Skip to main content
NCBI home page
Cytotechnology logoLink to Cytotechnology
. 2002 Sep;39(3):125–130. doi: 10.1023/A:1023925230651

Experimental model for stimulation of cultured human osteoblast-like cells by high frequency vibration

N Rosenberg 1,2, M Levy 1, M Francis 1
PMCID: PMC3449642  PMID: 19003304

Abstract

Reliable and reproducible experimental methods for studying enhancement of osteoblast proliferation and metabolic activity in vitro provide invaluable tools for the research of biochemical processes involved in bone turnover in vivo. Some of the current methods used for this purpose are based on the ability of the osteoblasts to react metabolically to mechanical stimulation. These methods are based on the hypothesis that intracellular metabolic pathways could be influenced by the excitation of cytoskeletal components by mechanical cell deformation. Based on the same assumptions we developed a new experimental approach of biomechanical stimulation of cultured osteoblast-like cells by vibration. This method is based on the use of a specially designed vibration device that consists of an electric shaker with horizontally mounted well plate containing cell cultures. We used a first passage explant outgrowth of human osteoblast-like cell cultures, originating from samples of cancelous bone, collected from femoral necks of six donors during surgical arthroplasties of osteoarthritic hips. Well plates with replicates of cultured cells were exposed to a sine shaped vibration protocol in a frequency range of 20–60 Hz with displacement amplitude of 25 (±5) μm. We found that vibration at a distinct set of mechanical parameters of 20 Hz frequency and peak to peak acceleration of 0.5 ± 0.1 m/sec2 is optimal for cell proliferation, and at 60 Hz frequency with peak to peak acceleration of 1.3 ± 0.1 m/sec2 for metabolic activity. The presented easily reproducible experimental model should improve and simplify further research on the interactions between mechanical stimuli and intracellular biochemical pathways in osteoblasts.

Keywords: Cell culture, Mechanotransduction, Osteoblast like cells, Vibration

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

References

  1. Banes A.J., Link G.W., Gilbert J.W., Tran Son Tay R., Monbuireau O. Culturing cells in a mechanically active environment. Am Biotech Lab. 1990;8:12–22. [PubMed] [Google Scholar]
  2. Bessey O.A., Lowry O.H., Brock M.J. A method for the rapid determination of alkaline phosphatase with five cubic millimeters of serum. J Biol Chem. 1946;164:321–329. [PubMed] [Google Scholar]
  3. Boppart M.D., Kimmel D.B., Yee J.A., Cullen D.M. Time course of osteoblast appearance after in vivo mechanical loading. Bone. 1998;23:409–415. doi: 10.1016/S8756-3282(98)00119-7. [DOI] [PubMed] [Google Scholar]
  4. Buckley M.J., Banes A.J., Levin L.G., Sumpio B.E., Sato M., Jordan R., et al. Osteoblasts increase their rate of division and align in response to cyclic, mechanical tension in vitro. Bone Min. 1988;4:225–236. [PubMed] [Google Scholar]
  5. Burger E.H., Klein-Nulend J. Microgravity and bone cell mechanosensitivity. Bone. 1998;22:127S–130S. doi: 10.1016/S8756-3282(98)00010-6. [DOI] [PubMed] [Google Scholar]
  6. Duncan R.L., Turner C.H. Mechanotransduction and functional response of bone to mechanical strain. Calcif Tissue Int. 1995;57:344–358. doi: 10.1007/BF00302070. [DOI] [PubMed] [Google Scholar]
  7. Gundle R., Stewart K., Screen J., Beresford J.N. Isolation and culture of human bone-derived cells. In: Beresford N., Owen M.E., editors. Marrow Stromal Cell Culture. Cambridge, UK: Cambridge university press; 1998. pp. 43–66. [Google Scholar]
  8. Jacobs C.R., Yellowley C.E., Davis B.R., Zhou Z., Cimbala J.M., Donahue H.J. Differential effect of steady versus oscillating flow on bone cells. J Biomech. 1998;31:969–976. doi: 10.1016/S0021-9290(98)00114-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jones D.B., Nolte H., Scholubbers J.G., Turner E., Veltel D. Biochemical signal transduction of mechanical strain in osteoblast-like cells. Biomaterials. 1991;12:101–110. doi: 10.1016/0142-9612(91)90186-E. [DOI] [PubMed] [Google Scholar]
  10. Khalil A.M., Qassem W. A search for possible cytogenetic effects of low frequency vibrations in cultured human lymphocytes. Human Exp Toxicol. 1996;15:504–507. doi: 10.1177/096032719601500608. [DOI] [PubMed] [Google Scholar]
  11. Labarca C., Paigen K. A simple, rapid and sensitive DNA assay procedure. Analytical Biochem. 1980;102:344–352. doi: 10.1016/0003-2697(80)90165-7. [DOI] [PubMed] [Google Scholar]
  12. Levy M., Joyner C., Simpson A.H., Kenwright J., Francis M.J. Mechanical vibration stimulates adult human bone-derived cell proliferation in vitro. J Bone Joint Surgery 81B. 1999;Supp3:328. [Google Scholar]
  13. Liu C.C., Kalu D.N. Human parathyroid hormone-(1-34) prevents bone loss and augments bone formation in sexually mature ovariectomized rats. J Bone Min Res. 1990;5:973–982. doi: 10.1002/jbmr.5650050911. [DOI] [PubMed] [Google Scholar]
  14. Neidlinger-Wilke C., Wilke H.J., Claes L. Cyclic stretching of human osteoblasts affects proliferation and metabolism: A new experimental method and its application. J Orthop Res. 1994;12:70–78. doi: 10.1002/jor.1100120109. [DOI] [PubMed] [Google Scholar]
  15. Nigg B.M. Acceleration. In: Nigg B.M., Herzog W., editors. Biomechanics of the Musculo-Skeletal System. 2nd edn. Chichester: John Wiley & Sons; 1998. pp. 300–301. [Google Scholar]
  16. Reich K.M., McAllister T.N., Gudi S., Frangos J.A. Activation of G proteins mediates flow-induced prostaglandin E2 production in odsteoblasts. Endocrinology. 1997;138:1014–1018. doi: 10.1210/en.138.3.1014. [DOI] [PubMed] [Google Scholar]
  17. Rubin C., Li C., Sun Y., Fritton C., McLeod K. Non-invasive stimulation of trabecular bone formation via low magnitude, high frequency strain. Trans ORS. 1995;20:548. [Google Scholar]
  18. Smith E.L., Gilligan C. Dose-Response relationship between physical loading and mechanical competence of bone. Bone. 1996;18:45S–50S. doi: 10.1016/8756-3282(95)00379-7. [DOI] [PubMed] [Google Scholar]
  19. Soejima K., Klein-Nulend J., Semeins C.M., Burger E.H. Calvarial and long bone cells derived from adult mice respond similarly to pulsating fluid flow with rapid nitric oxide production. Calcif Tissue Int. 1999;64:S113. [Google Scholar]
  20. Stanford C.M., Morcuende J.A., Brand R.A. Proliferative and phenotypic responses of bone-like cells to mechanical deformation. J Orth Res. 1995;13:664–670. doi: 10.1002/jor.1100130505. [DOI] [PubMed] [Google Scholar]
  21. Tjandrawinata R.R., Vincent V.L., Hughes-Fulford M. Vibrational force alters m RNA expression in osteoblasts. FASEB. 1997;11:493–497. doi: 10.1096/fasebj.11.6.9194530. [DOI] [PubMed] [Google Scholar]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.

RESOURCES