly higher in hECFCs cultured on GP 120/200 than in those on flat plates (no nanopillars), and ROCK suppression impaired the nanostructural-stimuli-induced vinculin assembly. These results suggest that the gradient nanopattern plates generate size-specific nanostructural stimuli suitable for manipulation of the response of hECFCs, in a process dependent on ROCK signaling. This is the first evidence of size-specific nanostructure-sensing behavior of hECFCs.
Nano feature surfaces are of growing interest as materials for a controlled response of various cells. In this study, we successfully fabricated gradient nanopattern plates to manipulate the response of blood-derived hECFCs without any chemical stimulation. Interestingly, we find that the sensitive nanopillar size for manipulation of hECFCs is range between 120 nm and 200 nm, which decreased the area and increased the filopodial outgrowth of hECFCs. Furthermore, we only modulate the nanopillar size to increase ROCK expression can be an attractive method for modulating the cytoskeletal integrity and focal adhesion of hECFCs.
纳米形貌在调节细胞反应中起着关键作用。然而，对于没有化学因素的情况下，纳米结构刺激的梯度大小如何改变内皮祖细胞的反应，人们知之甚少。本文描述了旨在模拟微环境纳米形貌的梯度纳米图案板的制造。梯度纳米图案板由直径范围逐渐增大的纳米柱组成[120-200nm(GP 120/200)、200-280nm(GP 200/280)和 280-360nm(GP 280/360)]，用于筛选人内皮集落形成细胞(hECFC)的反应。纳米柱的纳米柱直径越小，hECFC 的细胞面积和周长就越小，丝状伪足的生长就越多。粘着斑蛋白(人内皮集落形成细胞中的粘着斑标志物)的结构也被梯度纳米图案板的纳米结构刺激所调节。此外，在 GP 120/200 上培养的 hECFC 的 Rho 相关蛋白激酶(ROCK)基因表达明显高于在平板(无纳米柱)上培养的 hECFC，并且 ROCK 抑制会损害纳米结构刺激诱导的粘着斑组装。这些结果表明，梯度纳米图案板产生适合于操纵 hECFC 反应的特定尺寸的纳米结构刺激，该过程依赖于 ROCK 信号。这是 hECFC 具有特定尺寸的纳米结构感知行为的第一个证据。
纳米特征表面作为控制各种细胞反应的材料越来越受到关注。在这项研究中，我们成功地制造了梯度纳米图案板，无需任何化学刺激即可操纵血液来源的 hECFC 的反应。有趣的是，我们发现用于操纵 hECFC 的敏感纳米柱尺寸范围在 120nm 和 200nm 之间，这降低了 hECFC 的面积并增加了丝状伪足的生长。此外，我们仅调节纳米柱尺寸以增加 ROCK 表达可以成为调节 hECFC 细胞骨架完整性和粘着斑的一种有吸引力的方法。
