Mechanical stimulation plays an important role in the normal metabolism and reorganization of bone. Many kinds of stress sensitive cells, including osteoblasts, can perceive and transduce the mechanical signals. As the fibrous framework throughout a cell, cytoskeleton as well as extracellular matrix and integrin is one of the critical components in mechanotransduction. Mechanical stimulation can induce the rearrangement of the cytoskeleton, and the signals are transduced and transformed by the second messagers, finally resulting in the changes of gene expression.The family of Rho, protein kinase C and focal adhesion kinase are the main messages involved in this process. This paper summarizes the recent advances about the effects and possible mechanism of mechanical stimulation on osteoblast cytoskeleton.