Figure 1. Overview of the methods in this study.

(A) The continuous compressive system and (B) the overview of the experiment design. In the present study, MLO-Y4 osteocyte-like cells were proliferated under a continuous compressive force (≈3.1 Pa or 31 dynes/cm²) throughout the entire experiment after seeding the cells 3 h. A long-duration (36 h) additional continuous compressive force (CCF) at 3.2 Pa was applied as the treatment of loading history, followed by re-applying the same additional CCF (≈3.2 Pa or 32 dynes/cm²) for a short duration (1 h), to assess the effect of loading history on the expression of interesting genes as well as the cell number, viability, morphology, and cell-to-cell communications of MLO-Y4 osteocyte-like cells.