Skip to main content

Advertisement

SpringerLink
Log in

Extracellular Matrix Produced by Osteoblasts Cultured Under Low-Magnitude, High-Frequency Stimulation is Favourable to Osteogenic Differentiation of Mesenchymal Stem Cells

  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Abstract

The effects of low-magnitude, high-frequency (LMHF) mechanical stimulation on osteoblastic cells are poorly understood. We have developed a system that generates very small (15–40 με), high-frequency (400 Hz, sine) deformations on osteoblast cultures (MC3T3-E1). We investigated the effects of these LMHF stimulations mainly on extracellular matrix (ECM) synthesis. The functional properties of this ECM after decellularization were evaluated on C3H10T1/2 mesenchymal stem cells (MSCs). LMHF stimulations were applied 20 min once daily for 1, 3, or 7 days in MC3T3-E1 culture (1, 3, or 7 dLMHF). Cell number and viability were not affected after 3 or 7 dLMHF. Osteoblast response to LMHF was assessed by an increase in nitric oxide secretion, alteration of the cytoskeleton, and focal contacts. mRNA expression for fibronectin, osteopontin, bone sialoprotein, and type I collagen in LMHF cultures were 1.8-, 1.6-, 1.5-, and 1.7-fold higher than controls, respectively (P < 0.05). In terms of protein, osteopontin levels were increased after 3 dLMHF and ECM organization was altered as shown by fibronectin topology after 7 dLMHF. After decellularization, 7 dLMHF-ECM or control ECM was reseeded with MSCs. Seven dLMHF-ECM improved early events such as cell attachment (2 h) and focal contact adhesion (6 h) and, later (16 h), modified MSC morphological parameters. After 5 days in multipotential medium, gene-expression changes indicated that 7 dLMHF-ECM promoted the expression of osteoblast markers at the expense of adipogenic marker. LMHF stimulations of osteoblasts are therefore efficient and sufficient to generate osteogenic matrix.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Frost HM (2003) Bone’s mechanostat: a 2003 update. Anat Rec A Discov Mol Cell Evol Biol 275:1081–1101

    Article  PubMed  Google Scholar 

  2. Torcasio A, van Lenthe GH, Van Oosterwyck H (2008) The importance of loading frequency, rate and vibration for enhancing bone adaptation and implant osseointegration. Eur Cell Mater 16:56–68

    PubMed  CAS  Google Scholar 

  3. Fritton SP, McLeod KJ, Rubin CT (2000) Quantifying the strain history of bone: spatial uniformity and self-similarity of low-magnitude strains. J Biomech 33:317–325

    Article  PubMed  CAS  Google Scholar 

  4. Rubin C, Turner AS, Bain S, Mallinckrodt C, McLeod K (2001) Anabolism, low mechanical signals strengthen long bones. Nature 412:603–604

    Article  PubMed  CAS  Google Scholar 

  5. Judex S, Boyd S, Qin YX, Turner S, Ye K, Muller R, Rubin C (2003) Adaptations of trabecular bone to low magnitude vibrations result in more uniform stress and strain under load. Ann Biomed Eng 31:12–20

    Article  PubMed  Google Scholar 

  6. Qin YX, Rubin CT et al (1998) Nonlinear dependence of loading intensity and cycle number in the maintenance of bone mass and morphology. J Orthop Res 16:482–489

    Article  PubMed  CAS  Google Scholar 

  7. Rubin C, Recker R, Cullen D, Ryaby J, McCabe J, McLeod K (2004) Prevention of postmenopausal bone loss by a low-magnitude, high-frequency mechanical stimuli: a clinical trial assessing compliance, efficacy, and safety. J Bone Miner Res 19:343–351

    Article  PubMed  Google Scholar 

  8. Verschueren SM, Roelants M, Delecluse C, Swinnen S, Vanderschueren D, Boonen S (2004) Effect of 6-month whole body vibration training on hip density, muscle strength, and postural control in postmenopausal women: a randomized controlled pilot study. J Bone Miner Res 19:352–359

    Article  PubMed  Google Scholar 

  9. Prisby RD, Lafage-Proust MH, Malaval L, Belli A, Vico L (2008) Effects of whole body vibration on the skeleton and other organ systems in man and animal models: what we know and what we need to know. Ageing Res Rev 7:319–329

    Article  PubMed  Google Scholar 

  10. Bacabac RG, Van Loon JJ, Smit TH, Klein-Nulend J (2009) Noise enhances the rapid nitric oxide production by bone cells in response to fluid shear stress. Technol Health Care 17:57–65

