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Cyclic hydrostatic compression stimulates chondroinduction of C3H/10T1/2 cells

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Abstract

While the potential for intermittent hydrostatic pressure to promote cartilaginous matrix synthesis is well established, its potential to influence chondroinduction remains poorly understood. This study examined the effects of relatively short- and long-duration cyclic hydrostatic compression on the chondroinduction of C3H/10T1/2 murine embryonic fibroblasts by recombinant human bone morphogenetic protein-2 (rhBMP-2). Cells were seeded at high density into round bottom wells of a 96-well plate and supplemented with 25 ng/ml rhBMP-2. Experimental cultures were subjected to either 1,800 cycles/day or 7,200 cycles/day of 1 Hz sinusoidal hydrostatic compression to 5 MPa (applied 10 min on/10 min off) for 3 days. Non-pressurized control and experimental cultures were maintained in static culture for an additional 5 days. Cultures were then analyzed for alcian blue staining intensity, DNA and sulfated glycosaminoglycan (sGAG) content, and for the rate of collagen synthesis. Whereas cultures subjected to 1,800 pressure cycles exhibited no significant differences (statistical or qualitative) compared to controls, those subjected to 7,200 cycles stained more intensely with alcian blue, contained nearly twice as much sGAG, and displayed twice the rate of collagen synthesis as non-pressurized controls. This study demonstrates the potential for cyclic hydrostatic compression to stimulate chondrogenic differentiation of the C3H/10T1/2 cell line in a duration-dependent manner.

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References

  • Angele P, Yoo JU, Smith C, Mansour J, Jepsen KJ, Nerlich M, Johnstone B (2003) Cyclic hydrostatic pressure enhances the chondrogenic phenotype of human mesenchymal progenitor cells differentiated in vitro. J Orthop Res 21(3):451–457

    Google Scholar 

  • Ashhurst DE (1986) The influence of mechanical conditions on the healing of experimental fractures in the rabbit: a microscopical study. Philos Trans R Soc Lond B Biol Sci 313(1161):271–302

    Google Scholar 

  • Atkinson BL, Fantle KS, Benedict JJ, Huffer WE, Gutierrez-Hartmann A (1997) Combination of osteoinductive bone proteins differentiates mesenchymal C3H/10T1/2 cells specifically to the cartilage lineage. J Cell Biochem. 65(3):325–339

    Google Scholar 

  • Bachner D, Schroder D, Betat N, Ahrens M, Gross G (1999) Apolipoprotein E (ApoE), a Bmp-2 (bone morphogenetic protein) upregulated gene in mesenchymal progenitors (C3H10T1/2), is highly expressed in murine embryonic development. Biofactors 9(1):11–17

    Google Scholar 

  • Bachrach NM, Valhmu WB, Stazzone E, Ratcliffe A, Lai WM, Mow VC (1995) Changes in proteoglycan synthesis of chondrocytes in articular cartilage are associated with the time-dependent changes in their mechanical environment. J Biomech 28(12):1561–1569

    Google Scholar 

  • Bartsch JW, Jackel M, Perz A, Jockusch H. (2000) Steroid RU 486 inducible myogenesis by 10T1/2 fibroblastic mouse cells. FEBS Lett 467(1):123–127

    Google Scholar 

  • Bassett CAL, Herrmann I (1961) Influence of oxygen concentration and mechanical factors on differentiation of connective tissue in vitro. Nature 190(4774):460–461

    Google Scholar 

  • Carter DR, Orr TE, Fyhrie DP, Schurman DJ (1987) Influences of mechanical stress on prenatal and postnatal skeletal development. Clin Orthop 219:237–250

    Google Scholar 

  • Carter DR, Blenman PR, Beaupre GS (1988) Correlations between mechanical stress history and tissue differentiation in initial fracture healing. J Orthop Res 6(5):736–748

    Google Scholar 

  • Carver SE, Heath CA (1999a) Increasing extracellular matrix production in regenerating cartilage with intermittent physiological pressure. Biotechnol Bioeng 62(2):166–174

