Substrate Deformation Levels Associated With Routine Physical Activity Are Less Stimulatory to Bone Cells Relative to Loading-Induced Oscillatory Fluid Flow

[+] Author and Article Information
J. You, C. E. Yellowley, H. J. Donahue, Y. Zhang, Q. Chen, C. R. Jacobs

Musculoskeletal Research Laboratory, Department of Orthopaedics and Rehabilitation, The Pennsylvania State University College of Medicine, Hershey, PA 17033

J Biomech Eng 122(4), 387-393 (Mar 22, 2000) (7 pages) doi:10.1115/1.1287161 History: Received July 21, 1999; Revised March 22, 2000
Copyright © 2000 by ASME
Your Session has timed out. Please sign back in to continue.


Morey,  E. R., and Baylink,  D. J., 1978, “Inhibition of Bone Formation During Spaceflight,” Science, 201, pp. 1138–1141.
Sessions,  N. D., Halloran,  B. P., Bikle,  D. D., Wronski,  T. J., Cone,  C. M., and Morey-Holton,  E. R., 1989, “Bone Response to Normal Weight Bearing After a Period of Skeletal Unloading,” Am. J. Physiol., 257 (also: Endocinol. Metab., 20 ), pp. E606–610.
McLeod, K. J., Donahue, H. J., Levin, P. E., and Rubin, C. T., 1991, “LowFrequency Sinusoidal Electric Fields Alter Calcium Fluctuations in Osteoblast-Like Cells,” in: Electromagnetics in Biology and Medicine, Brighton, C. T., and Pollack, S. R., eds., San Francisco, San Francisco, pp. 111–115.
Buckley,  M. J., Banes,  A. J., Levin,  L. G., Sumpio,  B. E., Sato,  M., Jordan,  R., Gilbert,  J., Link,  G. W., and Tran Son Tay,  R., 1988, “Osteoblasts Increase Their Rate of Division and Align in Response to Cyclic, Mechanical Tension in Vitro,” J. Bone Miner. Res., 4, pp. 225–236.
Rodan,  G. A., Bourret,  L. A., Harvey,  A., and Mensi,  T., 1975, “Cyclic AMP and Cyclic GMP: Mediators of the Mechanical Effects on Bone Remodeling,” Science, 189, No. 4201, pp. 467–469.
Toma,  C. D., Ashkar,  S., Gray,  M. L., Schaffer,  J. L., and Gerstenfeld,  L. C., 1997, “Signal Transduction of Mechanical Stimuli Is Dependent on Microfilament Integrity: Identification of Osteopontin as a Mechanically Induced Gene in Osteoblasts,” J. Bone Miner. Res., 12, pp. 1626–1636.
Burr,  D. B., Milgrom,  C., Fyhrie,  D., Forwood,  M. R., Nyska,  M., Finestone,  A., Hoshaw,  S., Saiag,  E., and Simkin,  A., 1996, “In Vivo Measurement of Human Tibial Strains During Vigorous Activity,” Bone, 18, pp. 405–410.
Stanford,  C. M., Stevens,  J. W., and Brand,  R. A., 1995, “Cellular Deformation Reversibly Depresses RT-PCR Detectable Levels of Bone-Related mRNA,” J. Biomech., 28, pp. 1419–1421.
Zaman,  G., Suswillo,  R. F. L., Cheng,  M. Z., Tavares,  I. A., and Lanyon,  L. E., 1997, “Early Responses to Dynamic Strain Change and Prostaglandins in Bone-Derived Cells in Culture,” J. Bone Miner. Res., 12, pp. 769–777.
Owan,  I., Burr,  D. B., Turner,  C. H., Qiu,  J., Tu,  Y., Onyia,  J. E., and Duncan,  R. L., 1997, “Mechanotransduction in Bone: Osteoblasts Are More Responsive to Fluid Forces Than Mechanical Strain,” Am. J. Physiol., 273, pp. C810–C815.
Cheng,  M. Z., Zaman,  G., Rawlinson,  S. C. F., Mohan,  S., Baylink,  D. J., and Lanyon,  L. E., 1999, “Mechanical Strain Stimulates ROS Cell Proliferation Through IGF-II and Estrogen Through IGF-I,” J. Bone Miner. Res., 14, pp. 1742–1750.
Kufahl,  R. H., and Saha,  S., 1990, “A Theoretical Model for Stress-Generated Fluid Flow in the Canaliculi–Lacunae Network in Bone Tissue,” J. Biomech., 23, No. 2, pp. 171–180.
