Osteocyte network system embedded in bone matrix is considered as one of the important elements in the mechanosensory system in bone remodeling. In order to understand the mechanosensory mechanism of osteocytes network, it is essential to evaluate mechanical stimulation to osteocyte network quantitatively as well as to observe biochemical response of osteocyte network to deformation of bone matrix. This study aimed to develop the method to evaluate osteocyte network deformation against bone matrix deformation. Osteocyte network in chick calvarial bone tissue was visualized by immunostaining of cell membrane using OB7.3. Osteocyte network displacement induced by localized deformation to bone matrix using a glass microneedle was observed. Analysis of osteocyte network displacement was fairly successful using the image-correlation method. Applicability of the method newly developed in this study was discussed in combination with biochemical response observation of change in intracellular calcium ion concentration using the fluorescent ratiometry method, in order to investigate the relation between macroscopic deformation of bone matrix and microscopic cellular response.
