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J. Biol. Chem., Vol. 276, Issue 16, 13365-13371, April 20, 2001
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From the Recently fluid flow has been shown to be a potent
physical stimulus in the regulation of bone cell metabolism. However,
most investigators have applied steady or pulsing flow profiles rather than oscillatory fluid flow, which occurs in vivo because
of mechanical loading. Here oscillatory fluid flow was demonstrated to
be a potentially important physical signal for loading-induced changes in bone cell metabolism. We selected three well known biological response variables including intracellular calcium
(Ca2+i), mitogen-activated protein
kinase (MAPK) activity, and osteopontin (OPN) mRNA levels to
examine the response of MC3T3-E1 osteoblastic cells to oscillatory
fluid flow with shear stresses ranging from 2 to
Osteopontin Gene Regulation by Oscillatory Fluid Flow via
Intracellular Calcium Mobilization and Activation of
Mitogen-activated Protein Kinase in MC3T3-E1 Osteoblasts*
,,,,, and
**
Musculoskeletal Research Laboratory,
Department of Orthopaedics and Rehabilitation, The Pennsylvania State
University College of Medicine, Hershey, Pennsylvania 17033, § Department of Pharmacology and Physiology, MCP-Hahneman
School of Medicine, Drexel University, Philadelphia, Pennsylvania
19129, ¶ Biomechanical Engineering Division, Department of
Mechanical Engineering, Stanford University, Stanford, California
94305, and Rehabilitation Research and Development
Center, Palo Alto Health Care System, Department of Veterans Affairs,
Palo Alto, California 94304
2
Newtons/m2 at 1 Hz, which is in the range expected
to occur during routine physical activities. Our results showed that
within 1 min, oscillatory flow induced cell
Ca2+i mobilization, whereas two
MAPKs (ERK and p38) were activated over a 2-h time frame. However,
there was no activation of JNK. Furthermore 2 h of oscillatory
fluid flow increased steady-state OPN mRNA expression levels by
approximately 4-fold, 24 h after exposure to fluid flow. The
presence of both ERK and p38 inhibitors and thapsigargin completely
abolished the effect of oscillatory flow on steady-state OPN mRNA
levels. In addition, experiments using a variety of pharmacological
agents suggest that oscillatory flow induces
Ca2+i mobilization via the L-type
voltage-operated calcium channel and the inositol
1,4,5-trisphosphate pathway.
*
This work was supported by National Institutes of Health
Grants AR45989, AG13087, AG00811, and AG17021, by the Whitaker
Foundation, Arthritis Foundation, and the United States Army Medical
Research and Materiel Command Award DAMD 17-98-1-8509.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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