Real-time calcium response of cultured bone cells to fluid flow

Hung, Clark, T.; Pollack, Solomon, R.; Reilly, Thomas M.; Brighton, Carl, T.

Affiliations

¹Department of Bioengineering, University of Pennsylvania, Philadelphia, PA

²Department of Orthopaedic Surgery, University of Pennsylvania, Philadelphia, PA

Abstract

Using a parallel-plate flow chamber and fura-2 fluorescence microscopy, intracellular calcium was measured cell by cell in preconfluent primary culture rat calvarial bone cells to 18, 35, and 70 dynes/cm² of fluid-induced shear stress. A heterogeneous response with respect to peak amplitude and latency was observed for the culture, with an overriding dose-dependent relationship between the mean peak amplitude of response and shearstress magnitude. A dose dependence was observed between the number of responsive cells (responding >50% over basal levels) and shear-stress magnitude. Not all cells could be restimulated by repeated exposure to flow. The observed cell response appears to be independent of whether cells are clustered together or isolated. Substratum stretch, hydrostatic pressure, and fluid shear stress have been shown in the literature to increase inositol phosphate (IP,3) in bone cells, with IP₃ causing the release of calcium from intracellular stores such as the endoplasmic reticulum. Therefore, a 6-fold inhibitory effect observed when calcium release from stores was blocked with 8-(n,N-diethylamino)octyl 1-3,4,5-atrimethoxybenzoate hydrochloride implicates an IP₃ biochemical pathway mediating the fluid flow response in bone cells.

Links

PubMed: 7641488

WoS: A1995QR80500035

History

Received: 1994-04-28

Revised: 1994-07-29

Accepted: 1994-11-14

Cited Works (17)

Year	Entry
1985	Levesque MJ, Nerem RM. The elongation and orientation of cultured endothelial cells in response to shear stress. J Biomech Eng. November 1985;107(4):341-347.
1892	Wolff J. Das Gesetz der Transformation der Knochen. Berlin: Hirschwald; 1892.
1994	Weinbaum S, Cowin SC, Zeng Y. A model for the excitation of osteocytes by mechanical loading-induced bone fluid shear stresses. J Biomech. March 1994;27(3):339-360.
1987	Salzstein RA, Pollack SR, Mak AFT, Petrov N. Electromechanical potentials in cortical bone, I: a continuum approach. J Biomech. 1987;20(3):261-270.
1984	Lanyon LE. Functional strain as a determinant for bone remodeling. Calcif Tiss Int. March 1984;36(suppl 1):S56-S61.
1991	Turner CH. Homeostatic control of bone structure: an application of feedback theory. Bone. 1991;12(3):203-217.
1990	Murray DW, Rushton N. The effect of strain on bone cell prostaglandin E₂ release: a new experimental method. Calcif Tiss Int. July 1990;47(1):35-39.
1982	Gross D, Williams WS. Streaming potential and the electromechanical response of physiologically-moist bone. J Biomech. 1982;15(4):277-295.
1984	Yeh C-K, Rodan GA. Tensile forces enhance prostaglandin E synthesis in osteoblastic cells grown on collagen ribbons. Calcif Tiss Int. March 1984;36(1):S67-S71.
1990	Rubin CT, McLeod KJ, Bain SD. Functional strains and cortical bone adaptation: epigenetic assurance of skeletal integrity. J Biomech. 1990;23(suppl 1):43-54.
1994	Williams JL, Iannotti JP, Ham A, Bleuit J, Chen JH. Effects of fluid shear stress on bone cells. Biorheology. March–April 1994;31(2):163-170.
1990	Reich KM, Gay CV, Frangos JA. Fluid shear stress as a mediator of osteoblast cyclic adenosine monophosphate production. J Cell Physiol. April 1990;143(1):100-103.
1993	Reich KM, Frangos JA. Protein kinase C mediates flow-induced prostaglandin E₂ production in osteoblasts. Calcif Tiss Int. January 1993;52(1):62-66.
1991	Cowin SC, Moss-Salentijn L, Moss ML. Candidates for the mechanosensory system in bone. J Biomech Eng. May 1991;113(2):191-197.
1982	O'Connor JA, Lanyon LE, MacFie H. The influence of strain rate on adaptive bone remodelling. J Biomech. 1982;15(10):767-781.
1983	Frost HM. A determinant of bone architecture: the minimum effective strain. Clin Orthop Relat Res. August 1983;175:286-292.
1984	Pollack SR, Petrov N, Salzstein R, Brankov G, Blagoeva R. An anatomical model for streaming potentials in osteons. J Biomech. 1984;17(8):627-636.

