Bone cell mechanosensation of fluid flow stimulation: a fluid–structure interaction model characterising the role integrin attachments and primary cilia

Load-induced fluid flow acts as an important biophysical signal for bone cell mechanotransduction in vivo, where the mechanical environment is thought to be monitored by integrin and primary cilia mechanoreceptors on the cell body. However, precisely how integrin- and primary cilia-based mechanosensors interact with the surrounding fluid flow stimulus and ultimately contribute to the biochemical response of bone cells within either the in vitro or in vivo environment remains poorly understood. In this study, we developed fluid–structure interaction models to characterise the deformation of integrin- and primary cilia-based mechanosensors in bone cells under fluid flow stimulation. Under in vitro fluid flow stimulation, these models predicted that integrin attachments on the cell–substrate interface were highly stimulated $$(\varepsilon _\mathrm{eq}> 200{,}000\,\upmu \upvarepsilon )$$(εeq>200,000με), while the presence of a primary cilium on the cell also resulted in significant strain amplifications, arising at the ciliary base. As such, these mechanosensors likely play a role in mediating bone mechanotransduction in vitro. Under in vivo fluid flow stimulation, integrin attachments along the canalicular wall were highly stimulated and likely play a role in mediating cellular responses in vivo. The role of the primary cilium as a flow sensor in vivo depended upon its configuration within the lacunar cavity. Specifically, our results showed that a short free-standing primary cilium could not effectively fulfil a flow sensing role in vivo. However, a primary cilium that discretely attaches the lacunar wall can be highly stimulated, due to hydrodynamic pressure in the lacunocanalicular system and, as such, could play a role in mediating bone mechanotransduction in vivo.

[1]  S. M. Sims,et al.  Estimating the sensitivity of mechanosensitive ion channels to membrane strain and tension. , 2004, Biophysical journal.

[2]  E H Burger,et al.  Differential stimulation of prostaglandin G/H synthase-2 in osteocytes and other osteogenic cells by pulsating fluid flow. , 2000, Biochemical and biophysical research communications.

[3]  A. Pitsillides,et al.  Mechanical strain‐induced NO production by bone cells: a possible role in adaptive bone (re)modeling? , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[4]  C. Jacobs,et al.  Primary Cilia-Mediated Mechanotransduction in Human Mesenchymal Stem Cells , 2012, Stem cells.

[5]  P. Nijweide,et al.  Pulsating fluid flow increases nitric oxide (NO) synthesis by osteocytes but not periosteal fibroblasts--correlation with prostaglandin upregulation. , 1995, Biochemical and biophysical research communications.

[6]  H J Donahue,et al.  Substrate deformation levels associated with routine physical activity are less stimulatory to bone cells relative to loading-induced oscillatory fluid flow. , 2000, Journal of biomechanical engineering.

[7]  H. Brismar,et al.  Mechanical properties of primary cilia regulate the response to fluid flow. , 2010, American journal of physiology. Renal physiology.

[8]  Amber L. Rath,et al.  Correlation of cell strain in single osteocytes with intracellular calcium, but not intracellular nitric oxide, in response to fluid flow. , 2010, Journal of biomechanics.

[9]  Theo H Smit,et al.  Nitric oxide production by bone cells is fluid shear stress rate dependent. , 2004, Biochemical and biophysical research communications.

[10]  S. Weinbaum,et al.  Attachment of Osteocyte Cell Processes to the Bone Matrix , 2009, Anatomical record.

[11]  A. van der Plas,et al.  Sensitivity of osteocytes to biomechanical stress in vitro , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[12]  Mia M. Thi,et al.  Mechanosensory responses of osteocytes to physiological forces occur along processes and not cell body and require αVβ3 integrin , 2013, Proceedings of the National Academy of Sciences.

[13]  Theo H Smit,et al.  Dynamic shear stress in parallel-plate flow chambers. , 2005, Journal of biomechanics.

[14]  T J Chambers,et al.  Induction of NO and prostaglandin E2 in osteoblasts by wall-shear stress but not mechanical strain. , 1997, American journal of physiology. Endocrinology and metabolism.

[15]  Ted J. Vaughan,et al.  Are all osteocytes equal? Multiscale modelling of cortical bone to characterise the mechanical stimulation of osteocytes , 2013, International journal for numerical methods in biomedical engineering.

