AccScience Publishing / IJB / Online First / DOI: 10.36922/IJB025100082
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RESEARCH ARTICLE

High-precision depth-controlled laser bioprinting of cells in extracellular matrix for three-dimensional structures

Stavroula Elezoglou1 ,  Antonis Hatziapostolou1 ,  Kyriakos Giannakopoulos1 ,  Ioanna Zergioti1,2*
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1 Department of Physics, Institute of Communication and Computer Systems (ICCS), National Technical University of Athens, Athens, Greece
2 PhosPrint P.C., Lefkippos Technology Park, Athens, Greece
IJB, 025100082 https://doi.org/10.36922/IJB025100082
Received: 4 March 2025 | Accepted: 24 March 2025 | Published online: 24 March 2025
(This article belongs to the Special Issue Advances in 3D Bioprinting)
© 2025 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

Bioprinting is an emerging additive manufacturing process that offers great potential for fabricating living tissue by precisely printing cells and biomaterials onto various substrates. This technique can imitate native tissue functions, enabling clinical trials to explore new pathways for regenerative medicine. Among various bioprinting techniques, laser-induced forward transfer (LIFT) offers a high spatial resolution, accurate and controlled bio-ink deposition, and high post-printing cell viability. Effective bioprinting requires a deep understanding of material properties, especially the rheological behavior of bio-inks, which is critical for achieving the desired outcomes. Rheological characterization of these materials is essential for understanding their behavior under bioprinting conditions. The LIFT technique utilizes a wide range of soft biomaterials, generating printed structures containing cells, which proliferate for several days post-printing. These biomaterials can be controllably deposited in a variety of substrates. In this study, two cell-laden bio-inks with low and high number cell densities were printed at controlled depths within an extracellular matrix (ECM) by adjusting the laser energy. This process allows precise immobilization of cells at desired depths within the ECM using light and a proper optical setup. The rheological behavior of all bio-inks was analyzed using a microfabricated rheometer–viscometer on a chip. To investigate the transfer dynamics, a high-speed camera was integrated into the LIFT setup, monitoring the immobilization phenomenon within the ECM, and highlighting important characteristics of the jet propagations during printing. The morphological characteristics of the two sequential and distinct cell-laden jets were examined in detail during the printing process. This study showcases the ability to precisely deposit cells up to 2.5 mm deep within a soft matrix substrate, fabricating any desired cell-laden architecture for bio-engineering applications.  

Graphical abstract
Keywords
Bioprinting in-depth
Bio-fabrication
Laser bioprinting
Laser-induced forward transfer
Rheology
Three-dimensional structures
Funding
This work was funded by the EU Horizon 2020 FET program UroPrint under grant agreement No 964883.
Conflict of interest
Ioanna Zergioti serves as the Editorial Board Member and guest editor 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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