Cited By (65)

Year	Entry
2000	Chen NX, Ryder KD, Pavalko FM, Turner CH, Burr DB, Qiu J, Duncan RL. Ca²⁺ regulates fluid shear-induced cytoskeletal reorganization and gene expression in osteoblasts. Am J Physiol Cell Physiol. May 2000;278(5):C989-C997.
2001	Donahue SW, Jacobs CR, Donahue HJ. Flow-induced calcium oscillations in rat osteoblasts are age, loading frequency, and shear stress dependent. Am J Physiol Cell Physiol. November 2001;281(5):C1635-C1641.
1999	Ajubi NE, Klein-Nulend J, Alblas MJ, Burger EH, Nijweide PJ. Signal transduction pathways involved in fluid flow-induced PGE₂ production by cultured osteocytes. Am J Physiol Endocrinol Metab. January 1999;276(1):E171-E178.
2008	Gurkan UA, Akkus O. The mechanical environment of bone marrow: a review. Ann Biomed Eng. December 2008;36(12):1978-1991.
2009	Fritton SP, Weinbaum S. Fluid and solute transport in bone: flow-induced mechanotransduction. Annu Rev Fluid Mech. 2009;41:347-374.
2000	Donahue HJ. Gap junctions and biophysical regulation of bone cell differentiation. Bone. May 2000;26(5):417-422.
2013	Klein-Nulend J, Bakker AD, Bacabac RG, Vatsa A, Weinbaum S. Mechanosensation and transduction in osteocytes. Bone. June 2013;54(2):182-190.
2021	Harris TL, Silva MJ. Gene expression of intracortical bone demonstrates loading-induced increases in Wnt1 and Ngf and inhibition of bone remodeling processes. Bone. September 2021;150:116019.
1999	Burger EH, Klein-Nulend J. Mechanotransduction in bone: role of the lacunocanalicular network. FASEB J. May 1999;12(9001):S101-S112.
2006	Rubin J, Rubin C, Jacobs CR. Molecular pathways mediating mechanical signaling in bone. Gene. February 15, 2006;367:1-16.
2012	Thompson WR, Rubin CT, Rubin J. Mechanical regulation of signaling pathways in bone. Gene. July 25, 2012;503(2):179-193.
2001	Roberts SR, Knight MM, Lee DA, Bader DL. Mechanical compression influences intracellular Ca²⁺ signaling in chondrocytes seeded in agarose constructs. J Appl Physiol. April 2001;90(4):1385-1391.
2001	You J, Reilly GC, Zhen X, Yellowley CE, Chen Q, Donahue HJ, Jacobs CR. Osteopontin gene regulation by oscillatory fluid flow via intracellular calcium mobilization and activation of mitogen-activated protein kinase in MC3T3–E1 osteoblasts. J Biol Chem. April 20, 2001;276(16):13365-13371.
1996	Hung CT, Allen FD, Pollack SR, Brighton CT. What is the role of the convective current density in the real-time calcium response of cultured bone cells to fluid flow? J Biomech. November 1996;29(11):1403-1409.
1996	Hung CT, Allen FD, Pollack SR, Brighton CT. Intracellular Ca²⁺ stores and extracellular Ca²⁺ are required in the real-time Ca²⁺ response of bone cells experiencing fluid flow. J Biomech. November 1996;29(11):1411-1417.
1998	Jacobs CR, Yellowley CE, Davis BR, Zhou Z, Cimbala JM, Donahue HJ. Differential effect of steady versus oscillating flow on bone cells. J Biomech. November 1998;31(11):969-976.
1999	Wang L, Fritton SP, Cowin SC, Weinbaum S. Fluid pressure relaxation depends upon osteonal microstructure: modeling an oscillatory bending experiment. J Biomech. July 1999;32(7):663-672.
2000	Brown TD. Techniques for mechanical stimulation of cells in vitro: a review. J Biomech. January 2000;33(1):3-14.
2000	Hung CT, Ross Henshaw D, Wang CC-B, Mauck RL, Raia F, Palmer G, Grace Chao P-H, Mow VC, Ratcliffe A, Valhmu WB. Mitogen-activated protein kinase signaling in bovine articular chondrocytes in response to fluid flow does not require calcium mobilization. J Biomech. January 2000;33(1):73-80.
2000	Fritton SP, McLeod KJ, Rubin CT. Quantifying the strain history of bone: spatial uniformity and self-similarity of low-magnitude strains. J Biomech. March 2000;33(3):317-325.