[16]  Matthew E. Downs,et al.  An experimental and computational analysis of primary cilia deflection under fluid flow , 2014, Computer methods in biomechanics and biomedical engineering.

[17]  Kenneth R Spring,et al.  A physiological view of the primary cilium. , 2005, Annual review of physiology.

[18]  Stefaan W Verbruggen,et al.  Fluid flow in the osteocyte mechanical environment: a fluid–structure interaction approach , 2013, Biomechanics and Modeling in Mechanobiology.

[19]  C. Jacobs,et al.  β1 Integrins Mediate Mechanosensitive Signaling Pathways in Osteocytes , 2010, Calcified Tissue International.

[20]  C. Jacobs,et al.  Oscillating fluid flow regulates gap junction communication in osteocytic MLO-Y4 cells by an ERK1/2 MAP kinase-dependent mechanism. , 2003, Bone.

[21]  M. Schaffler,et al.  Osteocyte differentiation is regulated by extracellular matrix stiffness and intercellular separation. , 2013, Journal of the mechanical behavior of biomedical materials.

[22]  C. Jacobs,et al.  Deletion of β1 Integrins from Cortical Osteocytes Reduces Load-Induced Bone Formation , 2009 .

[23]  Masaki Hojo,et al.  Calcium response in single osteocytes to locally applied mechanical stimulus: differences in cell process and cell body. , 2009, Journal of biomechanics.

[24]  Matthew E. Downs,et al.  The mechanics of the primary cilium: an intricate structure with complex function. , 2012, Journal of biomechanics.

[25]  Anne M. Robertson,et al.  The numerical design of a parallel plate flow chamber for investigation of endothelial cell response to shear stress , 2003 .

[26]  Christopher R Jacobs,et al.  Primary cilium‐dependent mechanosensing is mediated by adenylyl cyclase 6 and cyclic AMP in bone cells , 2010, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  E. A. Schwartz,et al.  Analysis and modeling of the primary cilium bending response to fluid shear. , 1997, The American journal of physiology.

[28]  J. Shah,et al.  Identification of Signaling Pathways Regulating Primary Cilium Length and Flow-Mediated Adaptation , 2010, Current Biology.

[29]  T M Keaveny,et al.  Osteoblasts respond to pulsatile fluid flow with short-term increases in PGE(2) but no change in mineralization. , 2001, Journal of applied physiology.

[30]  D. Burr,et al.  Mechanotransduction in bone: osteoblasts are more responsive to fluid forces than mechanical strain. , 1997, The American journal of physiology.

[31]  Fumihiko Kajiya,et al.  The alteration of a mechanical property of bone cells during the process of changing from osteoblasts to osteocytes. , 2008, Bone.

[32]  C. Benhamou,et al.  Centrosome Fine Ultrastructure of the Osteocyte Mechanosensitive Primary Cilium , 2012, Microscopy and Microanalysis.

[33]  Minqi Li,et al.  Targeted ablation of osteocytes induces osteoporosis with defective mechanotransduction. , 2007, Cell metabolism.

[34]  Christopher S. Chen,et al.  Cell adhesion and mechanical stimulation in the regulation of mesenchymal stem cell differentiation , 2013, Journal of cellular and molecular medicine.

[35]  X. Guo,et al.  Osteocytic network is more responsive in calcium signaling than osteoblastic network under fluid flow , 2012, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[36]  Stefaan W Verbruggen,et al.  Strain amplification in bone mechanobiology: a computational investigation of the in vivo mechanics of osteocytes , 2012, Journal of The Royal Society Interface.

[37]  L. Lanyon,et al.  Cellular responses to mechanical loading in vitro , 1990, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[38]  C. Jacobs,et al.  A role for the primary cilium in paracrine signaling between mechanically stimulated osteocytes and mesenchymal stem cells. , 2011, Biochemical and biophysical research communications.

[39]  E. A. Tonna,et al.  Electron microscopy of aging skeletal cells. I. Centrioles and solitary cilia. , 1972, Journal of gerontology.