    Google Scholar 

  • Carver SE, Heath CA (1999b) Semi-continuous perfusion system for delivering intermittent physiological pressure to regenerating cartilage. Tissue Eng 5(1):1–11

    Google Scholar 

  • Chen TL, Shen WJ, Kraemer FB (2001) Human BMP-7/OP-1 induces the growth and differentiation of adipocytes and osteoblasts in bone marrow stromal cell cultures. J Cell Biochem 82(2):187–199

    Google Scholar 

  • D’Andrea P, Calabrese A, Capozzi I, Grandolfo M, Tonon R, Vittur F (2000) Intercellular Ca2+ waves in mechanically stimulated articular chondrocytes. Biorheology 37(1–2):75–83

    Google Scholar 

  • Denker AE, Haas AR, Nicoll SB, Tuan RS (1999) Chondrogenic differentiation of murine C3H10T1/2 multipotential mesenchymal cells: I. Stimulation by bone morphogenetic protein-2 in high-density micromass cultures. Differentiation 64(2):67–76

    Google Scholar 

  • Elder SH, Kimura JH, Soslowsky LJ, Lavagnino M, Goldstein SA (2000) Effect of compressive loading on chondrocyte differentiation in agarose cultures of chick limb-bud cells. J Orthop Res 18(1):78–86

    Google Scholar 

  • Ellison BE, Carter DR, Smith RL (1994) Effect of variable duration intermittent hydrostatic pressure on cartilage glycoaminoglycan synthesis. Trans Orthop Res Soc 19:486

    Google Scholar 

  • Hall AC, Urban JPG, Gehl KA (1991) The effects of hydrostatic pressure on matrix synthesis in articular cartilage. J Orthop Res 9:1–10

    Google Scholar 

  • Hollnagel A, Ahrens M, Gross G (1997) Parathyroid hormone enhances early and suppresses late stages of osteogenic and chondrogenic development in a BMP-dependent mesenchymal differentiation system (C3H10T1/2). J Bone Miner Res 12(12):1993–2004

    Google Scholar 

  • Ikenoue T, Trindade MC, Lee MS, Lin EY, Schurman DJ, Goodman SB, Smith RL (2003) Mechanoregulation of human articular chondrocyte aggrecan and type II collagen expression by intermittent hydrostatic pressure in vitro. J Orthop Res 21(1):110–116

    Google Scholar 

  • Yusuhiko K, Hugo H, Juergen M (1997) Analysis of 3H-Proline-labeled protein by rapid filtration in multiwell plates for the study of collagen metabolism. Biotechniques 22:706–716

    Google Scholar 

  • Mauck RL, Oswald ES, Cheng Q, Majumdar MK, Nicoll SB, Ateshian GA, Hung CT (2003) Hydrostatic pressure enhances chondrogenic differentiation of human multipotential mesenchymal cells in alginate disks. In: Trans ASME summer bioengineering conference, pp 265–266

  • Mow VC, Ratcliffe A, Poole AR (1992) Cartilage and diarthrodial joints as paradigms for hierarchical materials and structures. Biomaterials 13(2):67–97

    Google Scholar 

  • Nagatomi J, Arulanandam BP, Metzger DW, Meunier A, Bizios R (2001) Frequency- and duration-dependent effects of cyclic pressure on select bone cell functions. Tissue Eng 7(6):717–728

    Google Scholar 

  • Page M, Ashhurst DE (1987) The effects of mechanical stability on the macromolecules of the connective tissue matrices produced during fracture healing. II. The glycosaminoglycans. Histochem J 19(1):39–61

    Google Scholar 

  • Page M, Hogg J, Ashhurst DE (1986) The effects of mechanical stability on the macromolecules of the connective tissue matrices produced during fracture healing. I. The collagens. Histochem J 18(5):251–265

    CAS  PubMed  Google Scholar 

  • Parkkinen JJ, Ikonen J, Lammi MJ, Laakkonen J, Tammi M, Helminen HJ (1993) Effects of cyclic hydrostatic pressure on proteoglycan synthesis in cultured chondrocytes and articular cartilage explants. Arch Biochem Biophys 300(1):458–465