    PubMed  Google Scholar 

  11. Bacabac RG, Smit TH, Van Loon JJ, Doulabi BZ, Helder M, Klein-Nulend J (2006) Bone cell responses to high-frequency vibration stress: does the nucleus oscillate within the cytoplasm? FASEB J 20:858–864

    Article  PubMed  CAS  Google Scholar 

  12. Salgado AJ, Coutinho OP, Reis RL (2004) Bone tissue engineering: state of the art and future trends. Macromol Biosci 4:743–765

    Article  PubMed  CAS  Google Scholar 

  13. Cukierman E, Pankov R, Stevens DR, Yamada KM (2001) Taking cell-matrix adhesions to the third dimension. Science 294:1708–1712

    Article  PubMed  CAS  Google Scholar 

  14. Salasznyk RM, Williams WA, Boskey A, Batorsky A, Plopper GE (2004) Adhesion to vitronectin and collagen I promotes osteogenic differentiation of human mesenchymal stem cells. J Biomed Biotechnol 2004:24–34

    Article  PubMed  Google Scholar 

  15. Mao Y, Schwarzbauer JE (2005) Stimulatory effects of a three-dimensional microenvironment on cell-mediated fibronectin fibrillogenesis. J Cell Sci 118:4427–4436

    Article  PubMed  CAS  Google Scholar 

  16. Usson Y, Guignandon A, Laroche N, Lafage-Proust MH, Vico L (1997) Quantitation of cell-matrix adhesion using confocal image analysis of focal contact associated proteins and interference reflection microscopy. Cytometry 28:298–304

    Article  PubMed  CAS  Google Scholar 

  17. Turner CH, Forwood MR, Rho JY, Yoshikawa T (1994) Mechanical loading thresholds for lamellar and woven bone formation. J Bone Miner Res 9:87–97

    Article  PubMed  CAS  Google Scholar 

  18. Brown TD (2000) Techniques for mechanical stimulation of cells in vitro: a review. J Biomech 33:3–14

    Article  PubMed  CAS  Google Scholar 

  19. Tanaka SM (1999) A new mechanical stimulator for cultured bone cells using piezoelectric actuator. J Biomech 32:427–430

    Article  PubMed  CAS  Google Scholar 

  20. Sato K, Adachi T, Matsuo M, Tomita Y (2005) Quantitative evaluation of threshold fiber strain that induces reorganization of cytoskeletal actin fiber structure in osteoblastic cells. J Biomech 38:1895–1901

    Article  PubMed  Google Scholar 

  21. Yang RS, Lin WL, Chen YZ, Tang CH, Huang TH, Lu BY, Fu WM (2005) Regulation by ultrasound treatment on the integrin expression and differentiation of osteoblasts. Bone 36:276–283

    Article  PubMed  CAS  Google Scholar 

  22. Mierke CT (2009) The role of vinculin in the regulation of the mechanical properties of cells. Cell Biochem Biophys 53:115–126

    Article  PubMed  CAS  Google Scholar 

  23. Guignandon A, Akhouayri O, Usson Y, Rattner A, Laroche N, Lafage-Proust MH, Alexandre C, Vico L (2003) Focal contact clustering in osteoblastic cells under mechanical stresses: microgravity and cyclic deformation. Cell Commun Adhes 10:69–83

    Article  PubMed  Google Scholar 

  24. Jackson RA, Kumarasuriyar A, Nurcombe V, Cool SM (2006) Long-term loading inhibits ERK1/2 phosphorylation and increases FGFR3 expression in MC3T3-E1 osteoblast cells. J Cell Physiol 209:894–904

    Article  PubMed  CAS  Google Scholar 

  25. Tang L, Lin Z, Li YM (2006) Effects of different magnitudes of mechanical strain on osteoblasts in vitro. Biochem Biophys Res Commun 344:122–128

    Article  PubMed  CAS  Google Scholar 

  26. Rath B, Nam J, Knobloch TJ, Lannutti JJ, Agarwal S (2008) Compressive forces induce osteogenic gene expression in calvarial osteoblasts. J Biomech 41:1095–1103

    Article  PubMed  Google Scholar 

  27. Ignatius A, Blessing H, Liedert A, Schmidt C, Neidlinger-Wilke C, Kaspar D, Friemert B, Claes L (2005) Tissue engineering of bone: effects of mechanical strain on osteoblastic cells in type I collagen matrices. Biomaterials 26:311–318

