Annals of Biomedical Engineering

, Volume 45, Issue 1, pp 210–223 | Cite as

Alginate Sulfate–Nanocellulose Bioinks for Cartilage Bioprinting Applications

  • Michael Müller
  • Ece Öztürk
  • Øystein Arlov
  • Paul Gatenholm
  • Marcy Zenobi-Wong
Additive Manufacturing of Biomaterials, Tissues, and Organs

Abstract

One of the challenges of bioprinting is to identify bioinks which support cell growth, tissue maturation, and ultimately the formation of functional grafts for use in regenerative medicine. The influence of this new biofabrication technology on biology of living cells, however, is still being evaluated. Recently we have identified a mitogenic hydrogel system based on alginate sulfate which potently supports chondrocyte phenotype, but is not printable due to its rheological properties (no yield point). To convert alginate sulfate to a printable bioink, it was combined with nanocellulose, which has been shown to possess very good printability. The alginate sulfate/nanocellulose ink showed good printing properties and the non-printed bioink material promoted cell spreading, proliferation, and collagen II synthesis by the encapsulated cells. When the bioink was printed, the biological performance of the cells was highly dependent on the nozzle geometry. Cell spreading properties were maintained with the lowest extrusion pressure and shear stress. However, extruding the alginate sulfate/nanocellulose bioink and chondrocytes significantly compromised cell proliferation, particularly when using small diameter nozzles and valves.

Keywords

Bioprinting Nanocellulose Alginate sulfate Cartilage tissue engineering 

Supplementary material

10439_2016_1704_MOESM1_ESM.docx (13 kb)
Supplementary material 1 (DOCX 13 kb)
10439_2016_1704_MOESM2_ESM.pdf (1.3 mb)
Supplementary material 2 (PDF 1286 kb)

