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Hydraulic Pressure During Fluid Flow Regulates Purinergic Signaling and Cytoskeleton Organization of Osteoblasts

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Abstract

During physiological activities, osteoblasts experience a variety of mechanical forces that stimulate anabolic responses at the cellular level necessary for the formation of new bone. Previous studies have primarily investigated the osteoblastic response to individual forms of mechanical stimuli. However in this study, we evaluated the response of osteoblasts to two simultaneous, but independently controlled stimuli; fluid flow-induced shear stress (FSS) and static or cyclic hydrostatic pressure (SHP or CHP, respectively). MC3T3-E1 osteoblasts-like cells were subjected to 12 dynes/cm2 FSS along with SHP or CHP of varying magnitudes to determine if pressure enhances the anabolic response of osteoblasts during FSS. For both SHP and CHP, the magnitude of hydraulic pressure that induced the greatest release of ATP during FSS was 15 mmHg. Increasing the hydraulic pressure to 50 mmHg or 100 mmHg during FSS attenuated the ATP release compared to 15 mmHg during FSS. Decreasing the magnitude of pressure during FSS to atmospheric pressure reduced ATP release to that of basal ATP release from static cells and inhibited actin reorganization into stress fibers that normally occurred during FSS with 15 mmHg of pressure. In contrast, translocation of nuclear factor kappa B (NFκB) to the nucleus was independent of the magnitude of hydraulic pressure and was found to be mediated through the activation of phospholipase-C (PLC), but not src kinase. In conclusion, hydraulic pressure during FSS was found to regulate purinergic signaling and actin cytoskeleton reorganization in the osteoblasts in a biphasic manner, while FSS alone appeared to stimulate NFκB translocation. Understanding the effects of hydraulic pressure on the anabolic responses of osteoblasts during FSS may provide much needed insights into the physiologic effects of coupled mechanical stimuli on osteogenesis.

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Acknowledgments

This study was supported by funding from NIH/NIAMS (AR043222, AR051901, AR054385, P30GM103333, and AR064668).

Conflict of interest

The authors Joseph D. Gardinier, Vimal Gangadharan, Liyun Wang, and Randall L. Duncan declare that they have no conflicts of interest.

Ethical Standards

No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.

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Authors and Affiliations

  1. Biomechanics and Movement Science, University of Delaware, Newark, DE, 19716, USA

    Joseph D. Gardinier, Liyun Wang & Randall L. Duncan

  2. Department of Mechanical Engineering, University of Delaware, Newark, DE, 19716, USA

    Liyun Wang & Randall L. Duncan

  3. Department of Mechanical Biological Sciences, University of Delaware, Newark, DE, 19716, USA

    Vimal Gangadharan & Randall L. Duncan

  4. Department of Biological and Materials Science, University of Michigan, 2224 School of Dentistry, Ann Arbor, MI, 48109, USA

    Joseph D. Gardinier

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  1. Joseph D. Gardinier
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Correspondence to Joseph D. Gardinier.

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Associate Editor Michael R. King oversaw the review of this article.

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Gardinier, J.D., Gangadharan, V., Wang, L. et al. Hydraulic Pressure During Fluid Flow Regulates Purinergic Signaling and Cytoskeleton Organization of Osteoblasts. Cel. Mol. Bioeng. 7, 266–277 (2014). https://doi.org/10.1007/s12195-014-0329-8

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