Skip to main content
SpringerLink
Log in

Cell response to collagen-calcium phosphate cement scaffolds investigated for nonviral gene delivery

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

Abstract

Collagen-hydroxyapatite (HA) scaffolds for the non-viral delivery of a plasmid encoding the osteoinductive protein bone morphogenetic protein (BMP)-7 were developed. The collagen-HA was obtained by the combination of calcium phosphate cement in a collagen template. The effect on cell behavior of increasing amounts of HA in the scaffolds was evaluated. Collagen-HA scaffolds containing 13, 23 or 83 wt% HA were prepared. Cell proliferation was reduced in the 83% HA scaffold after 1 day compared to 13 and 23% HA, but by 14 days the number of cells in 83% HA considerably increased. Alkaline phosphatase (ALP) activity was 8 times higher for the 83% HA scaffolds. BMP-7 plasmid was incorporated into the 83% HA scaffold. The transfection was low, although significant levels of BMP7 were expressed, associated with an increase in cell proliferation.

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

Similar content being viewed by others

References

  1. Cancedda R, Giannoni P, Mastrogiacomo M. A tissue engineering approach to bone repair in large animal models and in clinical practice. Biomaterials. 2007;28:4240–50.

    Article  CAS  Google Scholar 

  2. Stone KR, Steadman JR, Rodkey WG, Li S. Regeneration of meniscal cartilage with use of a collagen scaffold analysis of preliminary data. J Bone Joint Surg Am. 1997;79:17770–7.

    Google Scholar 

  3. Ahn S, Yoon H, Kim Y, Lee S, Chun W. Designed three dimensional collagen scaffolds for skin tissue regeneration. Tissue Eng C. 2010;16:813–20.

    Article  CAS  Google Scholar 

  4. Yu K, Takashi I, Yasuhiko T. Adipose tissue formation in collagen scaffolds with different biodegradabilities. J Biomater Sci Polym Ed. 2010;21:463–76.

    Article  Google Scholar 

  5. Sumita Y, Honda MJ, Ohara T, Tsuchiya S, Sagara H, Kagami H, Ueda M. Performance of collagen sponge as a 3-D scaffold for tooth tissue engineering. Biomaterials. 2006;27:3238–48.

    Article  CAS  Google Scholar 

  6. Madaghiele M, Sannino A, Yannas IV, Spector M. Collagen based matrices with axially oriented pores. J Biomed Mater Res. 2008;85A:757–67.

    Article  CAS  Google Scholar 

  7. O’Brien FJ, Harley BA, Yannas IV, Gibson L. Influence of freezing ratio on pore structure in freeze-dried collagen-GAG scaffolds. Biomaterials. 2004;25:1077–86.

    Article  Google Scholar 

  8. Wahl DA, Sachlo E, Liu C, Czernuszka JT. Controlling the processing of collagen-hydroxyapatite scaffolds for bone tissue engineering. J Mater Sci Mater Med. 2007;18:201–9.

    Article  CAS  Google Scholar 

  9. Yamauchi K, Goda T, Takuchi N, Einaga H, Tanabe T. Preparation of collagen/calcium phosphate multilayer sheet using enzymatic mineralization. Biomaterials. 2004;25:5481–9.

    Article  CAS  Google Scholar 

  10. Takahashi Y, Yamamoto M, Tabata Y. Enhanced osteoinduction by controlled release of bone morphogenetic protein 2 from biodegradable sponge compose of gelatin and β-tricalcium phosphate. Biomaterials. 2005;26:4856–65.

    Article  CAS  Google Scholar 

  11. Wozney JM, Rosen V. Bone morphogenetic protein and bone morphogenetic protein gene family in bone formation and repair. Clin Orthop Res. 1998;346:26–37.

