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Updated: Jun 20, 2026
A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
Published on: March 15, 2018
Effects of high frequency loading on RANKL and OPG mRNA expression in ST-2 murine stromal cells
Chi Hyun Kim1, Kyung Hwan Kim, Christopher R Jacobs
1Department of Biomedical Engineering, Yonsei University, Gangwon Do, Korea. chihyun@yonsei.ac.kr
Background:
Oscillatory fluid flow (OFF)-induced shear stress leads to positive bone remodeling through pro-formative and anti-resorptive effects on bone cells. In this study, the effects of high frequency OFF on expression of receptor activator of NF-kappaB ligand (RANKL) and osteoprotegerin (OPG), two important regulators of osteoclast differentiation, were investigated.
Methods:
Cells were exposed to 1 Pa peak shear stress using three loading frequencies (1, 10, and 20 Hz) widely employed in cell, animal, and clinical studies of bone remodeling. Two separate experiments were performed where either the total number of cycles (3600 cycles) or the total loading time (60 min) was kept constant. Real-time RT-PCR was used to quantify mRNA levels of RANKL, OPG.
Results:
3600 cycles of OFF at 1 Hz and 10 Hz loading decreased RANKL/OPG ratio. Interestingly, these results were due to different mechanisms where at 1 Hz the decrease was due to an increase in OPG mRNA, whereas at 10 Hz the decrease was due to a decrease in RANKL mRNA.
Conclusion:
Although high frequency OFF does not appear to further enhance the decrease in the RANKL/OPG ratio, these results suggest a potential to differentially control the change in either RANKL or OPG mRNA expression by applying different loading frequencies.
