Journal of Materials Science: Materials in Medicine

, Volume 23, Issue 11, pp 2821–2830 | Cite as

Peripheral mineralization of a 3D biodegradable tubular construct as a way to enhance guidance stabilization in spinal cord injury regeneration

  • A. L. OliveiraEmail author
  • E. C. Sousa
  • N. A. Silva
  • N. Sousa
  • A. J. Salgado
  • R. L. Reis


Spinal cord injuries (SCI) present a major challenge to therapeutic development due to its complexity. Combinatorial approaches using biodegradable polymers that can simultaneously provide a tissue scaffold, a cell vehicle, and a reservoir for sustained drug delivery have shown very promising results. In our previous studies we have developed a novel hybrid system consisting of starch/poly-e-caprolactone (SPCL) semi-rigid tubular porous structure, based on a rapid prototyping technology, filled by a gellan gum hydrogel concentric core for the regeneration within spinal-cord injury sites. In the present work we intend to promote enhanced osteointegration on these systems by pre-mineralizing specifically the external surfaces of the SPCL tubular structures, though a biomimetic strategy, using a sodium silicate gel as nucleating agent. The idea is to create two different cell environments to promote axonal regeneration in the interior of the constructs while inducing osteogenic activity on its external surface. By using a Teflon cylinder to isolate the interior of the scaffold, it was possible to observe the formation of a bone-like poorly crystalline carbonated apatite layer continuously formed only in the external side of the tubular structure. This biomimetic layer was able to support the adhesion of Bone Marrow Mesenchymal Stem Cells, which have gone under cytoskeleton reorganization in the first hours of culture when compared to cells cultured on uncoated scaffolds. This strategy can be a useful route for locally stimulate bone tissue regeneration and facilitating early bone ingrowth.


Apatite Simulated Body Fluid Simulated Body Fluid Solution Apatite Layer Spinal Cord Injury 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



Portuguese Foundation for Science and Technology under POCTI and/or FEDER programs (pre-doctoral fellowship to Nuno A. Silva, SFRH/BD/40684/2007, post-doctoral fellowship to Ana L. Oliveira, SFRH/BPD/39102/2007, and Ciência 2007 Program to António J. Salgado); Foundation Calouste de Gulbenkian to funds attributed to António J. Salgado under the scope of the Gulbenkian Programme to Support Research in the Life Sciences.