    Article  Google Scholar 

  12. Azzam T, Domb AJ. Current developments in gene transfection agents. Curr Drug Del. 2004;1:165–93.

    Article  CAS  Google Scholar 

  13. Kofron MD, Laurencin CT. Bone tissue engineering by gene delivery. Adv Drug Del Rev. 2006;58:555–76.

    Article  CAS  Google Scholar 

  14. Maitra A. Calcium phosphate nanoparticles: second-generation nonviral vectors in gene therapy. Exp Rev Mol Diagn. 2005;5:893–905.

    Article  CAS  Google Scholar 

  15. Weadock KS, Miller EJ, Bellincampi LD, Zawadsky JP, Dunn MG. Physical crosslinking of collagen fibers: comparison of ultraviolet irradiation and dehydrothermal treatment. J Biomed Mater Res. 1995;29:1373–9.

    Article  CAS  Google Scholar 

  16. Yannas IV. Collagen and gelatin in the solid state. J Macromol Sci Revs Macromol Chem. 1972;C7:49–104.

    Google Scholar 

  17. Masson P. Some histological methods Trichrome stainings and their preliminary technique. J Tech Meth. 1929;12:75–90.

    Google Scholar 

  18. Flint MH, Lyons MF, Meaney MF, Williams DE. The Masson staining of collagen—an explanation of an apparent paradox. Histochem J. 1975;7:529–46.

    Article  CAS  Google Scholar 

  19. Haugh GH, Jaasma MJ, O’Brien FJ. The effect of dehydrothermal treatment on the mechanical and structural properties of collagen-GAG scaffolds. J Biomed Mater Res A. 2009;89:363–9.

    Google Scholar 

  20. Ginebra MP, Fernandez E, De Maeyer EAP, Verbeeck RMH, Boltong MG, Ginebra J, Driessens FCM, Planell JA. Setting reaction and hardening of an apatitic calcium phosphate cement. J Dent Res. 1997;76:905–12.

    Article  CAS  Google Scholar 

  21. Jones GL, Walton R, Czernuszka J, Griffiths SL, El Haj AJ, Cartmell SH. Primary human osteoblast culture on 3D porous collagen-hydroxyapatite scaffolds. J Biomed Mater Res A. 2010. doi:10.1002/jbm.a.32805.

  22. Engel E, del Valle S, Aparicio C, Altankov G, Asin L, Planell JA, Ginebra MP. Discerning the role of topography and ion exchange in cell response of bioactive tissue engineering scaffolds. Tissue Eng. 2008;14:1341–51.

    Article  CAS  Google Scholar 

  23. Laflamme C, Rouabhia M. Effect of BMP-2 and BMP-7 homodimers and a mixture of BMP-2/BMP-7 homodimers on oasteoblast adhesion and growth following culture on a collagen scaffold. Biomed Mater. 2008;3:1–10.

    Article  Google Scholar 

  24. Tsiridis E, Bhalla A, Ali Z, Gurav N, Heliotis M, Deb S, DiSilvio L. Enhancing the osteoinductive properties of hydroxyapatite by the addition of human mesenchymal stem cells, and recombinant human osteogenic protein-1 (BMP-7) in vitro. Injury. 2006;37:S25–32.

    Article  Google Scholar 

  25. Bright C, Park YS, Sieber AN, Kostuik JP, Leong KW. In vivo evaluation of plasmid DNA encoding OP-1 protein for spine fusion. Spine. 2006;31:2163–72.

    Article  Google Scholar 

  26. Zhou H, Wu T, Dong X, Wang Q, Shen J. Adsorption mechanism of BMP-7 on hydroxyapatite (001) surfaces. Biochem Biophys Res Commun. 2007;361:91–6.

    Article  CAS  Google Scholar 

  27. Boix T, Gómez-Morales J, Torrent-Burgués J, Monfort A, Puigdomènech P, Rodriguez-Celemente R. Adsorption of recombinant human bone morphogenetic protein rhBMP-2m onto hydroxyapatite. J Inorg Biochem. 2005;99:1043–50.

    Article  CAS  Google Scholar 

  28. Sokolova VV, Radtke I, Heumann R, Epple M. Effective transfection of cells with multi-shell calcium phosphate-DNA nanoparticles. Biomaterials. 2006;27:3147–53.