References

  1. 1.
    Almeida, C. R., T. Serra, M. I. Oliveira, J. A. Planell, M. A. Barbosa, and M. Navarro. Impact of 3-D printed PLA- and chitosan-based scaffolds on human monocyte/macrophage responses: unraveling the effect of 3-D structures on inflammation. Acta Biomater. 10:613–622, 2014.CrossRefPubMedGoogle Scholar
  2. 2.
    Arlov, O., F. L. Aachmann, E. Feyzi, A. Sundan, and G. Skjak-Braek. The impact of chain length and flexibility in the interaction between sulfated alginates and HGF and FGF-2. Biomacromolecules 16:3417–3424, 2015.CrossRefPubMedGoogle Scholar
  3. 3.
    Arlov, O., F. L. Aachmann, A. Sundan, T. Espevik, and G. Skjak-Braek. Heparin-like properties of sulfated alginates with defined sequences and sulfation degrees. Biomacromolecules 15:2744–2750, 2014.CrossRefPubMedGoogle Scholar
  4. 4.
    Chang, R., J. Nam, and W. Sun. Direct cell writing of 3d microorgan for in vitro pharmacokinetic model. Tissue Eng. Part C 14:157–166, 2008.CrossRefGoogle Scholar
  5. 5.
    Chang, R., J. Nam, and W. Sun. Effects of dispensing pressure and nozzle diameter on cell survival from solid freeform fabrication-based direct cell writing. Tissue Eng. Part A 14:41–48, 2008.CrossRefPubMedGoogle Scholar
  6. 6.
    Chen, F. H., K. T. Rousche, and R. S. Tuan. Technology insight: adult stem cells in cartilage regeneration and tissue engineering. Nat. Clin. Pract. Rheumatol. 2:373–382, 2006.CrossRefPubMedGoogle Scholar
  7. 7.
    Freeman, I., A. Kedem, and S. Cohen. The effect of sulfation of alginate hydrogels on the specific binding and controlled release of heparin-binding proteins. Biomaterials 29:3260–3268, 2008.CrossRefPubMedGoogle Scholar
  8. 8.
    Jakob, M., O. Demarteau, D. Schafer, B. Hintermann, W. Dick, M. Heberer, and I. Martin. Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro. J. Cell. Biochem. 81:368–377, 2001.CrossRefPubMedGoogle Scholar
  9. 9.
    Jungst, T., W. Smolan, K. Schacht, T. Scheibel, and J. Groll. Strategies and molecular design criteria for 3d printable hydrogels. Chem. Rev. 116(3):1496–1539, 2015.CrossRefPubMedGoogle Scholar
  10. 10.
    Kesti, M., M. Muller, J. Becher, M. Schnabelrauch, M. D’Este, D. Eglin, and M. Zenobi-Wong. A versatile bioink for three-dimensional printing of cellular scaffolds based on thermally and photo-triggered tandem gelation. Acta Biomater. 11:162–172, 2015.CrossRefPubMedGoogle Scholar
  11. 11.
    Khalil, S., and W. Sun. Bioprinting endothelial cells with alginate for 3d tissue constructs. J. Biomech. Eng. 131:111002, 2009.CrossRefPubMedGoogle Scholar
  12. 12.
    Klein, T. J., S. C. Rizzi, K. Schrobback, J. C. Reichert, J. E. Jeon, R. W. Crawford, and D. W. Hutmacher. Long-term effects of hydrogel properties on human chondrocyte behavior. Soft Matter 6:5175–5183, 2010.CrossRefGoogle Scholar
  13. 13.
    Madry, H., A. Rey-Rico, J. K. Venkatesan, B. Johnstone, and M. Cucchiarini. Transforming growth factor beta-releasing scaffolds for cartilage tissue engineering. Tissue Eng. Part B 20:106–125, 2013.CrossRefGoogle Scholar
  14. 14.
    Malda, J., J. Visser, F. P. Melchels, T. Jungst, W. E. Hennink, W. J. A. Dhert, J. Groll, and D. W. Hutmacher. 25th anniversary article: engineering hydrogels for biofabrication. Adv. Mater. 25:5011–5028, 2013.CrossRefPubMedGoogle Scholar
  15. 15.
    Markstedt, K., A. Mantas, I. Tournier, and H. Martinez. Avila, D. Hagg and P. Gatenholm. 3d bioprinting human chondrocytes with nanocellulose–alginate bioink for cartilage tissue engineering applications. Biomacromolecules 16:1489–1496, 2015.CrossRefPubMedGoogle Scholar
  16. 16.
    Melchels, F. P. W., W. J. A. Dhert, D. W. Hutmacher, and J. Malda. Development and characterisation of a new bioink for additive tissue manufacturing. J. Mater. Chem. B 2:2282–2289, 2014.CrossRefGoogle Scholar
  17. 17.
    Mhanna, R., A. Kashyap, G. Palazzolo, Q. Vallmajo-Martin, J. Becher, S. Moller, M. Schnabelrauch, and M. Zenobi-Wong. Chondrocyte culture in three dimensional alginate sulfate hydrogels promotes proliferation while maintaining expression of chondrogenic markers. Tissue Eng. Part A 20:1454–1464, 2014.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Muller, M., J. Becher, M. Schnabelrauch, and M. Zenobi-Wong. Nanostructured pluronic hydrogels as bioinks for 3d bioprinting. Biofabrication 7:035006, 2015.CrossRefPubMedGoogle Scholar
  19. 19.
    Owczarczak, A. B., S. O. Shuford, S. T. Wood, S. Deitch, and D. Dean. Creating transient cell membrane pores using a standard inkjet printer. J. Vis. Expo 2012. doi:10.3791/3681.Google Scholar
  20. 20.
    Öztürk, E., Ø. Arlov, S. Aksel, L. Li, D. M. Ornitz, G. Skjåk-Bræk, and M. Zenobi-Wong. Sulfated hydrogel matrices direct mitogenicity and maintenance of chondrocyte phenotype through activation of fgf signaling. Adv. Funct. Mater. 26:3649–3662, 2016.CrossRefGoogle Scholar
  21. 21.
    Snyder, J., A. R. Son, Q. Hamid, C. Y. Wang, Y. G. Lui, and W. Sun. Mesenchymal stem cell printing and process regulated cell properties. Biofabrication 7(4):044106, 2015.CrossRefPubMedGoogle Scholar
  22. 22.
    Song, S. J., J. Choi, Y. D. Park, J. J. Lee, S. Y. Hong, and K. Sun. A three-dimensional bioprinting system for use with a hydrogel-based biomaterial and printing parameter characterization. Artif. Organs 34:1044–1048, 2010.CrossRefPubMedGoogle Scholar
  23. 23.
    Spiller, K. L., Y. Liu, J. L. Holloway, S. A. Maher, Y. Cao, W. Liu, G. Zhou, and A. M. Lowman. A novel method for the direct fabrication of growth factor-loaded microspheres within porous nondegradable hydrogels: controlled release for cartilage tissue engineering. J. Controlled Release 157:39–45, 2012.CrossRefGoogle Scholar
  24. 24.
    Wust, S., M. E. Godla, R. Muller, and S. Hofmann. Tunable hydrogel composite with two-step processing in combination with innovative hardware upgrade for cell-based three-dimensional bioprinting. Acta Biomater. 10:630–640, 2014.CrossRefPubMedGoogle Scholar
  25. 25.
    Wust, S., R. Muller, and S. Hofmann. Controlled positioning of cells in biomaterials-approaches towards 3d tissue printing. Journal of functional biomaterials 2:119–154, 2011.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Xu, T., J. Jin, C. Gregory, J. J. Hickman, and T. Boland. Inkjet printing of viable mammalian cells. Biomaterials 26:93–99, 2005.CrossRefPubMedGoogle Scholar
  27. 27.
    Zhao, Y., Y. Li, S. S. Mao, W. Sun, and R. Yao. The influence of printing parameters on cell survival rate and printability in microextrusion-based 3d cell printing technology. Biofabrication 7:045002, 2015.CrossRefPubMedGoogle Scholar

Copyright information

© Biomedical Engineering Society 2016

Authors and Affiliations

  1. 1.Cartilage Engineering + Regeneration Laboratory, Department of Health Sciences & TechnologyETH ZürichZurichSwitzerland
  2. 2.Department of BiotechnologyNorwegian University of Science and TechnologyTrondheimNorway
  3. 3.Department of Chemical and Biological Engineering, Wallenberg Wood Science Center and 3D Bioprinting CenterChalmers University of TechnologyGothenburgSweden

Personalised recommendations