  1. 1.
    Profyris C, Cheema SS, Zang DW, Azari MF, Boyle K, Petratos S. Degenerative and regenerative mechanisms governing spinal cord injury. Neurobiol Dis. 2004;15:415–36.CrossRefGoogle Scholar
  2. 2.
    Hurlbert RJ. Methylprednisolone for acute spinal cord injury: an inappropriate standard of care. J Neurosurg. 2000;93:1–7.CrossRefGoogle Scholar
  3. 3.
    David S, Lacroix S. Molecular approaches to spinal cord repair. Annu Rev Neurosci. 2003;26:411–40.CrossRefGoogle Scholar
  4. 4.
    Jones LL, Oudega M, Bunge MB, Tuszynski MH. Neurotrophic factors, cellular bridges and gene therapy for spinal cord injury. J Physiol Lond. 2001;533:83–9.CrossRefGoogle Scholar
  5. 5.
    Zurita M, Vaquero J, Oya S, Montilla J. Functional recovery in chronic paraplegic rats after co-grafts of fetal brain and adult peripheral nerve tissue. Surg Neurol. 2001;55:249–54.CrossRefGoogle Scholar
  6. 6.
    Kakulas BA. Neuropathology: the foundation for new treatments in spinal cord injury. Spinal Cord. 2004;42:549–63.CrossRefGoogle Scholar
  7. 7.
    Myckatyn TM, Mackinnon SE, McDonald JW. Stem cell transplantation and other novel techniques for promoting recovery from spinal cord injury. Transpl Immunol. 2004;12:343–58.CrossRefGoogle Scholar
  8. 8.
    Silva NA, Salgado AJ, Sousa RA, Oliveira JT, Pedro AJ, Leite-Almeida H, et al. Development and characterization of a novel hybrid tissue engineering-based scaffold for spinal cord injury repair. Tissue Eng Part A. 2010;16:45–54.CrossRefGoogle Scholar
  9. 9.
    Woerly S, Doan VD, Sosa N, de Vellis J, Espinosa-Jeffrey A. Prevention of gliotic scar formation by NeuroGel (TM) allows partial endogenous repair of transected cat spinal cord. J Neurosci Res. 2004;75:262–72.CrossRefGoogle Scholar
  10. 10.
    Hejcl A, Lesny P, Pradny M, Michalek J, Jendelova P, Stulik J, et al. Biocompatible hydrogels in spinal cord injury repair. Physiol Res. 2008;57:S121–32.Google Scholar
  11. 11.
    Sykova E, Jendelova P, Urdzikova L, Lesny P, Hejcl A. Bone marrow stem cells and polymer hydrogels-two strategies for spinal cord injury repair. Cell Mol Neurobiol. 2006;26:1113–29.CrossRefGoogle Scholar
  12. 12.
    Macaya D, Spector M. Injectable hydrogel materials for spinal cord regeneration: a review. Biomed Mater. 2012;7:012001.CrossRefGoogle Scholar
  13. 13.
    Silva NA, Salgado AJ, Sousa RA, Oliveira JT, Neves NM, Mano JF, et al. Starch/gellan gum hybrid 3D guidance systems for spinal cord injury regeneration: Scaffolds processing, characterization and biological evaluation. Tissue Eng Part A. 2008;14:779.Google Scholar
  14. 14.
    Silva NA, Sousa RA, Oliveira JT, Fraga JS, Fontes M, Cerqueira R, et al. Benefits of Spine Stabilization with Biodegradable Scaffolds in Spinal Cord Injured Rats. Tissue Engineering Part C. 2012. doi: 10.1089/ten.TEC.2012.0264.
  15. 15.
    Silva NA, Sousa RA, Pires AO, Sousa N, Salgado AJ, Reis RL. Interactions between Schwann and olfactory ensheathing cells with a starch/polycaprolactone scaffold aimed at spinal cord injury repair. J Biomed Mater Res A. 2012;100A:470–6.CrossRefGoogle Scholar
  16. 16.
    Rey C. Calcium phosphate biomaterials bone mineral. Differences in composition structures and properties. Biomaterials. 1990;11:13.CrossRefGoogle Scholar
  17. 17.
    Oliveira AL, Mano JF, Reis RL. Nature-inspired calcium phosphate coatings: present status and novel advances in the science of mimicry. Curr Opin Solid State Mater. 2003;7:309–18.CrossRefGoogle Scholar
  18. 18.
    Abe Y, Kokubo T, Yamamuro T. Apatite coating on ceramics, metals and polymers utilizing a biological process. J Mater Sci Mater Med. 1990;1:233–8.CrossRefGoogle Scholar
  19. 19.
    Reis RL, Cunha AM, Allan PS, Bevis MJ. Mechanical behaviour of injection-moulded starch based polymers. Polym Adv Technol. 1996;7:784–90.CrossRefGoogle Scholar
  20. 20.