Cowin,  S. C., Weinbaum,  S., and Zeng,  Y., 1995, “A Case for Bone Canaliculi as the Anatomical Site of Strain Generated Potentials,” J. Biomech., 28, pp. 1281–1297.
Weinbaum,  S., Cowin,  S. C., and Zeng,  Y. A., 1994, “A Model for Excitation of Osteocytes by Mechanical Loading Induced Bone Fluid Shear Stress,” J. Biomech., 27, pp. 339–360.
Smalt,  R., Mitchell,  F. T., Howard,  R. L., and Chambers,  T. J., 1997, “Induction of NO and Prostaglandin E2 in Osteoblasts by Wall-Shear Stress but Not Mechanical Strain,” Am. J. Physiol., 273, No. 4, Pt. 1, pp. E751–E758.
Zhang,  D., Weinbaum,  S., and Cowin,  S. C., 1998, “Estimates of the Peak Pressure in Bone Pore Water,” ASME J. Biomech. Eng., 120, pp. 697–703.
Jacobs,  C. R., Yellowley,  C. E., Davis,  B. R., Zhou,  Z., Cimbala,  J. M., and Donahue,  H. J., 1998, “Differential Effect of Steady Versus Oscillating Flow on Bone Cells,” J. Biomech., 31, pp. 969–976.
Hung,  C. T., Pollack,  S. R., Reilly,  T. M., and Brighton,  C. T., 1995, “RealTime Calcium Response of Cultured Bone Cells to Fluid Flow,” Clin. Orthop., 313, pp. 256–269.
Denhardt,  D. T., and Guo,  X., 1993, “Osteopontin: a Protein With Diverse Functions,” FASEB J., 17, pp. 1476–1481.
Gerstenfeld,  L. C., Uporov,  T., Ashka,  S., Salih,  E., Gerstenfeld,  L. C., and Glimcher,  M. J., 1995, “Regulation of Avian Osteopontin Pre- and Posttranslational Expression in Skeletal Tissue,” Ann. NY Acad. Sci., 270, pp. 67–82.
Terai,  K., Takano-Yamamoto,  T., Ohba,  Y., Hiura,  K., Sugimoto,  M., Sato,  M., Kawahata,  H., Inaguma,  N., Kitamur,  Y., and Nomur,  S., 1999, “Role of Osteopontin in Bone Remodeling Caused by Mechanical Stress,” J. Bone Miner. Res., 14, pp. 839–849.
Harter,  L. V., Hruska,  K. A., and Duncan,  R. L., 1995, “Human Osteoblasts Like Cells Respond to Mechanical Strain With Increased Bone Matrix Protein Production Independent of Hormonal Regulation, ” Endocrinology, 136, pp. 528–535.
Kubota,  T. M., Yamauchi,  M., Onozaki,  S. S., Suzuki,  Y., and Sodek,  J., 1993, “Influence of an Intermittent Compressive Force on Matrix Expression by ROS 17/2.8 Cells With Selective Stimulation of Osteopontin,” Arch. Oral Biol., 38, pp. 23–30.
Harris,  S. A., Enger,  R. J., Riggs,  B. L., and Spelsberg,  T. C., 1995, “Development and Characterization of a Conditionally Immortalized Human Fetal Osteoblastic Cell Line,” J. Bone Miner. Res., 10, No. 2, pp. 178–186.
Donahue,  H. J., Zhou,  Z., Li,  Z., and McCauley,  L. K., 1997, “Age-Related Decreases in Stimulatory G Protein-Coupled Adenylate Cyclase Activity in Osteoblastic Cells,” Am. J. Physiol., 273, No. 4, Pt. 1, pp. E776–781.
Kato,  Y., Windle,  J. J., Koop,  B. A., Mundy,  G. R., and Bonewald,  L. F., 1997, “Establishment of an Osteocyte-Like Cell Line, MLO-Y4,” J. Bone Miner. Res., 12, pp. 2014–2023.
White, F. M., 1994, Fluid Mechanics, 3rd ed., McGraw-Hill, New York.
Frangos,  J. A., Eskin,  S. G., McIntire,  L. V., and Ives,  C. L., 1985, “Flow Effects on Prostacyclin Production by Cultured Human Endothelial Cells,” Science, 227, pp. 1477–1479.
Crosby,  A. H., Edwards,  S. J., Murry,  J. C., and Dixon,  M. J., 1995, “Genomic Organization of the Human Osteopontin Gene; Exclusion of the Locus From a Causative Role in the Pathogenesis of Dentinogenesis Imperfecta Type II,” Genomics, 27, pp. 155–160.