    Article  CAS  Google Scholar 

  29. Bish S, Bhakta G, Mitra S, Maitra A. pDNA loaded calcium phosphate nanoparticles: highly efficient non-viral vector for gene delivery. Int J Pharm. 2005;288:157–68.

    Article  Google Scholar 

  30. Roy I, Mitra S, Mitra A, Mozumdar S. Calcium phosphate nanoparticles as novel non-viral vectors for targeted gene delivery. Int J Pharm. 2003;250:25–33.

    Article  CAS  Google Scholar 

  31. He Q, Mitchell AR, Johnson SL, Wagner-Bartak C, Morcol T, Bell SJD. Calcium phosphate nanoparticle adjuvant. Clin Diag Lab Immunol. 2000;7:899–903.

    CAS  Google Scholar 

  32. Orrantia E, Chang PL. Intracellular distribution of DNA internalized through calcium phosphate precipitation. Exp Cell Res. 1990;190:170–4.

    Article  CAS  Google Scholar 

  33. Olton D, Li J, Wilson ME, Rogers T, Close J, Huang L, Kumta PN, Sfeir C. Nanostructured calcium phosphates (NanoCaPs) for non-viral gene delivery: influence of the synthesis parameters on transfection efficiency. Biomaterials. 2007;28:1267–79.

    Article  CAS  Google Scholar 

  34. Krebs MD, Salter E, Chen E, Kathleen AS, Alsberg E. Calcium phosphate-DNA nanoparticle gene delivery from alginate hydrogels induces in vivo osteogenesis. J Biomed Mater Res A. 2010;92:1131–8.

    Google Scholar 

  35. Keeney M, van den Beucken J, van der Kraan PM, Jansen JA, Pandit A. The ability of a collagen/calcium phosphate scaffold to act as its own vector for gene delivery and to promote bone formation via transfection with VEGF165. Biomaterials. 2010;31:2893–902.

    Article  CAS  Google Scholar 

  36. Luo D, Saltzman WM. Synthetic delivery systems. Nat Biotechnol. 2000;18:33–7.

    Article  CAS  Google Scholar 

  37. Wiethoff CM, Middaugh CR. Barriers to nonviral gene delivery. J Pharm Sci. 2003;92:203–17.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Spanish Ministry of Science and Education through project MAT2006-11516 and by the U.S. Department of Veterans Affairs, Veterans Health Administration, Rehabilitation Research and Development Service. The technical assistance of Alix Weaver greatly appreciated. The authors are grateful to Stryker Biotech, Hopkinton, MA for supplying the BMP-7 plasmid.

Author information

Authors and Affiliations

  1. Biomaterials, Biomechanics and Tissue Engineering Group, Department of Materials Science and Metallurgy, Technical University of Catalonia (UPC), 647 Diagonal Av, 08028, Barcelona, Spain

    R. A. Perez & M. P. Ginebra

  2. Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), María de Luna 11, Ed. CEEI, 50118, Zaragoza, Spain

    R. A. Perez & M. P. Ginebra

  3. Tissue Engineering, VA Boston Healthcare System, 150 S. Huntignton Av, Boston, MA, 02130, USA

    R. A. Perez & M. Spector

  4. Orthopedic Research Laboratory, Brigham and Women′s Hospital, Harvard Medical School, Boston, MA, 02115, USA

    M. Spector

Authors
  1. R. A. Perez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. P. Ginebra
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Spector
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. Spector.

Additional information

Paper selected for publication from the 23rd European Conference on Biomaterials, Tampere, Finland, September 2010.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Perez, R.A., Ginebra, M.P. & Spector, M. Cell response to collagen-calcium phosphate cement scaffolds investigated for nonviral gene delivery. J Mater Sci: Mater Med 22, 887–897 (2011). https://doi.org/10.1007/s10856-011-4308-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10856-011-4308-5

Keywords

Search

Navigation