Chapter 12
3D Bioprinting of Complex, Cell-laden Alginate Constructs
Atabak Ghanizadeh Tabriz, Dirk-Jan Cornelissen, and Wenmiao Shu
Abstract
Biofabrication has been receiving a great deal of attention in tissue engineering and regenerative medicine
either by manual or automated processes. Different automated biofabrication techniques have been used to
produce cell-laden alginate hydrogel structures, especially bioprinting approaches. These approaches have
been limited to 2D or simple 3D structures, however. In this chapter, a novel bioprinting technique is
disclosed for the production of more complex alginate hydrogel structures. This was achieved by dividing
the alginate hydrogel cross-linking process into three stages: primary calcium ion cross-linking for printability of the gel, secondary calcium ion cross-linking for rigidity of the alginate hydrogel immediately after
printing, and tertiary barium ion cross-linking for the long-term stability of the alginate hydrogel in the
culture medium.
Key words 3D bioprinting, Alginate hydrogel, Biofabrication, Bioextrusion
1 Introduction
Three-dimensional (3D) bioprinting has drawn great attention in
tissue engineering as an emerging technology during the past
decade for the regeneration and restoration of damaged or lost
tissues and organs [1–3] such as bones, skin, nose, and human ear
[4–7].
Several bioextrusion [8–10] printing methods have been developed to print living cells into a tissue or organ-like structure;
however, these structures are limited in complexity and scale.
The main challenge is balancing the printing conditions in such
a manner that bioscaffold material is viscous enough to generate a
rigid 3D structure while minimizing damage to the cells.
In this chapter, a new bioprinting technique using a modified
Fab@Home 3D printer for complex 3D alginate hydrogel structures is presented [11] as shown in Fig. 1. The technique uses a
three-step cross-linking process to ensure the printability, rigidity,
and stability of the gel that can be gentle to cells during the
bioprinting process. Alginate hydrogels were chosen because of
Alberto Rainer and Lorenzo Moroni (eds.), Computer-Aided Tissue Engineering: Methods and Protocols,
Methods in Molecular Biology, vol. 2147, https://doi.org/10.1007/978-1-0716-0611-7_12,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
143
3D Bioprinting of Complex, Cell-laden Alginate Constructs
Atabak Ghanizadeh Tabriz, Dirk-Jan Cornelissen, and Wenmiao Shu
Abstract
Biofabrication has been receiving a great deal of attention in tissue engineering and regenerative medicine
either by manual or automated processes. Different automated biofabrication techniques have been used to
produce cell-laden alginate hydrogel structures, especially bioprinting approaches. These approaches have
been limited to 2D or simple 3D structures, however. In this chapter, a novel bioprinting technique is
disclosed for the production of more complex alginate hydrogel structures. This was achieved by dividing
the alginate hydrogel cross-linking process into three stages: primary calcium ion cross-linking for printability of the gel, secondary calcium ion cross-linking for rigidity of the alginate hydrogel immediately after
printing, and tertiary barium ion cross-linking for the long-term stability of the alginate hydrogel in the
culture medium.
Key words 3D bioprinting, Alginate hydrogel, Biofabrication, Bioextrusion
1 Introduction
Three-dimensional (3D) bioprinting has drawn great attention in
tissue engineering as an emerging technology during the past
decade for the regeneration and restoration of damaged or lost
tissues and organs [1–3] such as bones, skin, nose, and human ear
[4–7].
Several bioextrusion [8–10] printing methods have been developed to print living cells into a tissue or organ-like structure;
however, these structures are limited in complexity and scale.
The main challenge is balancing the printing conditions in such
a manner that bioscaffold material is viscous enough to generate a
rigid 3D structure while minimizing damage to the cells.
In this chapter, a new bioprinting technique using a modified
Fab@Home 3D printer for complex 3D alginate hydrogel structures is presented [11] as shown in Fig. 1. The technique uses a
three-step cross-linking process to ensure the printability, rigidity,
and stability of the gel that can be gentle to cells during the
bioprinting process. Alginate hydrogels were chosen because of
Alberto Rainer and Lorenzo Moroni (eds.), Computer-Aided Tissue Engineering: Methods and Protocols,
Methods in Molecular Biology, vol. 2147, https://doi.org/10.1007/978-1-0716-0611-7_12,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
143