OCY454 Osteocytes as an in Vitro Cell Model for Bone Remodeling Under Mechanical Loading

ABSTRACT Osteocytes’ mechano‐regulation of bone formation and resorption is key to maintaining appropriate bone health. Although extensive in vitro studies have explored osteocyte mechanobiology using the well‐established MLO‐Y4 cell model, the low amount of sclerostin secreted by this cell line ren...

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Bibliographic Details
Published in:Journal of orthopaedic research 2019-08, Vol.37 (8), p.1681-1689
Main Authors: Xu, Liangcheng Henry, Shao, Han, Ma, Yu‐Heng V., You, Lidan
Format: Article
Language:English
Subjects:
Adaptor Proteins, Signal Transducing - metabolism
Adenosine Triphosphate - metabolism
Animals
Bone Remodeling
Calcium Signaling
Cell Differentiation
Cell Line
mechanotransduction
Mechanotransduction, Cellular
Mice
OCY454
osteoblast
osteocyte
Osteocytes - physiology
Weight-Bearing
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Summary:ABSTRACT Osteocytes’ mechano‐regulation of bone formation and resorption is key to maintaining appropriate bone health. Although extensive in vitro studies have explored osteocyte mechanobiology using the well‐established MLO‐Y4 cell model, the low amount of sclerostin secreted by this cell line renders it inadequate for studying cross‐talk between osteocytes and osteoblasts under mechanical loading. Here, we investigated the potential of the sclerostin‐expressing OCY454 osteocyte cell model in fulfilling this role. Fully differentiated OCY454 cells were tested for mechano‐sensitivity by measuring changes in protein secretion, total adenosine triphosphate (ATP) content, and intracellular calcium in response to oscillatory fluid flow. Increases in sclerostin expression and total ATP content were observed. However, very low levels of receptor activator of the nuclear factor κ‐B ligand were detected, and there was a great inconsistency in calcium response. Conditioned medium (CM) from OCY454 cells was then used to culture osteoblast and osteoclast precursors. Osteoblast activity was quantified with alkaline phosphatase (ALP) and Alizarin Red S stain, while osteoclast differentiation was quantified with tartrate‐resistant acid phosphatase (TRAP) staining. We demonstrated that mechanically stimulated OCY454 cells released soluble factors that increased osteoblasts’ ALP activity (p 
ISSN:0736-0266
1554-527X
DOI:10.1002/jor.24302