Osteoblast, playing a key role in the pathophysiology of osteoporosis, is one of the mechanical stress sensitive cells. The effects of mechanical load-induced changes of mechanical properties in osteoblast cells were studied at real-time. Osteoblasts obtained from young Wistar rats were exposed to mechanical loads in different frequencies and resting intervals generated by atomic force microscopy (AFM) probe tip and simultaneously measured the changes of the mechanical properties by AFM. The enhancement of the mechanical properties was observed and quantified by the increment of the apparent Young's modulus, E*. The observed mechanical property depended on the frequency of applied tapping loads. For the resting interval is 50s, the mechanical load-induced enhancement of E*-values disappears. It seems that the enhanced mechanical property was recover able under no additional mechanical stimulus.