    Google Scholar 

  • Pauwels F (1980) A new theory concerning the influence of mechanical stimuli on the differentiation of the supporting tissues. In: Biomechanics of the locomotor apparatus: contributions on the functional anatomy of the locomotor apparatus. Springer, Berlin Heidelberg New York, pp 375–407

    Google Scholar 

  • Seghatoleslami MR, Roman-Blas JA, Rainville AM, Modaressi R, Danielson KG, Tuan RS (2003) Progression of chondrogenesis in C3H10T1/2 cells is associated with prolonged and tight regulation of ERK1/2. J Cell Biochem 88(6):1129–1144

    Google Scholar 

  • Singh R, Artaza JN, Taylor WE, Gonzalez-Cadavid NF, Bhasin S (2003) Androgens stimulate myogenic differentiation and inhibit adipogenesis in C3H 10T1/2 pluripotent cells through an androgen receptor-mediated pathway. Endocrinology 144(11):5081–5088

    Google Scholar 

  • Smith RL, Lin J, Trindade MC, Shida J, Kajiyama G, Vu T, Hoffman AR, van der Meulen MC, Goodman SB, Schurman DJ, Carter DR (2000) Time-dependent effects of intermittent hydrostatic pressure on articular chondrocyte type II collagen and aggrecan mRNA expression. J Rehabil Res Dev 37(2):153–161

    Google Scholar 

  • Spinella-Jaegle S, Rawadi G, Kawai S, Gallea S, Faucheu C, Mollat P, Courtois B, Bergaud B, Ramez V, Blanchet AM, Adelmant G, Baron R, Roman-Roman S (2001) Sonic hedgehog increases the commitment of pluripotent mesenchymal cells into the osteoblastic lineage and abolishes adipocytic differentiation. J Cell Sci 114(Pt 11):2085–2094

    Google Scholar 

  • Takahashi I, Nuckolls GH, Takahashi K, Tanaka O, Semba I, Dashner R, Shum L, Slavkin HC (1998) Compressive force promotes sox9, type II collagen and aggrecan and inhibits IL-1beta expression resulting in chondrogenesis in mouse embryonic limb bud mesenchymal cells. J Cell Sci 111(Pt 14):2067–2076

    Google Scholar 

  • Tang QQ, Otto TC, Lane MD (2004) Commitment of C3H10T1/2 pluripotent stem cells to the adipocyte lineage. Proc Natl Acad Sci USA 101(26):9607–9611

    Google Scholar 

  • Taylor SM, Jones PA (1979) Multiple new phenotypes induced in 10T1/2 and 3T3 cells treated with 5-azacytidine. Cell 17(4):771–779

    Google Scholar 

  • Toyoda T, Seedhom BB, Kirkham J, Bonass WA. (2003) Upregulation of aggrecan and type II collagen mRNA expression in bovine chondrocytes by the application of hydrostatic pressure. Biorheology 40(1–3):79–85

    Google Scholar 

  • Wong M, Siegrist M, Goodwin K, Park Y (2002) Hydrostatic pressure, tension and unconfined compression differentially regulate expression of cartilage matrix proteins. Trans Orthop Res Soc 27:33

    Google Scholar 

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Acknowledgements

This research was supported by a Whitaker Foundation Biomedical Engineering Research Grant (RG-00-0371) and the Department of Agricultural & Biological Engineering, Mississippi State University. rhBMP-2 was a generous gift of Wyeth Pharmaceuticals, Cambridge, MA. The authors would also like to thank Dr. Randal Buddington, Professor of Biological Sciences at Mississippi State University, for his technical assistance.

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Correspondence to SH Elder.

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Elder, S., Fulzele, K. & McCulley, W. Cyclic hydrostatic compression stimulates chondroinduction of C3H/10T1/2 cells. Biomech Model Mechanobiol 3, 141–146 (2005). https://doi.org/10.1007/s10237-004-0058-3

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  • DOI: https://doi.org/10.1007/s10237-004-0058-3

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