2003	Donahue SW, Donahue HJ, Jacobs CR. Osteoblastic cells have refractory periods for fluid-flow-induced intracellular calcium oscillations for short bouts of flow and display multiple low-magnitude oscillations during long-term flow. J Biomech. January 2003;36(1):35-43.
2003	Haut Donahue TL, Haut TR, Yellowley CE, Donahue HJ, Jacobs CR. Mechanosensitivity of bone cells to oscillating fluid flow induced shear stress may be modulated by chemotransport. J Biomech. September 2003;36(9):1363-1371.
2009	Adachi T, Aonuma Y, Tanaka M, Hojo M, Takano-Yamamoto T, Kamioka H. Calcium response in single osteocytes to locally applied mechanical stimulus: differences in cell process and cell body. J Biomech. August 25, 2009;42(12):1989-1995.
2000	You J, Yellowley CE, Donahue HJ, Zhang Y, Chen Q, Jacobs CR. Substrate deformation levels associated with routine physical activity are less stimulatory to bone cells relative to loading-induced oscillatory fluid flow. J Biomech Eng. August 2000;122(4):387-393.
2015	Oftadeh R, Perez-Viloria M, Villa-Camacho JC, Vaziri A, Nazarian A. Biomechanics and mechanobiology of trabecular bone: a review. J Biomech Eng. January 2015;137(1):010802.
1999	McAllister TN, Frangos JA. Steady and transient fluid shear stress stimulate NO release in osteoblasts through distinct biochemical pathways. J Bone Miner Res. June 1999;14(6):930-936.
2001	Ryder KD, Duncan RL. Parathyroid hormone enhances fluid shear-induced [Ca²⁺]_i signaling in osteoblastic cells through activation of mechanosensitive and voltage-sensitive Ca²⁺ channels. J Bone Miner Res. February 2001;16(2):240-248.
2001	Hsieh Y, Turner CH. Effects of loading frequency on mechanically induced bone formation. J Bone Miner Res. May 2001;16(5):918-924.
2001	Lanyon L, Skerry T. Postmenopausal osteoporosis as a failure of bone's adaptation to functional loading: a hypothesis. J Bone Miner Res. November 2001;16(11):1937-1947.
2012	Lu XL, Huo B, Chiang V, Guo XE. Osteocytic network is more responsive in calcium signaling than osteoblastic network under fluid flow. J Bone Miner Res. March 2012;27(3):563-574.
2009	Riddle RC, Donahue HJ. From streaming-potentials to shear stress: 25 years of bone cell mechanotransduction. J Orthop Res. February 2009;27(2):143-149.
2002	Ehrlich PJ, Lanyon LE. Mechanical strain and bone cell function: a review. Osteoporos Int. September 2002;13(9):688-700.
2006	Gemmiti CV, Guldberg RE. Fluid flow increases type II collagen deposition and tensile mechanical properties in bioreactor-grown tissue-engineered cartilage. Tissue Eng. March 2006;12(3):469-479.
2010	Jähn K. Ex Vivo and in Vitro Culture Models of Human Osteoblast-Like Cells and the Effects of TGFβ₃ and Serum-Free Culture [PhD thesis]. Cardiff University; 2010.
2003	Kim CH. In Vivo Trabecular Bone Response to Mechanical Loading and Parathyroid Hormone Stimulation [PhD thesis]. Columbia University; 2003.
2005	Chao P-hG. Physical Forces on Cells: Effects of Applied Osmotic Loading and Direct Current Electric Fields [PhD thesis]. Columbia University; 2005.
2005	Takai E. Modulation of Bone Cell Mechanotransduction [PhD thesis]. Columbia University; 2005.
2009	Chan M. Mechanobiology of 3D Co-Culture Trabecular Explant Model [PhD thesis]. Columbia University; 2009.
2014	Lee KL. A Mechanism of Mechanotransduction Mediated by the Primary Cilium [PhD thesis]. Columbia University; 2014.
2016	Brown GN. The Sustainment and Consequences of Cytosolic Calcium Signals in Osteocytes [PhD thesis]. Columbia University; 2016.
2019	Campi AE. Mechanosensitive Ca²⁺ Signaling of Ex Vivo Osteocytes in Aging and Treatment [PhD thesis]. Columbia University; 2019.
2019	Estell EG. Modulation of Synovium Mechanobiology and Tribology in the Osteoarthritic Environment [PhD thesis]. Columbia University; 2019.
