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RESEARCH ARTICLE

Internally-crosslinked alginate dialdehyde/ alginate/gelatin-based hydrogels as bioinks for prospective cardiac tissue engineering applications

Giovanni Paolo Stola1,2 ,  Camilla Paoletti1,2 ,  Letizia Nicoletti1,2 ,  Geo Paul3 ,  Claudio Cassino3 ,  Leonardo Marchese3 ,  Valeria Chiono1,2* ,  Elena Marcello1,2*
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1 Department of Mechanical and Aerospace Engineering, POLITO BioMedLab, Politecnico di Torino, Turin, Italy
2 Interuniversity Center for the Promotion of the 3Rs Principles in Teaching and Research (Centro 3R), Pisa, Italy
3 Department of Science and Technological Innovation and “Centro Interdisciplinare Nano-SiSTeMI”, Università del Piemonte Orientale, Alessandria, Italy
IJB, 4014 https://doi.org/10.36922/ijb.4014
Submitted: 21 June 2024 | Accepted: 18 September 2024 | Published: 18 September 2024
© 2024 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Cardiovascular diseases represent a global challenge due to heart-limited regenerative capabilities. 3D-bioprinted cell-laden constructs are a promising approach as cardiac patches or in vitro models. However, developing bioinks with optimal mechanical, rheological, and biological properties remains challenging. Although alginate (Alg)- based bioinks have been extensively explored, such hydrogels lack cell adhesion properties and degradability. Additionally, 3D Alg structures are usually obtained by microextrusion bioprinting, exploiting conventional external crosslinking methods, which introduce inhomogeneities and unpredictability in construct formation. This work exploits Alg internal ionic gelation mechanism to obtain homogeneous self-standing multilayered 3D-printed constructs without employing support baths or post-printing crosslinking treatments. Alg was blended with oxidized alginate (ADA) and gelatin (Gel) to achieve degradable and cell-adhesive hydrogels for cardiac tissue engineering. Firstly, ADA/Alg bioink composition was tailored to achieve cardiac tissue-like viscoelastic properties. Then, the amount of Gel in ADA/Alg hydrogels was optimized to support cell adhesion, producing shear thinning inks with tunable viscoelastic properties (storage modulus [G’]: 650–1300 Pa) and degradation profile (40–80% weight loss after 21 days in phosphate-buffered saline [PBS]) by varying Gel concentration. ADA/Alg/Gel hydrogels displayed shear thinning behavior, suitable for 3D bioprinting depending on the ink stabilization time, due to the gradual pH-triggered release of calcium ions over time. Adult human cardiac fibroblast (AHCF) and H9C2-laden ADA/Alg/Gel bioinks were successfully printed, producing scaffolds with high shape fidelity and good cell viability post-printing. Finally, the highest Gel content (25% [w/w]) supported cell adhesion after 24 h of incubation, displaying potential for cardiac tissue modeling. This research presents a comprehensive framework for advancing the design of bioink.  

 

Keywords
Alginate dialdehyde
Gelatin
Bioink
Internal gelation
In vitro cardiac models
Funding
This work was supported by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (BIORECAR, grant agreement number 772168).
Conflict of interest
Elena Marcello serves as the Editorial Board Member of the journal, but was not in any way involved in the editorial and peer-review process conducted for this paper, directly or indirectly. Other authors declare they have no competing interests.
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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing
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