Fractal Dimension as a Measure of Altered Actin Cytoskeleton in MC3T3-E1 Cells Under Simulated Microgravity Using 3-D/2-D Clinostats
Fractal Dimension as a Measure of Altered Actin Cytoskeleton in MC3T3-E1 Cells Under Simulated Microgravity Using 3-D/2-D Clinostats
复制标题
使用 3-D/2-D 回转器在模拟微重力下用分形维数测量 MC3T3-E1 细胞中肌动蛋白细胞骨架的改变
DOI：
10.1109/tbme.2012.2187785
复制
发表时间：
2012-05-01
影响因子：
4.6
通讯作者：
Shang, Peng
中科院分区：
文献类型：
--
作者：
Qian, A. R.;Li, D.;Shang, Peng
成骨细胞，骨形成细胞，响应各种机械力，如拉伸和流体剪切力在本质上相似的方式。细胞骨架作为细胞的承重结构，对改变的惯性力很敏感。细胞骨架的破坏将导致细胞结构和功能的改变。然而，由于细胞骨架结构的复杂性，很难定量地说明细胞骨架重排。通常，由外部刺激引起的肌动蛋白组织的形态变化基本上是描述性的。在这项研究中，分形维数（D）分析用于量化的形态学变化的肌动蛋白细胞骨架成骨样细胞（MC3T3-E1）在模拟微重力下使用3-D/2-D回转器。采用ImageJ软件对肌动蛋白细胞骨架的分形维数进行盒计数法计算。实时荧光定量PCR和免疫荧光检测进一步证实了分形维数分析的结果。结果显示，与对照组相比，3D/2D旋转条件下培养24 h和48 h后，成骨样细胞肌动蛋白骨架的D值、β-actin mRNA的表达和F-actin的平均荧光强度均显著降低。研究结果表明，三维/二维旋转同时影响肌动蛋白细胞骨架的结构和mRNA的表达，分形可能是定量分析不同环境下细胞骨架变化的一种有前途的方法。
Osteoblasts, the bone-forming cells, respond to various mechanical forces, such as stretch and fluid shear force in essentially similar ways. The cytoskeleton, as the load-bearing architecture of the cell, is sensitive to altered inertial forces. Disruption of the cytoskeleton will result in alteration of cellular structure and function. However, it is difficult to quantitatively illustrate cytoskeletal rearrangement because of the complexity of cytoskeletal structure. Usually, the morphological changes in actin organization caused by external stimulus are basically descriptive. In this study, fractal dimensions (D) analysis was used to quantify the morphological changes in the actin cytoskeleton of osteoblast-like cells (MC3T3-E1) under simulated microgravity using 3-D/2-D clinostats. The ImageJ software was used to count the fractal dimension of actin cytoskeleton by box-counting methods. Real-time PCR and immunofluroscent assays were used to further confirm the results obtained by fractal dimension analysis. The results showed significant decreases in D value of actin cytoskeleton, β-actin mRNA expression, and the mean fluorescence intensity of F-actin in osteoblast-like cells after 24 or 48 h of incubation under 3-D/2-D clinorotation condition compared with control. The findings indicate that 3-D/2-D clinorotation affects both actin cytoskeleton architecture and mRNA expression, and fractal may be a promising approach for quantitative analysis of the changes in cytoskeleton in different environments.