Osteocyte calcium signaling response to bone matrix deformation
Osteocyte calcium signaling response to bone matrix deformation
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DOI：
10.1016/j.jbiomech.2009.07.006
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发表时间：
2009-11-13
影响因子：
2.4
通讯作者：
Kamioka, Hiroshi
中科院分区：
文献类型：
--
作者：
Adachi, Taiji;Aonuma, Yuki;Kamioka, Hiroshi
骨细胞被广泛认为在机械力传感中发挥重要作用，以实现在不断变化的力学环境中的适应性骨重建。体外研究已经阐明了几种类型的机械刺激，如静水压力，流体剪切应力和直接变形影响骨细胞的功能。然而，骨基质中骨细胞对机械刺激的反应还没有被清楚地了解。在本研究中，我们观察了骨细胞钙信号对定量施加的骨基质变形的反应。建立了一种新的实验系统，在显微镜载物台上对培养的骨组织和骨细胞施加变形。作为对骨基质中骨细胞的机械刺激，使用一对玻璃微针对从13天龄鸡胚颅骨获得的骨碎片施加面内剪切变形。骨基质和细胞的变形进行了定量评价，使用图像相关方法，通过应用差分干涉对比度图像的矩阵和免疫标记的骨细胞的荧光图像，连同成像的细胞钙瞬变使用比率法。结果证实，新开发的系统使我们能够在显微镜下成功地将变形应用于骨基质和骨细胞，而没有显著的焦平面移位或偏离观察视场。该系统可以为进一步开发的基础，通过检查各种类型的快速生化信号反应和基质变形诱导的细胞间通讯，研究骨基质中骨细胞的机械传感机制。(C)2009爱思唯尔有限公司保留所有权利。
Osteocytes embedded in calcified bone matrix have been widely believed to play important roles in mechanosensing to achieve adaptive bone remodeling in a changing mechanical environment. In vitro studies have clarified several types of mechanical stimuli such as hydrostatic pressure, fluid shear stress, and direct deformation influence osteocyte functions. However, osteocyte response to mechanical stimuli in the bone matrix has not been clearly understood. In this study, we observed the osteocyte calcium signaling response to the quantitatively applied deformation in the bone matrix. A novel experimental system was developed to apply deformation to cultured bone tissue with osteocytes on a microscope stage. As a mechanical stimulus to the osteocytes in bone matrix, in-plane shear deformation was applied using a pair of glass microneedles to bone fragments, obtained from 13-day-old embryonic chick calvariae. Deformation of bone matrix and cells was quantitatively evaluated using an image correlation method by applying for differential interference contrast images of the matrix and fluorescent images of immunolabeled osteocytes, together with imaging of the cellular calcium transient using a ratiometric method. As a result, it was confirmed that the newly developed system enables us to apply deformation to bone matrix and osteocytes successfully under the microscope without significant focal plane shift or deviation from the observation view field. The system could be a basis for further development to investigate the mechanosensing mechanism of osteocytes in bone matrix through examination of various types of rapid biochemical signaling responses and intercellular communication induced by matrix deformation. (C) 2009 Elsevier Ltd. All rights reserved.