    Article  PubMed  CAS  Google Scholar 

  28. Di Palma F, Guignandon A, Chamson A, Lafage-Proust MH, Laroche N, Peyroche S, Vico L, Rattner A (2005) Modulation of the responses of human osteoblast-like cells to physiologic mechanical strains by biomaterial surfaces. Biomaterials 26:4249–4257

    Article  PubMed  CAS  Google Scholar 

  29. Zhao L, Li G, Chan KM, Wang Y, Tang PF (2009) Comparison of multipotent differentiation potentials of murine primary bone marrow stromal cells and mesenchymal stem cell line C3H10T1/2. Calcif Tissue Int 84:56–64

    Article  PubMed  CAS  Google Scholar 

  30. Buckbinder L, Crawford DT, Qi H, Ke HZ, Olson LM, Long KR, Bonnette PC, Baumann AP, Hambor JE, Grasser WA 3rd, Pan LC, Owen TA, Luzzio MJ, Hulford CA, Gebhard DF, Paralkar VM, Simmons HA, Kath JC, Roberts WG, Smock SL, Guzman-Perez A, Brown TA, Li M (2007) Proline-rich tyrosine kinase 2 regulates osteoprogenitor cells and bone formation, and offers an anabolic treatment approach for osteoporosis. Proc Natl Acad Sci USA 104:10619–10624

    Article  PubMed  CAS  Google Scholar 

  31. Kim JB, Leucht P, Luppen CA, Park YJ, Beggs HE, Damsky CH, Helms JA (2007) Reconciling the roles of FAK in osteoblast differentiation, osteoclast remodeling, and bone regeneration. Bone 41:39–51

    Article  PubMed  CAS  Google Scholar 

  32. Garcia AJ, Keselowsky BG (2002) Biomimetic surfaces for control of cell adhesion to facilitate bone formation. Crit Rev Eukaryot Gene Expr 12:151–162

    Article  PubMed  CAS  Google Scholar 

  33. Roessler S, Born R, Scharnweber D, Worch H, Sewing A, Dard M (2001) Biomimetic coatings functionalized with adhesion peptides for dental implants. J Mater Sci Mater Med 12:871–877

    Article  PubMed  CAS  Google Scholar 

  34. McBeath R, Pirone DM, Nelson CM, Bhadriraju K, Chen CS (2004) Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell 6:483–495

    Article  PubMed  CAS  Google Scholar 

  35. Koutnikova H, Auwerx J (2001) Regulation of adipocyte differentiation. Ann Med 33:556–561

    Article  PubMed  CAS  Google Scholar 

  36. Puleo DA, Nanci A (1999) Understanding and controlling the bone–implant interface. Biomaterials 20:2311–2321

    Article  PubMed  CAS  Google Scholar 

  37. Bernards MT, Qin C, Ratner BD, Jiang S (2008) Adhesion of MC3T3-E1 cells to bone sialoprotein and bone osteopontin specifically bound to collagen I. J Biomed Mater Res A 86:779–787

    PubMed  Google Scholar 

  38. Bornstein P, Sage EH (2002) Matricellular proteins: extracellular modulators of cell function. Curr Opin Cell Biol 14:608–616

    Article  PubMed  CAS  Google Scholar 

  39. Mukherjee BB, Nemir M, Beninati S, Cordella-Miele E, Singh K, Chackalaparampil I, Shanmugam V, DeVouge MW, Mukherjee AB (1995) Interaction of osteopontin with fibronectin and other extracellular matrix molecules. Ann NY Acad Sci 760:201–212

    Article  PubMed  CAS  Google Scholar 

  40. Chen Y, Bal BS, Gorski JP (1992) Calcium and collagen binding properties of osteopontin, bone sialoprotein, and bone acidic glycoprotein-75 from bone. J Biol Chem 267:24871–24878

    PubMed  CAS  Google Scholar 

  41. Nakamura M, Sone S, Takahashi I, Mizoguchi I, Echigo S, Sasano Y (2005) Expression of versican and ADAMTS1, 4, and 5 during bone development in the rat mandible and hind limb. J Histochem Cytochem 53:1553–1562

    Article  PubMed  CAS  Google Scholar 

  42. Sasano Y, Li HC, Zhu JX, Imanaka-Yoshida K, Mizoguchi I, Kagayama M (2000) Immunohistochemical localization of type I collagen, fibronectin and tenascin C during embryonic osteogenesis in the dentary of mandibles and tibias in rats. Histochem J 32:591–598

