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Heparinized hydroxyapatite/collagen three-dimensional scaffolds for tissue engineering

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Abstract

Currently, in bone tissue engineering research, the development of appropriate biomaterials for the regeneration of bony tissues is a major concern. Bone tissue is composed of a structural protein, collagen type I, on which calcium phosphate crystals are enclosed. For tissue engineering, one of the most applied strategies consists on the development and application of three dimensional porous scaffolds with similar composition to the bone. In this way, they can provide a physical support for cell attachment, proliferation, nutrient transport and new bone tissue infiltration. Hydroxyapatite is a calcium phosphate with a similar composition of bone and widely applied in several medical/dentistry fields. Therefore, in this study, hydroxyapatite three dimensional porous scaffolds were produced using the polymer replication method. Next, the porous scaffolds were homogeneously coated with a film of collagen type I by applying vacuum force. Yet, due to collagen degradability properties, it was necessary to perform an adequate crosslinking method. As a result, N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) was employed as an efficient and non-toxic crosslinking method in this research. The composites were characterized by means of SEM, DSC and TNBS. Furthermore, heparin was incorporated in order to accomplish sustained delivery of a growth factor of interest namely, bone morphogenetic proteins (BMP-2). BMP-2 binding and release of non-heparinized and heparinized scaffolds was evaluated at specific time points. The incorporation of heparin leads to a reduced initial burst phase when compared to the non heparinized materials. The results show a beneficial effect with the incorporation of heparin and its potential as a localized drug delivery system for the sustained release of growth factors.

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References

  1. Rose FR, Cyster LA, Grant DM, Scotchford CA, Howdle SM, Shakesheff KM. In vitro assessment of cell penetration into porous hydroxyapatite scaffolds with a central aligned channel. Biomaterials. 2004;25(24):5507–14.

    Article  CAS  PubMed  Google Scholar 

  2. Hong Z, Zhang P, He C, Qiu X, Liu A, Chen L, et al. Nano-composite of poly(l-lactide) and surface grafted hydroxyapatite: mechanical properties and biocompatibility. Biomaterials. 2005;26(32):6296–304.

    Article  CAS  PubMed  Google Scholar 

  3. Ignjatovic N, Tomic S, Dakic M, Miljkovic M, Plavsic M, Uskokovic D. Synthesis and properties of hydroxyapatite/poly-l-lactide composite biomaterials. Biomaterials. 1999;20(9):809–16.

    Article  CAS  PubMed  Google Scholar 

  4. Kothapalli CR, Shaw MT, Wei M. Biodegradable HA-PLA 3-D porous scaffolds: effect of nano-sized filler content on scaffold properties. Acta Biomater. 2005;1(6):653–62.

    Article  PubMed  Google Scholar 

  5. Liao S, Wang W, Uo M, Ohkawa S, Akasaka T, Tamura K, et al. A three-layered nano-carbonated hydroxyapatite/collagen/PLGA composite membrane for guided tissue regeneration. Biomaterials. 2005;26(36):7564–71.

    Article  CAS  PubMed  Google Scholar 

  6. Maeda H, Kasuga T, Hench LL. Preparation of poly(l-lactic acid)-polysiloxane-calcium carbonate hybrid membranes for guided bone regeneration. Biomaterials. 2006;27(8):1216–22.

    Article  CAS  PubMed  Google Scholar 

  7. Kong L, Gao Y, Cao W, Gong Y, Zhao N, Zhang X. Preparation and characterization of nano-hydroxyapatite/chitosan composite scaffolds. J Biomed Mater Res. 2005;75(2):275–82.

    Article  Google Scholar 

  8. Li XM, Feng QL, Cui FZ. In vitro degradation of porous nano-hydroxyapatite/collagen/PLLA scaffold reinforced by chitin fibres. Mater Sci Eng C-Biomim Supramol Syst. 2006;26(4):716–20.

    CAS  Google Scholar 

  9. Sachlos E, Gotora D, Czernuszka JT. Collagen scaffolds reinforced with biomimetic composite nano-sized carbonate-substituted hydroxyapatite crystals and shaped by rapid prototyping to contain internal microchannels. Tissue Eng. 2006;12(9):2479–87.

    Article  CAS  PubMed  Google Scholar 

  10. Zhu X, Eibl O, Scheideler L, Geis-Gerstorfer J. Characterization of nano hydroxyapatite/collagen surfaces and cellular behaviors. J Biomed Mater Res. 2006;79(1):114–27.

    Article  Google Scholar 

  11. Pieper JS, Hafmans T, Veerkamp JH, van Kuppevelt TH. Development of tailor-made collagen-glycosaminoglycan matrices: EDC/NHS crosslinking, and ultrastructural aspects. Biomaterials. 2000;21(6):581–93.

    Article  CAS  PubMed  Google Scholar 

  12. Pieper JS, Oosterhof A, Dijkstra PJ, Veerkamp JH, van Kuppevelt TH. Preparation and characterization of porous crosslinked collagenous matrices containing bioavailable chondroitin sulphate. Biomaterials. 1999;20(9):847–58.

    Article  CAS  PubMed  Google Scholar 

  13. Pieper JS, van Wachem PB, van Luyn MJA, Brouwer LA, Hafmans T, Veerkamp JH, et al. Attachment of glycosaminoglycans to collagenous matrices modulates the tissue response in rats. Biomaterials. 2000;21(16):1689–99.

    Article  CAS  PubMed  Google Scholar 

  14. Pieper JS, Hafmans T, van Wachem PB, van Luyn MJA, Brouwer LA, Veerkamp JH, et al. Loading of collagen-heparan sulfate matrices with bFGF promotes angiogenesis and tissue generation in rats. J Biomed Mater Res. 2002;62(2):185–94.