Jacobs,  C. R., Yellowley,  C. E., Nelson,  D. V., and Donahue,  H. J., 2000, “Analysis of Time-Varying Biological Data Using Rainflow Cycle Counting,” Comput. Meth. Biomech. Biomed. Eng., 3, pp. 31–40.
Brighton,  C. T., Strafford,  B., Gross,  S. B., Leatherwood,  D. F., Williams,  J. L., and Pollack,  S. R., 1991, “The Proliferative and Synthetic Response of Isolated Calvarial Bone Cells of Rats to Cyclic Biaxial Mechanical Strain,” J. Bone Jt. Surg., Am. Vol., 73, No. 3, pp. 320–331.
Geiger,  R. V., Berk,  B. C., Alexander,  R. W., and Nerem,  R. M., 1992, “FlowInduced Calcium Transients in Single Endothelial Cells: Spatial and Temporal Analysis,” Am. J. Physiol., 262, No. 6, Pt. 1, pp. C1411–1417.
Yellowley,  C. E., Jacobs,  C. R., Li,  Z., Zhou,  Z., and Donahue,  H. J., 1997, “Effects of Fluid Flow on Intracellular Calcium in Bovine Articular Chondrocytes,” Am. J. Physiol., 273, No. 1, Pt. 1, pp. C30–C36.
Knothe Tate,  M. L., Knothe,  U., and Neiderer,  P., 1998, “Experimental Elucidation of Mechanical Load-Induced Fluid Flow and Its Potential Role in Bone Metabolism and Functional Adaptation,” Am. J. Med. Sci., 316, pp. 189–195.
Weinbaum, S., Cowin, S. C., and Zeng, Y., 1991, “A Model for the Fluid Shear Stress Excitation of Membrane Ion Channels in Osteocytic Processes Due to Bone Strain,” Vanderby, R., ed., Advances in Bioengineering, ASME BED-Vol. 20.
Meldolesi,  J., and Pozzan,  T., 1987, “Pathways of Ca2+ Influx at the Plasma Membrane: Voltage-, Receptor-, and Second Messenger-Operated Channels,” Exp. Cell Res., 171, No. 2, pp. 271–283.
Dolmetsch,  R. E., Xu,  K., and Lewis,  R. S., 1998, “Calcium Oscillations Increase the Efficiency and Specificity of Gene Expression,” Nature (London), 392, No. 6679, pp. 933–936.
Horne,  J. H., 1999, “Regulatory and Spatial Aspects of Inositol Trisphosphate-Mediated Calcium Signals,” Cell Biochem. Biophys., 30, No. 2, pp. 267–286.
Ajubi,  N. E., Klein-Nulend,  J., Nijweide,  P. J., Vrijheid-Lammers,  T., Alblas,  M. J., and Burger,  E. H., 1996, “Pulsating Fluid Flow Increases Prostaglandin Production by Cultured Chicken Osteocytes—a Cytoskeleton-Dependent Process,” Biochem. Biophys. Res. Commun., 225, No. 1, pp. 62–68.
Ajubi,  N. E., Klein-Nulend,  J., Alblas,  M. J., Burger,  E. H., and Nijweide,  P. J., 1999, “Signal Transduction Pathways Involved in Fluid Flow-Induced PGE2 Production by Cultured Osteocytes,” Am. J. Physiol., 276, No. 1, Pt. 1, pp. E171–178.
Weinreb,  M., Shinar,  D., and Rodan,  G. A., 1990, “Different Pattern of Alkaline Phosphatase, Osteopontin, and Osteocalcin Expression in Developing Rat Bone Visualized by In Situ Hybridization,” J. Bone Miner. Res., 5, pp. 831–842.
Merry,  K., Dodds,  R., Littlewood,  A., and Gowen,  M., 1993, “Expression of Osteopontin mRNA by Osteoclasts and Osteoblasts in Modeling Adult Human Bone,” J. Cell. Sci., 104, pp. 1013–1020.
McKee,  M. D., Farach-Carson,  M. C., Butler,  W. T., Hauschka,  P. V., and Nanci,  A., 1993, “Ultrastructural Immunolocalization of Noncollagenous (Osteopontin and Osteocalcin) and Plasma (Albumin and α2 HS-Glycoprotein) Proteins in Rat Bone,” J. Bone Miner. Res., 8, pp. 485–496.
Yellowley,  C. E., Jacobs,  C. R, and Donahue,  H. J., 1999, “Mechanisms Contributing to Flow Induced Ca2+I Mobilization in Articular Chondrocytes,” J. Cell Physiol., 180, pp. 402–408.


