Automatic Actin Filament Quantification of Osteoblasts and Their Morphometric Analysis on Microtextured Silicon-Titanium Arrays
Automatic Actin Filament Quantification of Osteoblasts and Their Morphometric Analysis on Microtextured Silicon-Titanium Arrays
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DOI：
10.3390/ma5071176
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发表时间：
2012-07-01
期刊：
影响因子：
3.4
通讯作者：
Nebe, J. Barbara
中科院分区：
文献类型：
--
作者：
Matschegewski, Claudia;Staehlke, Susanne;Nebe, J. Barbara
植入物表面的微纹理化在奋进改进生物相关植入物设计中具有重要意义。为了阐明生物材料的形貌对细胞生理学的作用，获得细胞行为和不同的微结构特性之间的定量相关性是非常必要的。到目前为止，显微镜下观察到的结构化生物材料上细胞骨架的重组一直难以转化为数据。我们使用几何微织构硅钛阵列作为模型系统。通过对硅晶片进行深度反应离子蚀刻来制备样品，形成矩形凹槽（宽度和高度：2 μ m）和立方柱（柱尺寸：2 x 2 x 5和5 x 5 x 5 μ m）;最后溅射涂覆100纳米钛。我们专注于MG-63成骨细胞的形态计量学分析，包括肌动蛋白细胞骨架的定量。通过我们的新软件FilaQuant，特别是开发的自动肌动蛋白丝识别，我们首先能够量化的肌动蛋白网络的变化依赖于材料表面的微观纹理。细胞的肌动蛋白纤维的长度显着减少的柱状表面相比，凹槽阵列（4-5倍），并完全重组的微柱上，但没有改变细胞的方向。我们的形态功能方法为细胞-材料相互作用的数据相关性开辟了新的可能性。
Microtexturing of implant surfaces is of major relevance in the endeavor to improve biorelevant implant designs. In order to elucidate the role of biomaterial's topography on cell physiology, obtaining quantitative correlations between cellular behavior and distinct microarchitectural properties is in great demand. Until now, the microscopically observed reorganization of the cytoskeleton on structured biomaterials has been difficult to convert into data. We used geometrically microtextured silicon-titanium arrays as a model system. Samples were prepared by deep reactive-ion etching of silicon wafers, resulting in rectangular grooves (width and height: 2 mu m) and cubic pillars (pillar dimensions: 2 x 2 x 5 and 5 x 5 x 5 mu m); finally sputter-coated with 100 nm titanium. We focused on the morphometric analysis of MG-63 osteoblasts, including a quantification of the actin cytoskeleton. By means of our novel software FilaQuant, especially developed for automatic actin filament recognition, we were first able to quantify the alterations of the actin network dependent on the microtexture of a material surface. The cells' actin fibers were significantly reduced in length on the pillared surfaces versus the grooved array (4-5 fold) and completely reorganized on the micropillars, but without altering the orientation of cells. Our morpho-functional approach opens new possibilities for the data correlation of cell-material interactions.