Cellular and Molecular Bioengineering

, Volume 12, Issue 1, pp 53–67 | Cite as

Ciliotherapy Treatments to Enhance Biochemically- and Biophysically-Induced Mesenchymal Stem Cell Osteogenesis: A Comparison Study

  • M. A. Corrigan
  • T. M. Ferradaes
  • M. Riffault
  • D. A. HoeyEmail author



New approaches to treat osteoporosis have focused on promoting bone formation through the targeting of osteoblasts and their progenitors, mesenchymal stem cells (MSCs). The primary cilium is a singular cellular extension known to play an important role in biochemical and biophysical osteogenic induction of MSCs. Defects in ciliary structure have been associated with a plethora of diseases. Therefore targeting the cilium therapeutically (ciliotherapies) has emerged as a potential new treatment modality. Therefore, this study performed a comparison analysis on known ciliotherapies and their potential effects in mediating MSC osteogenic differentiation.


MSCs were treated with forskolin, lithium chloride (LiCl) or fenoldopam to investigate the effect on ciliogenesis and cilia-associated signalling. Moreover, both early and long term biochemical and biophysical (fluid shear) induced osteogenic differentiation was examined in terms of osteogenic gene expression and bone matrix deposition following each treatment.


LiCl and fenoldopam were found to enhance MSC ciliogenesis to a similar degree. LiCl significantly altered hedgehog (HH) and Wnt signalling which was associated with inhibited osteogenic gene expression, while fenoldopam demonstrated enhanced early osteogenesis. Long term treatment with both ciliotherapies did not enhance osteogenesis, however LiCl had detrimental effects on cell viability. Intriguingly both ciliotherapies enhanced MSC mechanosensitivity as demonstrated by augmented osteogenic gene expression in response to fluid shear, which over longer durations resulted in enhanced matrix deposition per cell.


Therefore, ciliotherapies can be utilised to enhance MSC ciliogenesis resulting in enhanced mechanosensitivity, however, only fenoldopam is a viable ciliotherapeutic option to enhance MSC osteogenesis.


Bone Oscillatory fluid shear Primary cilium Mechanobiology Hedgehog Wnt 



This work was supported by a European Research Council Grant 336882 (to D.A.H.); Science Foundation Ireland European Research Council (ERC) Support Grant SFI 13/ERC/L2864 (to D.A.H.).

Conflict of interest

All the authors declare that they have no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

12195_2018_561_MOESM1_ESM.docx (702 kb)
Supplementary material 1 (DOCX 702 kb)