2010	Degala S. Fluid Flow Regulates Articular Chondrocyte Mechanotransduction Pathways in 3D Culture [PhD thesis]. Ithaca, NY: Cornell University; May 2010.
2002	Wang L. Investigating the Role of Interstitial Fluid Flow in Bone Adaptation and Metabolism [PhD thesis]. New York, NY: The City University of New York; 2002.
2002	You L. A New View of Mechanotransduction in Bone Cells [PhD thesis]. New York, NY: The City University of New York; 2002.
2004	Mi LY. Analysis of Avian Bone Response to Mechanical Loading Using a Computational Connected Network [PhD thesis]. New York, NY: The City College of New York; 2004.
2004	Thi MM. Cellular Communication and Mechanotransduction in Response to Fluid Shear Stress [PhD thesis]. New York, NY: The City University of New York; 2004.
2006	Han Y. On the Transmission of Fluid Forces to the Actin Cytoskeleton of Bone and Endothelial Cells [PhD thesis]. New York, NY: The City University of New York; 2006.
2006	Gemmiti CV. Effect of Fluid Flow on Tissue-Engineered Cartilage in a Novel Bioreactor [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; December 2006.
2000	Ryder KD. Parathyroid Hormone Modulates the Response of Osteoblast-Like Cells to Mechanical Stimulation [PhD thesis]. Indianapolis, IN: Indiana University; May 2000.
2018	Mikolajewicz N. Purinergic Mechanotransduction in Bone [PhD thesis]. McGill University; December 2018.
2016	Druchok C. Temporal Influence of NSAIDs on Mechanically Induced Bone Formation and Fluid Flow Stimulated Cellular PGE₂ Production [PhD thesis]. Hamilton, ON: McMaster University; August 2016.
2007	Lavagnino M. The Mechanotransduction Response of Tendon Cells to Tensile Loading [PhD thesis]. East Lansing, MI: Michigan State University; 2007.
2015	Oftadeh R. Hierarchical Analysis and Multiscale Modelling of Cellular Structures: From Meta Materials to Bone Structure [PhD thesis]. Northeastern University; December 2015.
2010	Gurkan UA. Engineering of Bone Marrow in Vitro for Investigating the Role of Growth Factors and Their Mechanoresponsiveness in Osteogenesis [PhD thesis]. Purdue University; May 2010.
2018	Canelón SP. Characterization of Type I Collagen and Osteoblast Response to Mechanical Loading [PhD thesis]. Purdue University; May 2018.
2006	Hannay G. Mechanical and Electrical Environments to Stimulate Bone Cell Development [PhD thesis]. Queensland University of Technology; August 2006.
2016	Wittkowske C. The Role of Mechanical Forces in Bone Formation: Evaluation of Long-Term Responses by Osteoblasts to Low Fluid Shear Stress in Vitro [PhD thesis]. University of Sheffield; September 2016.
2008	Kwon RY. Mechanical Behavior and Early Molecular Signaling During Mechanotransduction of Fluid Flow in Bone Cells [PhD thesis]. Stanford University; August 2008.
2009	Litzenberger JB. The Role of Integrin Signaling in Bone Mechanotransduction [PhD thesis]. Stanford University; 2009.
2008	Mulvihill BMC. Computational Investigation Into the Mechanoregulation of Osteoporosis [PhD thesis]. Trinity College Dublin; December 2008.
2010	Thompson WR. Perlecan Modulates the Function of the Osteocyte Lacuno-Canalicular System [PhD thesis]. University of Delaware; 2010.
2003	Ominsky MS. Effects of Hydrostatic Pressure, Biaxial Strain, and Fluid Shear on Osteoblastic Cells: Mechanotransduction Via NF-ΚB, MAP Kinase, and AP-1 Pathways [PhD thesis]. University of Michigan; 2003.
2004	Taboas JM. Mechanobiologic Regulation of Skeletal Progenitor Cell Differentiation [PhD thesis]. University of Michigan; 2004.
2021	Harris TL. Mechanisms for Osteoblast and Osteocyte Initiation and Sustainment of Bone Formation in Young-Adult and Aged Mice [PhD thesis]. St. Louis, MO: Washington University in St. Louis; August 2021.