    Reis RL, Cunha AM. Characterization of two biodegradable polymers of potential application within the biomaterials field. J Mater Sci Mater Med. 1995;6:786–92.CrossRefGoogle Scholar
  21. 21.
    Oliveira AL, Elvira C, Vásquez B, San Roman J, Reis RL. Surface modifications tailors the characteristics of biomimetic coatings nucleated on starch based polymers. J Mater Sci Mater Med. 1999;10:827.CrossRefGoogle Scholar
  22. 22.
    Elvira C, Mano JF, San Roman J, Reis RL. Starch-based biodegradable hydrogels with potential biomedical applications as drug delivery systems. Biomaterials. 2002;23:1955–66.CrossRefGoogle Scholar
  23. 23.
    Gomes ME, Holtorf HL, Reis RL, Mikos AG. Influence of the porosity of starch-based fiber mesh scaffolds on the proliferation and osteogenic differentiation of bone marrow stromal cells cultured in a flow perfusion bioreactor. Tissue Eng. 2006;12:801–9.CrossRefGoogle Scholar
  24. 24.
    Gomes ME, Sikavitsas VI, Behravesh E, Reis RL, Mikos Antonios G. Effect of flow perfusion on the osteogenic differentiation of bone marrow stromal cells cultured on starch based three-dimensional scaffolds. J Biomed Mater Res A. 2003;67:87–95.CrossRefGoogle Scholar
  25. 25.
    Tuzlakoglu K, Bolgen N, Salgado AJ, Gomes ME, Piskin E, Reis RL. Nano- and micro-fiber combined scaffolds: a new architecture for bone tissue engineering. J Mater Sci Mater Med. 2005;16:1099–104.CrossRefGoogle Scholar
  26. 26.
    Santos MI, Fuchs S, Gomes ME, Unger RE, Reis RL, Kirkpatrick CJ. Response of micro- and macrovascular endothelial cells to starch-based fiber meshes for bone tissue engineering. Biomaterials. 2007;28:240–8.CrossRefGoogle Scholar
  27. 27.
    Silva N, Salgado AJ, Sousa RA, Oliveira JO, Pedro AJ, Mastronardi F, et al. Development and characterization of a novel hybrid tissue engineering based scaffold for spinal cord injury repair. Tissue Eng. 2008;16:45–54.Google Scholar
  28. 28.
    Oliveira AL, Malafaya PB, Reis RL. Sodium silicate gel as a precursor for the in vitro nucleation and growth of a bone-like apatite coating in compact and porous polymeric structures. Biomaterials. 2003;24:2575–84.CrossRefGoogle Scholar
  29. 29.
    Kokubo T, Takadama H. How useful is SBF in predicting in vivo bone bioactivity? Biomaterials. 2006;27:2907–15.CrossRefGoogle Scholar
  30. 30.
    Kokubo T, Kushitani H, Sakka S, Kitsugi T, Yamamuro T. Solutions able to reproduce in vivo surface-structure changes in bioactive glass-ceramic A-W. J Biomed Mater Res. 1990;24:721–34.CrossRefGoogle Scholar
  31. 31.
    Oliveira JT, Gardel LS, Rada T, Martins L, Gomes ME, Reis RL. Injectable gellan gum hydrogels with autologous cells for the treatment of rabbit articular cartilage defects. J Orthop Res. 2010;28:1193–9.CrossRefGoogle Scholar
  32. 32.
    Gomes ME, Sikavitsas VI, Behravesh E, Reis RL, Mikos AG. Effect of flow perfusion on the osteogenic differentiation of bone marrow stromal cells cultured on starch-based three-dimensional scaffolds. J Biomed Mater Res A. 2003;67A:87–95.CrossRefGoogle Scholar
  33. 33.
    Mendes SC, Bezemer J, Claase MB, Grijpma DW, Bellia G, Degli-Innocenti F, et al. Evaluation of two biodegradable polymeric systems as substrates for bone tissue engineering. Tissue Eng. 2003;9:S91–101.CrossRefGoogle Scholar
  34. 34.
    Oliveira AL, Costa SA, Sousa RA, Reis RL. Nucleation and growth of biomimetic apatite layers on 3D plotted biodegradable polymeric scaffolds: effect of static and dynamic coating conditions. Acta Biomater. 2009;5:1626–38.CrossRefGoogle Scholar
  35. 35.
    Boskey AL, Posner AS. Magnesium stabilization of amorphous calcium-phosphate: a kinetic study. Mater Res Bull. 1974;9:907–16.CrossRefGoogle Scholar
  36. 36.
    Takadama H, Kim HM, Kokubo T, Nakamura T. Mechanism of biomineralization of apatite on a sodium silicate glass: TEM-EDX study in vitro. Chem Mater. 2001;13:1108–13.CrossRefGoogle Scholar
  37. 37.