  1. 1.
    Baron, R., and E. Hesse. Update on bone anabolics in osteoporosis treatment: rationale, current status, and perspectives. J. Clin. Endocr. Metab. 97:311–325, 2012.CrossRefGoogle Scholar
  2. 2.
    Besschetnova, T. Y., E. Kolpakova-Hart, Y. Guan, et al. Identification of signaling pathways regulating primary cilium length and flow-mediated adaptation. Curr. Biol. 20:182–187, 2010.CrossRefGoogle Scholar
  3. 3.
    Bodine, P. V. N. Wnt signaling in osteoblast differentiation and bone formation. Bone 38:S8–S8, 2006.CrossRefGoogle Scholar
  4. 4.
    Cairoli, E., C. Eller-Vainicher, F. M. Ulivieri, et al. Factors associated with bisphosphonate treatment failure in postmenopausal women with primary osteoporosis. Osteoporos. Int. 25:1401–1410, 2014.CrossRefGoogle Scholar
  5. 5.
    Chen, J. C., D. A. Hoey, M. Chua, et al. Mechanical signals promote osteogenic fate through a primary cilia-mediated mechanism. FASEB J. 30:1504–1511, 2016.CrossRefGoogle Scholar
  6. 6.
    Chen, Y., H. C. Whetstone, A. C. Lin, et al. Beta-catenin signaling plays a disparate role in different phases of fracture repair: implications for therapy to improve bone healing. PLoS Med. 4:e249, 2007.CrossRefGoogle Scholar
  7. 7.
    Corral, D. A., M. Amling, M. Priemel, et al. Dissociation between bone resorption and bone formation in osteopenic transgenic mice. Proc. Natl. Acad. Sci. U.S.A. 95:13835–13840, 1998.CrossRefGoogle Scholar
  8. 8.
    Corrigan, M. A., G. P. Johnson, E. Stavenschi, et al. TRPV4-mediates oscillatory fluid shear mechanotransduction in mesenchymal stem cells in part via the primary cilium. Sci. Rep. 8:3824, 2018.CrossRefGoogle Scholar
  9. 9.
    Curran, G., and A. Ravindran. Lithium for bipolar disorder: a review of the recent literature. Expert Rev. Neurother. 14:1079–1098, 2014.CrossRefGoogle Scholar
  10. 10.
    Dominici, M., C. Pritchard, J. E. Garlits, et al. Hematopoietic cells and osteoblasts are derived from a common marrow progenitor after bone marrow transplantation. Proc. Natl. Acad. Sci. U.S.A. 101:11761–11766, 2004.CrossRefGoogle Scholar
  11. 11.
    Downs, M. E., A. M. Nguyen, F. A. Herzog, et al. An experimental and computational analysis of primary cilia deflection under fluid flow. Comput. Methods Biomech. Biomed. Eng. 17:2–10, 2014.CrossRefGoogle Scholar
  12. 12.
    Duncan, R. L., and C. H. Turner. Mechanotransduction and the functional response of bone to mechanical strain. Calcif. Tissue Int. 57:344–358, 1995.CrossRefGoogle Scholar
  13. 13.
    Eichholz, K. F., and D. A. Hoey. Mediating human stem cell behaviour via defined fibrous architectures by melt electrospinning writing. Acta Biomater. 75:140–151, 2018.CrossRefGoogle Scholar
  14. 14.
    Evenepoel, P., P. D’haese, and V. Brandenburg. Romosozumab in postmenopausal women with osteopenia. N. Engl. J. Med. 370:1664–1664, 2014.CrossRefGoogle Scholar
  15. 15.
    Hildebrandt, F., T. Benzing, and N. Katsanis. Mechanisms of disease: ciliopathies. N. Engl. J. Med. 364:1533–1543, 2011.CrossRefGoogle Scholar
  16. 16.
    Hilgendorf, K. I., C. T. Johnson, and P. K. Jackson. The primary cilium as a cellular receiver: organizing ciliary GPCR signaling. Curr. Opin. Cell Biol. 39:84–92, 2016.CrossRefGoogle Scholar
  17. 17.
    Hoey, D. A., J. C. Chen, and C. R. Jacobs. The primary cilium as a novel extracellular sensor in bone. Front. Endocrinol. (Lausanne) 3:75, 2012.CrossRefGoogle Scholar
  18. 18.
    Hoey, D. A., M. E. Downs, and C. R. Jacobs. The mechanics of the primary cilium: an intricate structure with complex function. J. Biomech. 45:17–26, 2012.CrossRefGoogle Scholar
  19. 19.
    Hoey, D. A., S. Tormey, S. Ramcharan, et al. Primary cilia-mediated mechanotransduction in human mesenchymal stem cells. Stem Cells 30:2561–2570, 2012.CrossRefGoogle Scholar
  20. 20.
    James, A. W. Review of signaling pathways governing MSC osteogenic and adipogenic differentiation. Scientifica (Cairo) 2013:684736, 2013.Google Scholar
  21. 21.
    Jansen, K. A., D. M. Donato, H. E. Balcioglu, et al. A guide to mechanobiology: where biology and physics meet. Biochim. Biophys. Acta 1853:3043–3052, 2015.CrossRefGoogle Scholar
  22. 22.
    Johnson, G. P., E. Stavenschi, K. F. Eichholz, et al. Mesenchymal stem cell mechanotransduction is cAMP dependent and regulated by adenylyl cyclase 6 and the primary cilium. J. Cell Sci. 131:21, 2018.CrossRefGoogle Scholar
  23. 23.
    Kathem, S. H., A. M. Mohieldin, S. Abdul-Majeed, et al. Ciliotherapy: a novel intervention in polycystic kidney disease. J. Geriatr. Cardiol. 11:63–73, 2014.Google Scholar
  24. 24.
    Kreke, M. R., W. R. Huckle, and A. S. Goldstein. Fluid flow stimulates expression of osteopontin and bone sialoprotein by bone marrow stromal cells in a temporally dependent manner. Bone 36:1047–1055, 2005.CrossRefGoogle Scholar
  25. 25.
    Kulterer, B., G. Friedl, A. Jandrositz, et al. Gene expression profiling of human mesenchymal stem cells derived from bone marrow during expansion and osteoblast differentiation. BMC Genomics 8:70, 2007.CrossRefGoogle Scholar
  26. 26.
