Reduction of Bone Nodule Formation in MC3T3-E1 Cells by Treatment with Hydrogen Peroxide

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It is well known that bone formation steadily declines with age, resulting in a significant loss of bone mass. Cellular senescence-elevated oxidative stress has been shown to be related to the regulation of cellular functions in physiological and pathological conditions, and it has been clearly demonstrated that oxidative stress inhibits osteoblast differentiation and leads to a reduction in bone formation. However, the precise mechanism of the effect of reactive oxygen species (ROS) on the function of osteoblastic cells involved in bone nodule formation is not clear. In an attempt to establish a useful cell culture experimental model, we examined the effects of H2O2 on bone nodule formation in an established cell line, MC3T3-E1, following treatment with H2O2 using von Kossa staining. We found that cell viability was not affected by H2O2, though 400 μM suppressed the total number, as well total area and bone nodule area in MC3T3-E1 cells. Thus, a cell culture experimental model of bone formation during the aging process was successfully established using MC3T3-E1 cells with an H2O2 challenge without the damaging effects of proliferation or cell death. The present experimental model may be useful for studying the mechanism of decline of bone formation that accompanies aging.