Spontaneous cellular vibratory motions of osteocytes are regulated by ATP and spectrin network

Vibration at high frequency has been demonstrated to be anabolic for bone and embedded osteocytes. The response of osteocytes to vibration is frequency-dependent, but the mechanism remains unclear. Our previous computational study using an osteocyte finite element model has predicted a resonance effect involving in the frequency-dependent response of osteocytes to vibration. However, the cellular spontaneous vibratory motion of osteocytes has not been confirmed. In the present study, the cellular vibratory motions (CVM) of osteocytes were recorded by a custom-built digital holographic microscopy and quantitatively analyzed. The roles of ATP and spectrin network in the CVM of osteocytes were studied. Results showed the MLO-Y4 osteocytes displayed dynamic vibratory motions with an amplitude of ~80 nm, which is relied both on the ATP content and spectrin network. Spectrum analysis showed several frequency peaks in CVM of MLO-Y4 osteocytes at 30 Hz, 39 Hz, 83 Hz and 89 Hz. These peak frequencies are close to the commonly used effective frequencies in animal training and in-vitro cell experiments, and show a correlation with the computational predictions of the osteocyte finite element model. These results implicate that osteocytes are dynamic and the cellular dynamic motion is involved in the cellular mechanotransduction of vibration.