细胞单层铺展形状对破骨前体细胞融合状态的调控作用及其机制

发布时间：2018-05-03 03:05

本文选题：微模式 + 粘附形状　；参考：《北京理工大学》2016年硕士论文

【摘要】：老龄化、妇女绝经、病人长期卧床以及宇航员长期在太空中失重等都会造成骨量的大量流失,从而严重影响人们的健康。骨吸收和骨形成是骨重建的两个重要过程,其中骨吸收由破骨细胞完成。破骨前体细胞来源于骨髓造血干细胞,其为单核巨噬细胞,已经发现可在化学因子的诱导下分化为多核的成熟破骨细胞,而细胞所处的力学微环境是否参与调控破骨细胞的融合则不清楚。我们此前已经发现,单层铺展的细胞在圆环和圆形模式的边界处其细胞单层内的剪应力最大,因此拟研究这种局部力学微环境是否影响破骨前体细胞的融合以及其机制是什么。本文利用微接触印刷技术构建了多细胞单层铺展的圆环形和圆形微模式,然后观测了内外不同区域的细胞融合率分布,进而利用免疫荧光染色技术观察了F-actin、Vinculin、β-catenin、E-cadherin四种蛋白在圆环形和圆形微模式上不同位置的表达。实验结果显示,用MC3T3-E1上清液诱导破骨前体细胞在圆环和圆形微模式上融合4天和6后,在圆环和圆形的最外侧融合率最低;而F-actin、Vinculin、β-catenin、E-cadherin四种蛋白在圆环形和圆形微模式最外侧表达也最高。因此我们推测,细胞单层的外边界处细胞会受到较高水平的剪应力,从而细胞骨架张力、细胞与基底间的作用力以及细胞间的相互作用力都较大,最终导致破骨细胞融合率较小。以上实验结果表明细胞单层内的力学微环境对破骨前体细胞的融合具有重要作用,并初步阐明了调控此融合过程的分子机制。
[Abstract]:Aging, menopause, prolonged bed rest and weightlessness of astronauts in space can cause massive bone loss, which can seriously affect people's health. Bone resorption and bone formation are two important processes of bone reconstruction, in which bone resorption is accomplished by osteoclasts. Osteoclast derived from bone marrow hematopoietic stem cells, which are mononuclear macrophages, have been found to differentiate into multinucleated mature osteoclasts induced by chemical factors. Whether the mechanical microenvironment in which the cells are located is involved in the regulation of osteoclast fusion is unclear. We have previously found that monolayer cells exhibit the greatest shear stress in the monolayer at the boundaries of the circular and circular patterns, so we are going to study whether this local mechanical microenvironment affects the fusion of osteoclast precursor cells and what the mechanism is. In this paper, the circular and circular micromodels of multi-cell monolayer were constructed by microcontact printing, and the distribution of cell fusion rate in different regions of the cell was observed. The expression of four proteins of F-actinine Vinculin, 尾 -cateninine E-cadherin in circular and circular micropatterns was observed by immunofluorescence staining. The results showed that the fusion rate of osteoclast precursor cells induced by MC3T3-E1 supernatant was the lowest in circle and circle after 4 days and 6 days. The expression of F-actinin Vinculin, 尾 -cateninine E-cadherin was also the highest in the outermost of circular and circular micropatterns. So we speculate that the outer boundary of monolayer cells will be subjected to a higher level of shear stress, thus the cytoskeleton tension, the force between the cell and the substrate, and the interaction between the cells are larger. Finally, the rate of osteoclast fusion is small. The above results indicate that the mechanical microenvironment in the monolayer plays an important role in the fusion of osteoclast precursor cells, and the molecular mechanism that regulates the fusion process is elucidated.
【学位授予单位】：北京理工大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R580

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