Skip to main content

Advertisement

SpringerLink
Log in

In vitro evaluation of bioactive strontium-based ceramic with rabbit adipose-derived stem cells for bone tissue regeneration

  • Published:
Journal of Materials Science: Materials in Medicine Aims and scope Submit manuscript

Abstract

The development of bone replacement materials is an important objective in the field of orthopaedic surgery. Due to the drawbacks of treating bone defects with autografts, synthetic bone graft materials have become optional. So in this work, a bone tissue engineering approach with radiopaque bioactive strontium incorporated calcium phosphate was proposed for the preliminary cytocompatibility studies for bone substitutes. Accumulating evidence indicates that strontium containing biomaterials promote enhanced bone repair and radiopacity for easy imaging. Hence, strontium calcium phosphate (SrCaPO4) and hydroxyapatite scaffolds have been investigated for its ability to support and sustain the growth of rabbit adipose-derived mesenchymal stem cells (RADMSCs) in vitro. They were characterized via Micro-CT for pore size distribution. Cells used were isolated from New Zealand White rabbit adipose tissue, characterized by FACS and via differentiation into the osteogenic lineage by alkaline phosphatase, Masson’s trichome, Alizarin Red and von Kossa staining on day 28. Material-cell interaction was observed by SEM imaging of cell morphology on contact with material. Live–Dead analysis was done by confocal laser scanning microscopy and cell cluster analysis via μCT. The in vitro biodegradation, elution and nucleation of apatite formation of the material was evaluated using simulated body fluid and phosphate buffered saline in static regime up to 28 days at 37 °C. These results demonstrated that SrCaPO4 is a good candidate for bone tissue engineering applications and with osteogenically-induced RADMSCs, they may serve as potential implants for the repair of critical-sized bone defects.

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
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12

Similar content being viewed by others

References

  1. Langer R, Vacanti JP. Tissue engineering. Science. 1993;14:920–6.

    Article  Google Scholar 

  2. Klara T, Csonge L, Janositz G, Csernatony Z, Lacaza. Albumin-coated structural lyophilized bone allografts: a clinical report of 10 cases. Cell Tissue Bank 2013 (in press).

  3. Giannoudis PV, Einhorn TA, Marsh D. Fracture healing: the diamond concept. Injury. 2007;4:3–6.

    Article  Google Scholar 

  4. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 2001;2:211–28.

    Article  Google Scholar 

  5. Kakudo N, Shimotsuma A, Miyake S, Kushida S, Kusumoto K. Bone tissue engineering using human adipose-derived stem cells and honeycomb collagen scaffold. J Biomed Mater Res A. 2008;84:191–7.

    Google Scholar 

  6. Janicki P, Schmidmaier G. What should be the characteristics of the ideal bone graft substitute? Combining scaffolds with growth factors and/or stem cells. Injury. 2011;2:77–81.

    Article  Google Scholar 

  7. Hench LL, Paschall HA. Direct chemical bond of bioactive glass-ceramic materials to bone and muscle. J Biomed Mater Res A. 1973;3:25–42.

    Article  Google Scholar 

  8. Jarcho M, Kay JF, Gumaer KI, Doremus RH, Drobeck HP. Tissue, cellular and subcellular events at a bone-ceramic hydroxylapatite interface. J Bioeng. 1977;2:79–92.

    Google Scholar 

  9. Porter AE, Botelho CM, Lopes MA, Santos JD, Best SM, Bonfield W. Ultrastructural comparison of dissolution and apatite precipitation on hydroxyapatite and silicon-substituted hydroxyapatite in vitro and in vivo. J Biomed Mater Res A. 2004;69:670–9.

    Article  Google Scholar 

  10. Buehler J, Chappuis P, Saffar JL, Tsouderos Y, Vignery A. Strontium ranelate inhibits bone resorption while maintaining bone formation in alveolar bone in monkeys (Macaca fascicularis). Bone. 2001;1:176–9.

    Article  Google Scholar 

  11. Blake DGM, Zivanovic MA, McEwan AJ, Ackery DM. Sr-89 therapy: strontium kinetics in disseminated carcinoma of the prostate. Eur J Nucl Med. 1986;9:447–54.

    Google Scholar 

  12. Wong CT, Lu WW, Chan WK, Cheung KMC, Luk KDK, Lu DS, et al. In vivo cancellous bone remodeling on a strontium-containing hydroxyapatite (sr-HA) bioactive cement. J Biomed Mater Res A. 2004;68:513–21.

    Article  CAS  Google Scholar 

  13. Yang F, Yang D, Tu J, Zheng Q, Cai L, Wang L. Strontium enhances osteogenic differentiation of mesenchymal stem cells and in vivo bone formation by activating Wnt/catenin signaling. Stem Cells. 2011;6:981–91.

    Article  Google Scholar 

  14. Li Y, Li Q, Zhu S, Luo E, Li J, Feng G, et al. The effect of strontium-substituted hydroxyapatite coating on implant fixation in ovariectomized rats. Biomaterials. 2010;34:9006–14.

    Article  Google Scholar 

  15. Zreiqat H, Ramaswamy Y, Wu C, Paschalidis A, Lu Z, James B, et al. The incorporation of strontium and zinc into a calcium–silicon ceramic for bone tissue engineering. Biomaterials. 2010;12:3175–84.

    Article  Google Scholar 

  16. Gentleman E, Fredholm YC, Jell G, Lotfibakhshaiesh N, O’Donnell MD, Hill RG, et al. The effects of strontium-substituted bioactive glasses on osteoblasts and osteoclasts in vitro. Biomaterials. 2010;14:3949–56.

