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Article Abstract
Mechanical stimuli significantly influence bone remodeling, although the detailed molecular mechanisms involving changes in intracellular tension during osteoblast differentiation remain unclear. The present study was performed to investigate the role of intracellular tension in osteogenic differentiation by utilizing a newly established osteoblast-like cell line. In this study, we established a novel osteoblast-like cell line derived from calvarial explants of transgenic mice ubiquitously expressing a Förster Resonance Energy Transfer (FRET)-based tension sensor, capable of real-time measurement of intracellular tension and mineralization. The established cell line FRET1-MC8 displayed superior proliferative ability compared to conventional MC3T3-E1 osteoblast-like cells, maintaining stable growth and mineralization capability through at least passage 65. Osteogenic medium (OM) significantly enhanced the expression of osteogenic differentiation markers and , with clear mineralization observed as early as day 7 and extensive mineralization by day 14, comparable to MC3T3-E1 cells. Scratch experiments revealed increase in apparent contractility in OM-cultured cells, and subsequent quantitative analysis using acceptor photobleaching-based FRET efficiency confirmed significantly increased intracellular tension at day 3 of osteogenic induction. This elevation in intracellular tension coincided with increased expression, suggesting a critical role of tension in promoting osteogenic differentiation. The use of a FRET-based tension sensor enabled fast and direct monitoring of intracellular tension and is applicable to real-time analysis using live-cell FRET imaging. The established FRET1-MC8 cell line provides a powerful research tool for direct measurement of intracellular tension during osteogenic differentiation, thereby contributing to a better understanding of the mechanical regulation of bone remodeling.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC12270729 | PMC |
| http://dx.doi.org/10.1016/j.bbrep.2025.102131 | DOI Listing |
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