In order to establish the principle to create smart composite materials, it's necessary to study from living bone tissue that has composite-like structure and high-grade environmental adaptive function. In the mechanosensory mechanism of bone remodeling, the osteocyte network system embedded in bone matrix is considered as one of the important elements. This study observed the calcium response of osteocytes isolated from and embedded in living bone tissue matrix to mechanical stimulation. As a result, sensitivity of the cell process was relatively higher than that of the cell body. In addition, the role of cytoskeletal actin structure in the mechanosensory mechanism was discussed in terms of its localized distribution. Finally, we observed the calcium response of osteocytes embedded in living bone tissue matrix in situ. This knowledge can be applied to clarify the relation between macroscopic mechanical stimulation to bone and microscopic mechanosensing of osteocytes.