    Article  Google Scholar 

  17. Park J-W, Kim H-K, Kim Y-J, Jang J-H, Song H, Hanawa T. Osteoblast response and osseointegration of a Ti–6Al–4V alloy implant incorporating strontium. Acta Biomater. 2010;7:2843–51.

    Article  Google Scholar 

  18. Varma HK, Sivakumar R. Preparation and characterisation of free flowing hydroxyapatite powders. Phosphorous Res Bull. 1996;6:35–8.

    CAS  Google Scholar 

  19. Mohan BG, Shenoy SJ, Babu SS, Varma HK, John A. Strontium calcium phosphate for the repair of leporine (Oryctolagus cuniculus) ulna segmental defect. J Biomed Mater Res A. 2013;101:261–71.

    Google Scholar 

  20. Dorsey SM, Lin-Gibson S, Simon CG Jr. X-ray microcomputed tomography for the measurement of cell adhesion and proliferation in polymer scaffolds. Biomaterials. 2009;16:2967–74.

    Article  Google Scholar 

  21. Guilak F. Volume and surface area measurement of viable chondrocytes in situ using geometric modelling of serial confocal sections. J Microsc. 1994;173:245–56.

    Article  CAS  Google Scholar 

  22. Kokubo T, Kim H-M, Kawashita M. Novel bioactive materials with different mechanical properties. Biomaterials. 2003;13:2161–75.

    Article  Google Scholar 

  23. Dahl SG, Allain P, Marie PJ, Mauras Y, Boivin G, Ammann P, et al. Incorporation and distribution of strontium in bone. Bone. 2001;4:446–53.

    Article  Google Scholar 

  24. Canalis E, Hott M, Deloffre P, Tsouderos Y, Marie PJ. The divalent strontium salt S12911 enhances bone cell replication and bone formation in vitro. Bone. 1996;6:517–23.

    Article  Google Scholar 

  25. Likins RC, Posner AS, Kunde ML, Craven DL. Comparative metabolism of calcium and strontium in the rat. Arch Biochem Biophys. 1959;83:472–81.

    Article  CAS  Google Scholar 

  26. Hench LL, Wilson J. An introduction to bioactive ceramics. New york: World Scientific; 1993. p. 7.

    Book  Google Scholar 

  27. Tsuruga E, Takita H, Itoh H, Wakisaka Y, Kuboki Y. Pore size of porous hydroxyapatite as the cell-substratum controls BMP-induced osteogenesis. J Biochem. 1997;121:317–24.

    Article  CAS  Google Scholar 

  28. Dudas JR, Marra KG, Cooper GM, Penascino VM, Mooney MP, Jiang S, Rubin JP, et al. The osteogenic potential of adipose-derived stem cells for the repair of rabbit calvarial defects. Ann Plast Surg. 2006;56:543–8.

    Article  CAS  Google Scholar 

  29. Zangi L, Rivkin R, Kassis I, Levdansky L, Marx G, Gorodetsky R. High-yield isolation, expansion, and differentiation of rat bone marrow-derived mesenchymal stem cells with fibrin microbeads. Tissue Eng. 2006;8:2343–54.

    Article  Google Scholar 

  30. Mitchell JB, McIntosh K, Zvonic S, Garrett S, Floyd ZE, Kloster A, et al. Immunophenotype of human adipose-derived cells: temporal changes in stromal-associated and stem cell-associated markers. Stem Cells. 2006;2:376–85.

    Article  Google Scholar 

  31. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 2002;12:4279–95.

    Article  Google Scholar 

  32. Tan W, Sendemir-Urkmez A, Fahrner LJ, Jamison R, Leckband D, Boppart SA. Structural and functional optical imaging of three-dimensional engineered tissue development. Tissue Eng. 2004;12:1747–56.

    Article  Google Scholar 

  33. Ho ST, Hutmacher DW. A comparison of micro CT with other techniques used in the characterization of scaffolds. Biomaterials. 2006;8:1362–76.

    Article  Google Scholar 

  34. Valentijn AJ, Zouq N, Gilmore AP. Anoikis. Biochem Soc Trans. 2004;32:421–5.

    Article  CAS  Google Scholar 

  35. Marie PJ, Ammann P, Boivin G, Rey C. Mechanisms of action and therapeutic potential of strontium in bone. Calcif Tissue Int. 2001;69:121–9.

    Article  CAS  Google Scholar 

  36. Zhang R, Ma PX. Poly(alpha-hydroxyl acids)/hydroxyapatite porous composites for bone-tissue engineering. I. Preparation and morphology. J Biomed Mater Res. 1999;44:446–55.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors acknowledge The Director SCTIMST and Head, BMT Wing for the facilities provided; DBT for financial assistance; UGC for the Senior Research Fellowship; Mr. Rana V Raj for isolating adipose tissue; Dr. Anil Kumar TV for confocal analysis, Mr. Nishad KV for technical support, Mr. Renjith P Nair for FACS analysis and Dental products laboratory for Micro-CT facility.

Author information

Authors and Affiliations

  1. Transmission Electron Microscopy Laboratory, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences & Technology, Thiruvananthapuram, 695012, India

    Beena Gopalan Mohan, Sivadasan Suresh Babu, Hari Krishna Varma & Annie John

Authors
  1. Beena Gopalan Mohan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sivadasan Suresh Babu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hari Krishna Varma
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Annie John
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Annie John.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mohan, B.G., Suresh Babu, S., Varma, H.K. et al. In vitro evaluation of bioactive strontium-based ceramic with rabbit adipose-derived stem cells for bone tissue regeneration. J Mater Sci: Mater Med 24, 2831–2844 (2013). https://doi.org/10.1007/s10856-013-5018-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10856-013-5018-y

Keywords

Search

Navigation