    Article  CAS  PubMed  Google Scholar 

  15. Wissink MJB, Beernink R, Pieper JS, Poot AA, Engbers GHM, Beugeling T, et al. Immobilization of heparin to EDC/NHS-crosslinked collagen. Characterization and in vitro evaluation. Biomaterials. 2001;22(2):151–63.

    Article  CAS  PubMed  Google Scholar 

  16. Wissink MJB, Beernink R, Poot AA, Engbers GHM, Beugeling T, van Aken WG, et al. Improved endothelialization of vascular grafts by local release of growth factor from heparinized collagen matrices. J Control Release. 2000;64(1–3):103–14.

    Article  CAS  PubMed  Google Scholar 

  17. Wissink MJB, Beernink R, Scharenborg NM, Poot AA, Engbers GHM, Beugeling T, et al. Endothelial cell seeding of (heparinized) collagen matrices: effects of bFGF pre-loading on proliferation (after low density seeding) and pro-coagulant factors. J Control Release. 2000;67(2–3):141–55.

    Article  CAS  PubMed  Google Scholar 

  18. Steffens GCM, Yao C, Prevel P, Markowicz M, Schenck P, Noah EM, et al. Modulation of angiogenic potential of collagen matrices by covalent incorporation of heparin and loading with vascular endothelial growth factor.Tissue Eng. 2004:1502–9.

  19. Yao C, Markowicz M, Pallua N, Magnus Noah E, Steffens G. The effect of cross-linking of collagen matrices on their angiogenic capability. Biomaterials. 2008;29(1):66–74.

    Article  CAS  PubMed  Google Scholar 

  20. Jiao YY, Ubrich N, Hoffart V, Marchand-Arvier M, Vigneron C, Hoffman M, et al. Preparation and characterization of heparin-loaded polymeric microparticles. Drug Dev Ind Pharm. 2002;28(8):1033–41.

    Article  CAS  PubMed  Google Scholar 

  21. Jiao YY, Ubrich N, Marchand-Arvier M, Vigneron C, Hoffman M, Lecompte T, et al. In vitro and in vivo evaluation of oral heparin-loaded polymeric nanoparticles in rabbits. Circulation. 2002;105(2):230–5.

    Article  CAS  PubMed  Google Scholar 

  22. Lee KW, Yoon JJ, Lee JH, Kim SY, Jung HJ, Kim SJ, et al. Sustained release of vascular endothelial growth factor from calcium-induced alginate hydrogels reinforced by heparin and chitosan. Transpl Proc. 2004;36(8):2464–5.

    Article  CAS  Google Scholar 

  23. SchroederTefft JA, Bentz H, Estridge TD. Collagen and heparin matrices for growth factor delivery. J Control Release. 1997;48(1):29–33.

    Article  CAS  Google Scholar 

  24. Teixeira S, Ferraz MP, Monteiro FJ. Biocompatibility of highly macroporous ceramic scaffolds: cell adhesion and morphology studies. J Mater Sci. 2008;19(2):855–9.

    CAS  Google Scholar 

  25. Edlund U, Dånmark S, Albertsson AC. A strategy for the covalent functionalization of resorbable polymers with heparin and osteoinductive growth factor. Biomacromolecules. 2008;9(3):901–5.

    Article  CAS  PubMed  Google Scholar 

  26. Teixeira S, Oliveira S, Ferraz M, Monteiro FJ. Three dimensional macroporous calcium phosphates scaffolds for bone tissue engineering. Key Eng Mater Trans Tech. 2008:947–50.

  27. Yamamoto M, Tabata Y, Hong L, Miyamoto S, Hashimoto N, Ikada Y. Bone regeneration by transforming growth factor beta1 released from a biodegradable hydrogel. J Control Release. 2000;64:133–42.

    Article  CAS  PubMed  Google Scholar 

  28. Teixeira S, Rodriguez MA, Pena P, De Aza AH, De Aza S, Ferraz MP, Monteiro FJ. Physical characterization of hydroxyapatite porous scaffolds for tissue engineering. Mater Sci Eng C. 2009;29(5):1510–4.

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by the Portuguese Foundation for Science and Technology (FCT) PhD grant SFRH/BD/17139/2004 and project FCT - POCTI/SAU - BMA/56061/2004. The authors would also like to thank Jun Liu for the assistance in the release experiments.

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Authors and Affiliations

  1. Divisão de Biomateriais, INEB—Instituto de Engenharia Biomédica, Rua do Campo Alegre 823, 4150-180, Porto, Portugal

    S. Teixeira, M. P. Ferraz & F. J. Monteiro

  2. Departamento de Engenharia Metalúrgica e Materiais, Faculdade de Engenharia, Universidade do Porto, Porto, Portugal

    S. Teixeira & F. J. Monteiro

  3. MIRA Institute for Biomedical Technology and Technical Medicine, Department of Polymer Chemistry and Biomaterials, University of Twente, P.O. Box 217, 7500AE, Enschede, The Netherlands

    L. Yang & P. J. Dijkstra

  4. Faculdade de Ciências da Saúde da Universidade Fernando Pessoa, Rua Carlos da Maia, 296, 4200-150, Porto, Portugal

    M. P. Ferraz

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  1. S. Teixeira
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  2. L. Yang
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  3. P. J. Dijkstra
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  4. M. P. Ferraz
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  5. F. J. Monteiro
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Correspondence to S. Teixeira.

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Teixeira, S., Yang, L., Dijkstra, P.J. et al. Heparinized hydroxyapatite/collagen three-dimensional scaffolds for tissue engineering. J Mater Sci: Mater Med 21, 2385–2392 (2010). https://doi.org/10.1007/s10856-010-4097-2

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  • DOI: https://doi.org/10.1007/s10856-010-4097-2

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