[40]  R. Kwon,et al.  Microfluidic Enhancement of Intramedullary Pressure Increases Interstitial Fluid Flow and Inhibits Bone Loss in Hindlimb Suspended Mice , 2010, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[41]  C. Jacobs,et al.  Mechanosensing by the Primary Cilium: Deletion of Kif3A Reduces Bone Formation Due to Loading , 2012, PloS one.

[42]  C. Hung,et al.  What is the role of the convective current density in the real-time calcium response of cultured bone cells to fluid flow? , 1996, Journal of biomechanics.

[43]  S. McGlashan,et al.  Localization of Extracellular Matrix Receptors on the Chondrocyte Primary Cilium , 2006, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[44]  R. Burgkart,et al.  Viscoelastic properties of the cell nucleus. , 2000, Biochemical and biophysical research communications.

[45]  Daniel A. Fletcher,et al.  A multi-structural single cell model of force-induced interactions of cytoskeletal components. , 2013, Biomaterials.

[46]  Daniel P Nicolella,et al.  Dark horse in osteocyte biology: Glycocalyx around the dendrites is critical for osteocyte mechanosensing. , 2011, Communicative & integrative biology.

[47]  R. Duncan,et al.  Parathyroid Hormone Enhances Fluid Shear‐Induced [Ca2+]i Signaling in Osteoblastic Cells Through Activation of Mechanosensitive and Voltage‐Sensitive Ca2+ Channels , 2001, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[48]  Min Jae Song,et al.  Mapping the Mechanome of Live Stem Cells Using a Novel Method to Measure Local Strain Fields In Situ at the Fluid-Cell Interface , 2012, PloS one.

[49]  Damien Lacroix,et al.  Structural finite element analysis to explain cell mechanics variability. , 2014, Journal of the mechanical behavior of biomedical materials.

[50]  Jenneke Klein-Nulend,et al.  A comparison of strain and fluid shear stress in stimulating bone cell responses—a computational and experimental study , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[51]  L. Bonewald,et al.  Expression of Functional Gap Junctions and Regulation by Fluid Flow in Osteocyte‐Like MLO‐Y4 Cells , 2001, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[52]  E H Burger,et al.  The production of nitric oxide and prostaglandin E(2) by primary bone cells is shear stress dependent. , 2001, Journal of biomechanics.

[53]  Ron Y Kwon,et al.  Primary cilia mediate mechanosensing in bone cells by a calcium-independent mechanism , 2007, Proceedings of the National Academy of Sciences.

[54]  T. Vaughan,et al.  Loading-Induced Interstitial Fluid Flow in Bone Mechanobiology: An FSI Approach to the Osteocyte Environment , 2012 .

[55]  Min Jae Song,et al.  In Situ Spatiotemporal Mapping of Flow Fields around Seeded Stem Cells at the Subcellular Length Scale , 2010, PloS one.

[56]  S. Chien,et al.  Integrin‐Mediated Expression of Bone Formation‐Related Genes in Osteoblast‐Like Cells in Response to Fluid Shear Stress: Roles of Extracellular Matrix, Shc, and Mitogen‐Activated Protein Kinase , 2008, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[57]  Eric J. Anderson,et al.  Idealization of pericellular fluid space geometry and dimension results in a profound underprediction of nano-microscale stresses imparted by fluid drag on osteocytes. , 2008, Journal of biomechanics.

[58]  S. Bowser,et al.  Ultrastructural, tomographic and confocal imaging of the chondrocyte primary cilium in situ , 2004, Cell biology international.

[59]  Jean X. Jiang,et al.  Dark horse in osteocyte biology , 2011 .

[60]  Joseph D. Gardinier,et al.  Cyclic Hydraulic Pressure and Fluid Flow Differentially Modulate Cytoskeleton Re-Organization in MC3T3 Osteoblasts , 2009, Cellular and molecular bioengineering.

[61]  S. Weinbaum,et al.  A model for the role of integrins in flow induced mechanotransduction in osteocytes , 2007, Proceedings of the National Academy of Sciences.

[62]  S. Weinbaum,et al.  On the electrophysiological response of bone cells using a Stokesian fluid stimulus probe for delivery of quantifiable localized picoNewton level forces. , 2011, Journal of biomechanics.

[63]  L. Lanyon Functional strain as a determinant for bone remodeling , 2006, Calcified Tissue International.