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Previous studies demonstrated that extracellular calcium efflux ([Ca2+]E) originates from the regions of bone extracellular matrix that are undergoing microdamage. Such [Ca2+]E is reported to induce the activation of intracellular calcium signaling ([Ca2+]I) in MC3T3-E1 cells. The current study investigated the association between microdamage and local activation of intracellular calcium signaling quantifiably in MC3T3-E1 cells. Cells were seeded on devitalized notched bovine bone samples to induce damage controllably within the field of observation. A sequential staining procedure was implemented to stain for intracellular calcium activation followed by staining for microdamage on the same sample. The increase in [Ca2+]I fluorescence in cells of mechanically loaded samples was greater than that of unloaded negative control cells. The results showed that more than 80% of the cells with increased [Ca2+]I fluorescence were located within the damage zone. In conclusion, the findings demonstrate that there are spatial proximity between diffuse microdamage induction and the activation of intracellular calcium ([Ca2+]I) signaling in MC3T3-E1 cells. The downstream responses to the observed activation in future research may help understand how bone cells repair microdamage. Download references

Activation of intracellular calcium signaling in osteoblasts colocalizes with the formation of post-yield diffuse microdamage in bone matrix