Nanocomposite Clay-Based Bioinks for Skeletal Tissue Engineering

  • Gianluca Cidonio
  • Michael Glinka
  • Yang-Hee Kim
  • Jonathan I. Dawson
  • Richard O. C. OreffoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2147)


Biofabrication is revolutionizing substitute tissue manufacturing. Skeletal stem cells (SSCs) can be blended with hydrogel biomaterials and printed to form three-dimensional structures that can closely mimic tissues of interest. Our bioink formulation takes into account the potential for cell printing including a bioink nanocomposite that contains low fraction polymeric content to facilitate cell encapsulation and survival, while preserving hydrogel integrity and mechanical properties following extrusion. Clay inclusion to the nanocomposite strengthens the alginate-methylcellulose network providing a biopaste with unique shear-thinning properties that can be easily prepared under sterile conditions. SSCs can be mixed with the clay-based paste, and the resulting bioink can be printed in 3D structures ready for implantation. In this chapter, we provide the methodology for preparation, encapsulation, and printing of SSCs in a unique clay-based bioink.

Key words

Biofabrication Bioink Clay Laponite Scaffolds Bone repair Skeletal stem cell 



The authors would like to thank Prof. Michael Gelinsky (TU Dresden) for useful discussions and fruitful collaborations over the last 3 years, Prof. Shoufeng Yang (KU Leuven) for discussions and access to the extrusion bioprinter, and Dr. Stuart Lanham for useful discussions on methods. This work was supported by grants from the Biotechnology and Biological Sciences Research Council UK (BB/ L00609X and BB/LO21072/1) and University of Southampton IfLS, FortisNet and Postgraduate awards to ROCO.


  1. 1.
    Carretero MI, Pozo M (2009) Clay and non-clay minerals in the pharmaceutical industry. Part I. Excipients and medical applications. Appl Clay Sci 46:73–80CrossRefGoogle Scholar
  2. 2.
    Carretero MI, Pozo M (2010) Clay and non-clay minerals in the pharmaceutical and cosmetic industries Part II. Active ingredients. Appl Clay Sci 47:171–181CrossRefGoogle Scholar
  3. 3.
    Dawson JI, Oreffo ROC (2013) Clay: New opportunities for tissue regeneration and biomaterial design. Adv Mater 25:4069–4086CrossRefGoogle Scholar
  4. 4.
    Ruzicka B, Zaccarelli E (2011) A fresh look at the Laponite phase diagram. Soft Matter 7:1268–1286CrossRefGoogle Scholar
  5. 5.
    Kroon M, Vos WL, Wegdam GH (1998) Structure and formation of a gel of colloidal disks. Int J Thermophys 19:887–894CrossRefGoogle Scholar
  6. 6.
    Abou B, Bonn D, Meunier J (2001) Aging dynamics in a colloidal glass. Phys Rev E Stat Phys Plasmas Fluids Relat Interdiscip Topics 64:6Google Scholar
  7. 7.
    Pignon F, Magnin A, Piau JM (1998) Thixotropic behavior of clay dispersions: combinations of scattering and rheometric techniques. J Rheol 42:1349–1373CrossRefGoogle Scholar
  8. 8.
    Dawson JI, Kanczler JM, Yang XB et al (2011) Clay gels for the delivery of regenerative microenvironments. Adv Mater 23:3304–3308CrossRefGoogle Scholar
  9. 9.
    Carrow JK, Cross LM, Reese RW et al (2018) Widespread changes in transcriptome profile of human mesenchymal stem cells induced by two-dimensional nanosilicates. Proc Natl Acad Sci U S A 115(17):E3905–E3913CrossRefGoogle Scholar
  10. 10.
    Hölzl K, Lin S, Tytgat L et al (2016) Bioink properties before, during and after 3D bioprinting. Biofabrication 8:032002CrossRefGoogle Scholar
  11. 11.
    Gibbs DMR, Black CRM, Hulsart-Billstrom G et al (2016) Bone induction at physiological doses of BMP through localization by clay nanoparticle gels. Biomaterials 99:16–23CrossRefGoogle Scholar
  12. 12.
    Liu X, Bhatia SR (2015) Laponite® and Laponite®-PEO hydrogels with enhanced elasticity in phosphate-buffered saline. Polym Adv Technol 26:874–879CrossRefGoogle Scholar
  13. 13.
    Viseras C, Aguzzi C, Cerezo P et al (2008) Biopolymer–clay nanocomposites for controlled drug delivery. Mater Sci Technol 24:1020–1026CrossRefGoogle Scholar
  14. 14.
    Gaharwar AK, Schexnailder PJ, Kline BP et al (2011) Assessment of using Laponite cross-linked poly(ethylene oxide) for controlled cell adhesion and mineralization. Acta Biomater 7:568–577CrossRefGoogle Scholar
  15. 15.
    Haraguchi K, Takehisa T, Ebato M (2006) Control of cell cultivation and cell sheet detachment on the surface of polymer/clay nanocomposite hydrogels. Biomacromolecules 7:3267–3275CrossRefGoogle Scholar
  16. 16.
    Ahlfeld T, Cidonio G, Kilian D et al (2017) Development of a clay based bioink for 3D cell printing for skeletal application. Biofabrication 9:034103CrossRefGoogle Scholar
  17. 17.
    Billiet T, Gevaert E, De Schryver T et al (2014) The 3D printing of gelatin methacrylamide cell-laden tissue-engineered constructs with high cell viability. Biomaterials 35:49–62CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  • Gianluca Cidonio
    • 1
  • Michael Glinka
    • 1
  • Yang-Hee Kim
    • 1
  • Jonathan I. Dawson
    • 1
  • Richard O. C. Oreffo
    • 1
    Email author
  1. 1.Bone and Joint Research Group, Centre for Human Development, Stem Cells and Regeneration, Institute of Developmental SciencesUniversity of SouthamptonSouthamptonUK

Personalised recommendations