Dhawan Udesh, Pan Hsu An, Lee Chia Hui, Chu Ying Hao, Huang Guewha Steven, Lin Yan Ren, Chen Wen Liang
Department of Materials Science and Engineering, National Chiao Tung University, Hsinchu, Taiwan, ROC.
Hokan Life Technology, Taichung, Taiwan, ROC.
PLoS One. 2016 Jun 30;11(6):e0158425. doi: 10.1371/journal.pone.0158425. eCollection 2016.
Nanotopological cues can be exploited to understand the nature of interactions between cells and their microenvironment to generate superior implant geometries. Nanosurface parameters which modulate the cell behavior and characteristics such as focal adhesions, cell morphology are not clearly understood. Here, we studied the role of different nanotopographic dimensions in modulating the cell behavior, characteristics and ultimately the cell fate and accordingly, a methodology to improve implant surface geometry is proposed. Tantalum oxide nanodots of 50, 100nm dot diameter with an inter-dot spacing of 20, 70nm and heights 40, 100nm respectively, were engineered on Silicon substrates. MG63 cells were cultured for 72 hours and the modulation in morphology, focal adhesions, cell extensible area, cell viability, transcription factors and genes responsible for bone protein secretion as a function of the nanodot diameter, inter-dot distance and nanodot height were evaluated. Nanodots of 50nm diameter with a 20nm inter-dot spacing and 40nm height enhanced cell spreading area by 40%, promoted cell viability by 70% and upregulated transcription factors and genes twice as much, as compared to the 100nm nanodots with 70nm inter-dot spacing and 100nm height. Favorable interactions between cells and all dimensions of 50nm nanodot diameter were observed, determined with Scanning electron microscopy and Immunofluorescence staining. Nanodot height played a vital role in controlling the cell fate. Dimensions of nanodot features which triggered a transition in cell characteristics or behavior was also defined through statistical analysis. The findings of this study provide insights in the parameters of nanotopographic features which can vitally control the cell fate and should therefore be taken into account when designing implant geometries.
纳米拓扑线索可用于理解细胞与其微环境之间相互作用的本质，以生成更优的植入物几何形状。然而，调节细胞行为和特征（如粘着斑、细胞形态）的纳米表面参数尚未完全明确。在此，我们研究了不同纳米拓扑尺寸在调节细胞行为、特征以及最终细胞命运方面的作用，并据此提出了一种改善植入物表面几何形状的方法。分别在硅基板上制备了直径为50、100nm，点间距为20、70nm，高度为40、100nm的氧化钽纳米点。将MG63细胞培养72小时，并评估细胞形态、粘着斑、细胞可伸展面积、细胞活力、转录因子以及负责骨蛋白分泌的基因随纳米点直径、点间距和纳米点高度的变化情况。与直径100nm、点间距70nm、高度100nm的纳米点相比，直径50nm、点间距20nm、高度40nm的纳米点使细胞铺展面积增加了40%，细胞活力提高了70%，转录因子和基因上调了两倍。通过扫描电子显微镜和免疫荧光染色观察到细胞与直径50nm纳米点的所有尺寸之间存在良好的相互作用。纳米点高度在控制细胞命运方面起着至关重要的作用。通过统计分析还确定了引发细胞特征或行为转变的纳米点特征尺寸。本研究结果为纳米拓扑特征参数提供了见解，这些参数可至关重要地控制细胞命运，因此在设计植入物几何形状时应予以考虑。