    Labour, M. N., M. Riffault, S. T. Christensen, et al. TGFbeta1-induced recruitment of human bone mesenchymal stem cells is mediated by the primary cilium in a SMAD3-dependent manner. Sci. Rep. 6:35542, 2016.CrossRefGoogle Scholar
  27. 27.
    Lee, D. J., H. C. Tseng, S. W. Wong, et al. Dopaminergic effects on in vitro osteogenesis. Bone Res. 3:15020, 2015.CrossRefGoogle Scholar
  28. 28.
    Metzger, T. A., T. C. Kreipke, T. J. Vaughan, et al. The in situ mechanics of trabecular bone marrow: the potential for mechanobiological response. J. Biomech. Eng. 65:279, 2015.CrossRefGoogle Scholar
  29. 29.
    Miyoshi, K., K. Kasahara, I. Miyazaki, et al. Factors that influence primary cilium length. Acta Med. Okayama 65:279–285, 2011.Google Scholar
  30. 30.
    Nakakura, T., A. Asano-Hoshino, T. Suzuki, et al. The elongation of primary cilia via the acetylation of alpha-tubulin by the treatment with lithium chloride in human fibroblast KD cells. Med. Mol. Morphol. 48:44–53, 2015.CrossRefGoogle Scholar
  31. 31.
    Nakamura, T., M. Naruse, Y. Chiba, et al. Novel hedgehog agonists promote osteoblast differentiation in mesenchymal stem cells. J. Cell. Physiol. 230:922–929, 2015.CrossRefGoogle Scholar
  32. 32.
    Ou, Y., Y. Ruan, M. Cheng, et al. Adenylate cyclase regulates elongation of mammalian primary cilia. Exp. Cell Res. 315:2802–2817, 2009.CrossRefGoogle Scholar
  33. 33.
    Resnick, A. Mechanical properties of a primary cilium as measured by resonant oscillation. Biophys. J. 109:18–25, 2015.CrossRefGoogle Scholar
  34. 34.
    Rowson, D., M. M. Knight, and H. R. Screen. Zonal variation in primary cilia elongation correlates with localized biomechanical degradation in stress deprived tendon. J. Orthop. Res. 34:2146–2153, 2016.CrossRefGoogle Scholar
  35. 35.
    Salek, M. M., P. Sattari, and R. J. Martinuzzi. Analysis of fluid flow and wall shear stress patterns inside partially filled agitated culture well plates. Ann. Biomed. Eng. 40:707–728, 2012.CrossRefGoogle Scholar
  36. 36.
    Sharma, N., Z. A. Kosan, J. E. Stallworth, et al. Soluble levels of cytosolic tubulin regulate ciliary length control. Mol. Biol. Cell 22:806–816, 2011.CrossRefGoogle Scholar
  37. 37.
    Singla, V., and J. F. Reiter. The primary cilium as the cell’s antenna: signaling at a sensory organelle. Science 313:629–633, 2006.CrossRefGoogle Scholar
  38. 38.
    Spasic, M., and C. R. Jacobs. Lengthening primary cilia enhances cellular mechanosensitivity. Eur. Cell Mater. 33:158–168, 2017.CrossRefGoogle Scholar
  39. 39.
    Stavenschi, E., M. N. Labour, and D. A. Hoey. Oscillatory fluid flow induces the osteogenic lineage commitment of mesenchymal stem cells: the effect of shear stress magnitude, frequency, and duration. J. Biomech. 55:99–106, 2017.CrossRefGoogle Scholar
  40. 40.
    Thompson, C. L., A. Wiles, C. A. Poole, et al. Lithium chloride modulates chondrocyte primary cilia and inhibits hedgehog signaling. FASEB J. 30:716–726, 2016.CrossRefGoogle Scholar
  41. 41.
    Tummala, P., E. J. Arnsdorf, and C. R. Jacobs. The role of primary cilia in mesenchymal stem cell differentiation: a pivotal switch in guiding lineage commitment. Cell. Mol. Bioeng. 3:207–212, 2010.CrossRefGoogle Scholar
  42. 42.
    Tuson, M., M. He, and K. V. Anderson. Protein kinase A acts at the basal body of the primary cilium to prevent Gli2 activation and ventralization of the mouse neural tube. Development 138:4921–4930, 2011.CrossRefGoogle Scholar
  43. 43.
    Upadhyay, V. S., B. S. Muntean, S. H. Kathem, et al. Roles of dopamine receptor on chemosensory and mechanosensory primary cilia in renal epithelial cells. Front. Physiol. 5:72, 2014.CrossRefGoogle Scholar
  44. 44.
    Yang, Y. J., J. Z. Yang, R. X. Liu, et al. Accumulation of beta-catenin by lithium chloride in porcine myoblast cultures accelerates cell differentiation. Mol. Biol. Rep. 38:2043–2049, 2011.CrossRefGoogle Scholar
  45. 45.
    Zamani, A., G. R. Omrani, and M. M. Nasab. Lithium’s effect on bone mineral density. Bone 44:331–334, 2009.CrossRefGoogle Scholar
  46. 46.
    Zhang, X., E. M. Schwarz, D. A. Young, et al. Cyclooxygenase-2 regulates mesenchymal cell differentiation into the osteoblast lineage and is critically involved in bone repair. J. Clin. Invest. 109:1405–1415, 2002.CrossRefGoogle Scholar
  47. 47.
    Zhu, Z., J. Yin, J. Guan, et al. Lithium stimulates human bone marrow derived mesenchymal stem cell proliferation through GSK-3beta-dependent beta-catenin/Wnt pathway activation. FEBS J. 281:5371–5389, 2014.CrossRefGoogle Scholar

Copyright information

© Biomedical Engineering Society 2018

Authors and Affiliations

  1. 1.Trinity Centre for Bioengineering, Trinity Biomedical Sciences InstituteTrinity CollegeDublinIreland
  2. 2.Dept. of Mechanical and Manufacturing Engineering, School of EngineeringTrinity College DublinDublin 2Ireland
  3. 3.School of EngineeringUniversidade Federal do Estado do Rio de JaneiroRio do JaneiroBrazil
  4. 4.Advanced Materials and Bioengineering Research CentreTrinity College Dublin & RCSIDublin 2Ireland

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