    Oliveira AL, Reis RL. Pre-mineralisation of starch/polycrapolactone bone tissue engineering scaffolds by a calcium-silicate-based process. J Mater Sci Mater Med. 2004;15:533–40.CrossRefGoogle Scholar
  38. 38.
    Vallet-Regi M, Romero AM, Ragel CV, LeGeros RZ. XRD, SEM-EDS, and FTIR studies of in vitro growth of an apatite-like layer on sol-gel glasses. J Biomed Mater Res. 1999;44:416–21.CrossRefGoogle Scholar
  39. 39.
    Rehman I, Bonfield W. Characterization of hydroxyapatite and carbonated apatite by photo acoustic FTIR spectroscopy. J Mater Sci Mater Med. 1997;8:1–4.CrossRefGoogle Scholar
  40. 40.
    Oliveira AL, Malafaya PB, Costa SA, Sousa RA, Reis RL. Micro-computed tomography (micro-CT) as a potential tool to assess the effect of dynamic coating routes on the formation of biomimetic apatite layers on 3D-plotted biodegradable polymeric scaffolds. J Mater Sci Mater Med. 2007;18:211–23.CrossRefGoogle Scholar
  41. 41.
    Hirota M, Hayakawa T, Ametani A, Kuboki Y, Sato M, Tohnai I. The effect of hydroxyapatite-coated titanium fiber web on human osteoblast functional activity. Int J Oral Maxillofac Implants. 2011;26:245–50.Google Scholar
  42. 42.
    Salgado AJ, Figueiredo JE, Coutinho OP, Reis RL. Biological response to pre-mineralized starch based scaffolds for bone tissue engineering. J Mater Sci Mater Med. 2005;16:267–75.CrossRefGoogle Scholar
  43. 43.
    Ong JL, Villarreal DR, Cavin R, Ma K. Osteoblast responses to as-deposited and heat treated sputtered CaP surfaces. J Mater Sci Mater Med. 2001;12:491–5.CrossRefGoogle Scholar
  44. 44.
    ter Brugge PJ, Jansen JA. Initial interaction of rat bone marrow cells with non-coated and calcium phosphate coated titanium substrates. Biomaterials. 2002;23:3269–77.CrossRefGoogle Scholar
  45. 45.
    Ramires PA, Giuffrida A, Milella E. Three-dimensional reconstruction of confocal laser microscopy images to study the behaviour of osteoblastic cells grown on biomaterials. Biomaterials. 2002;23:397–406.CrossRefGoogle Scholar
  46. 46.
    Araujo JV, Martins A, Leonor IB, Pinho ED, Reis RL, Neves NM. Surface controlled biomimetic coating of polycaprolactone nanofiber meshes to be used as bone extracellular matrix analogues. J Biomat Sci Polym Ed. 2008;19:1261–78.CrossRefGoogle Scholar
  47. 47.
    Oliveira AL, Alves CM, Reis RL. Cell adhesion and proliferation on biomimetic calcium-phosphate coatings produced by a sodium silicate gel methodology. J Mater Sci Mater Med. 2002;13:1181–8.CrossRefGoogle Scholar
  48. 48.
    Barrere F, van der Valk CM, Dalmeijer RAJ, van Blitterswijk CA, de Groot K, Layrolle P. In vitro and in vivo degradation of biomimetic octacalcium phosphate and carbonate apatite coatings on titanium implants. J Biomed Mater Res A. 2003;64A:378–87.CrossRefGoogle Scholar
  49. 49.
    Hayakawa T, Takahashi K, Yoshinari M, Okada H, Yamamoto H, Sato M, et al. Trabecular bone response to titanium implants with a thin carbonate-containing apatite coating applied using the molecular precursor method. Int J Oral Maxillofac Implants. 2006;21:851–8.Google Scholar
  50. 50.
    Vasudev DV, Ricci JL, Sabatino C, Li PJ, Parsons JR. In vivo evaluation of a biomimetic apatite coating grown on titanium surfaces. J Biomed Mater Res A. 2004;69A:629–36.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • A. L. Oliveira
    • 1
    • 2
    • 4
    Email author
  • E. C. Sousa
    • 1
    • 2
  • N. A. Silva
    • 1
    • 2
    • 3
  • N. Sousa
    • 2
    • 3
  • A. J. Salgado
    • 2
    • 3
  • R. L. Reis
    • 1
    • 2
  1. 1.3B’s Research Group—Biomaterials, Biodegradables and BiomimeticsUniversity of MinhoTaipas, GuimarãesPortugal
  2. 2.ICVS/3B’s—PT Government Associate LaboratoryBraga/GuimarãesPortugal
  3. 3.Life and Health Sciences Research Institute (ICVS), School of Health SciencesUniversity of MinhoBragaPortugal
  4. 4.Department of Health SciencesPortuguese Catholic UniversityViseuPortugal

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