Grahic Jump Location
Schematic of the substrate stretch device consisting of a silicone membrane and a computer motor-driven micrometer. One end of the membrane was fixed to the microscope stage and the other end was connected to the micrometer. Cells were cultured on the precoated membrane and a plastic ring filled with medium was placed on the membrane.
Grahic Jump Location
Membrane stretch pattern. We first induced dynamic strains of 0.1 percent for 0.5 min followed by a 3 min rest period then 1 percent strain, rest, 5 percent, rest, 10 percent, and then rest. The order of strain levels was also reversed. The strain waveform was a triangle wave and the frequency for all strain experiments was 1 Hz.
Grahic Jump Location
An example of the hFOB cell [Ca2+]i response traces obtained for oscillating flow (2 N/m2, 1 Hz). Note that the arrow depicts the onset of flow.
Grahic Jump Location
Fraction of hFOB cells responding with an increase in [Ca2+]i at different substrate strains and the mean response amplitude. 0.48±0.48 percent, 1.31±0.64 percent, 2.34±0.96 percent, 3.36±1.18 percent and 8.15±0.95 percent of hFOB cells responded for no stretch, 0.1, 1, 5, and 10 percent strain, respectively. Mean response amplitudes of hFOB cells were 37.53±6.83,45.60±10.81,41.68±14.00,39.63±6.32, and 50.57±9.91 nM for no stretch, 0.1, 1, 5, and 10 percent strain, respectively. The data were obtained from six individual experiments and a total of 246 cells. (* represents statistically significant difference (p<0.05) from other four groups, no stretch, 0.1, 1, and 5 percent strain).
Grahic Jump Location
Fraction of hFOB cells responding with an increase in [Ca2+]i at the reversing ordering of application of the various substrate strains. 1.16±0.32,9.40±1.46,4.57±0.50,3.40±0.22, and 1.90±0.29 percent of hFOB cells responded for no stretch, 10, 5, 1, and 0.1 percent strain, respectively. The data were obtained from four individual experiments and a total of 229 cells. (* represents statistically significant difference (p<0.05) from other four groups, no stretch, 5, 1, and 0.1 percent strain).
Grahic Jump Location
Fraction of cells responding with an increase in [Ca2+]i at different substrate strains for ROB cells (left) and MLO-Y4 cells (right). 0.45±0.45,0.35±0.35,2.85±0.79,3.30±0.90, and 12.46±1.91 percent were the percentages of ROB cells responding for no stretch, 0.1, 1, 5, and 10 percent strain. Only the response for 10 percent strain was significantly different from those of four other cases. The results for ROB cells were from six individual experiments that contained total 293 cells. The percentages of responding MLO-Y4 cells were 0.39±0.39,0.48±0.48,2.72±1.04,3.55±1.15, and 9.53±2.17 percent for no stretch, 0.1, 1, 5, and 10 percent strain. The response of 10 percent strain was significantly different from those for 0.1, 1, and 5 percent strain. Six individual experiments had total 227 MLO-Y4 cells. (* represents statistically significant difference (p<0.05) from other four groups, no stretch, 0.1, 1, and 5 percent strain).
Grahic Jump Location
Fraction of cells responding with an increase in [Ca2+]i to dynamic substrate strain (0.5 percent, 1 Hz) and oscillatory flow (2 N/m2, 1 Hz) for hFOB cells. The percentage numbers were 2.24±1.28 percent for dynamic strain and 12.30±1.88 percent for oscillating flow. The total cell numbers for strain and flow were 122 and 332, respectively.
Grahic Jump Location
The relative hFOB cell osteopontin mRNA level change in response to physical stimuli: substrate deformation 0.99±0.12(n=3) and oscillating fluid flow 1.95±0.40(n=2). The mRNA level was measured 72 hours after stimulation.



Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging and repositioning the boxes below.

Related Journal Articles
Related eBook Content
Topic Collections

Sorry! You do not have access to this content. For assistance or to subscribe, please contact us:

  • TELEPHONE: 1-800-843-2763 (Toll-free in the USA)
  • EMAIL: asmedigitalcollection@asme.org
Sign In