    Article  PubMed  CAS  Google Scholar 

  43. Stephansson SN, Byers BA, Garcia AJ (2002) Enhanced expression of the osteoblastic phenotype on substrates that modulate fibronectin conformation and integrin receptor binding. Biomaterials 23:2527–2534

    Article  PubMed  CAS  Google Scholar 

  44. Keselowsky BG, Collard DM, Garcia AJ (2005) Integrin binding specificity regulates biomaterial surface chemistry effects on cell differentiation. Proc Natl Acad Sci USA 102:5953–5957

    Article  PubMed  CAS  Google Scholar 

  45. Martino MM, Mochizuki M, Rothenfluh DA, Rempel SA, Hubbell JA, Barker TH (2009) Controlling integrin specificity and stem cell differentiation in 2D and 3D environments through regulation of fibronectin domain stability. Biomaterials 30:1089–1097

    Article  PubMed  CAS  Google Scholar 

  46. Muruganandan S, Roman AA, Sinal CJ (2009) Adipocyte differentiation of bone marrow-derived mesenchymal stem cells: cross talk with the osteoblastogenic program. Cell Mol Life Sci 66:236–253

    Article  PubMed  CAS  Google Scholar 

  47. David V, Martin A, Lafage-Proust MH, Malaval L, Peyroche S, Jones DB, Vico L, Guignandon A (2007) Mechanical loading down-regulates peroxisome proliferator-activated receptor gamma in bone marrow stromal cells and favors osteoblastogenesis at the expense of adipogenesis. Endocrinology 148:2553–2562

    Article  PubMed  CAS  Google Scholar 

  48. Pregizer S, Baniwal SK, Yan X, Borok Z, Frenkel B (2008) Progressive recruitment of Runx2 to genomic targets despite decreasing expression during osteoblast differentiation. J Cell Biochem 105:965–970

    Article  PubMed  CAS  Google Scholar 

  49. Klees RF, Salasznyk RM, Kingsley K, Williams WA, Boskey A, Plopper GE (2005) Laminin-5 induces osteogenic gene expression in human mesenchymal stem cells through an ERK-dependent pathway. Mol Biol Cell 16:881–890

    Article  PubMed  CAS  Google Scholar 

  50. Dalby MJ, Gadegaard N, Curtis AS, Oreffo RO (2007) Nanotopographical control of human osteoprogenitor differentiation. Curr Stem Cell Res Ther 2:129–138

    Article  PubMed  CAS  Google Scholar 

  51. Dalby MJ, Gadegaard N, Tare R, Andar A, Riehle MO, Herzyk P, Wilkinson CD, Oreffo RO (2007) The control of human mesenchymal cell differentiation using nanoscale symmetry and disorder. Nat Mater 6:997–1003

    Article  PubMed  CAS  Google Scholar 

  52. Hu J, Liu X, Ma PX (2008) Induction of osteoblast differentiation phenotype on poly(l-lactic acid) nanofibrous matrix. Biomaterials 29:3815–3821

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

V. D. held a research fellowship from La Region Rhone Alpes.

Author information

Authors and Affiliations

  1. Université de Lyon, 42023, Saint-Etienne, France

    Virginie Dumas, Anthony Perrier, Carole Fournier, Alain Guignandon, Mireille Thomas, Sylvie Peyroche, Laurence Vico & Aline Rattner

  2. Laboratoire de Biologie du Tissu Osseux, IFR143, INSERM U890, 42023, Saint-Etienne, France

    Virginie Dumas, Anthony Perrier, Carole Fournier, Alain Guignandon, Mireille Thomas, Sylvie Peyroche, Laurence Vico & Aline Rattner

  3. Laboratoire de Génie Electrique et Ferroélectricité, Institut National des Sciences Appliquées de Lyon, Lyon, France

    Benjamin Ducharne & Daniel Guyomar

Authors
  1. Virginie Dumas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Benjamin Ducharne
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anthony Perrier
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Carole Fournier
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alain Guignandon
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mireille Thomas
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Sylvie Peyroche
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Daniel Guyomar
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Laurence Vico
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Aline Rattner
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Aline Rattner.

Additional information

The authors have stated that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dumas, V., Ducharne, B., Perrier, A. et al. Extracellular Matrix Produced by Osteoblasts Cultured Under Low-Magnitude, High-Frequency Stimulation is Favourable to Osteogenic Differentiation of Mesenchymal Stem Cells. Calcif Tissue Int 87, 351–364 (2010). https://doi.org/10.1007/s00223-010-9394-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00223-010-9394-8

Keywords

Search

Navigation