journal article·2025-11-28Open access

Monitoring Storage Stability of 3D Printed Hydrogels

Barbara Schmieg, Sarah Gretzinger, Gisela Guthausen, Jürgen Hubbuch

Applied Sciences· Volume 15· Issue 23

Abstract

Bioprinting of Advanced Therapy Medicinal Products offers promising new strategies for personalized medicine, but it requires comprehensive, non-destructive characterization and quality monitoring. To support patients with tailor-made constructs composed of hydrogels and cells derived from allogeneic donors or autologous samples, several challenges must be addressed—such as on-demand production, robust manufacturing, appropriate storage and logistics, and destruction-free quality control—before successful translation into clinical applications or pharmacy is possible. Although experience in cryo-preservation, blood banking, and organ donation helps to identify critical process parameters, detecting variations in manufacturing and ensuring product stability remain essential. Quality monitoring of 3D-printed objects before and after storage by magnetic resonance imaging (MRI) is complemented here by measurements of total mass and volume. These established methods provide rapid, non-destructive feedback and have well-characterized statistical limitations. Total mass can be assessed quickly; however, such integral measurements do not reveal information about internal structures. MRI, in contrast, offers detailed, spatially resolved insights. By combining these analytical modalities, we quantitatively analyzed the storage stability of 3D-printed hydrogels—without living cells in this study—in order to demonstrate and validate the analytical approach. We describe a workflow for measuring mass and geometry of 3D-printed hydrogel lattices before and after storage under varying process parameters. Critical quality attributes (cQAs), including overall and internal structural fidelity as well as mass conservation, were monitored. The presented workflow supports the development of cryopreservation protocols and has potential applications in biomaterial development for bioprinting and in quality assessment of tailor-made artificial tissues.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: 3D Printing in Biomedical Research · Microfluidic and Bio-sensing Technologies · Innovative Microfluidic and Catalytic Techniques Innovation

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Institute of Engineering in Life Sciences, Section IV: Molecular Separation Engineering, Karlsruhe Institute of Technology (KIT), D-76131 Karlsruhe, Germany

Institute of Functional Interfaces, Karlsruhe Institute of Technology (KIT), D-76131 Karlsruhe, Germany

Institute of Mechanical Process Engineering and Mechanics, Karlsruhe Institute of Technology (KIT), D-76131 Karlsruhe, Germany

Engler Bunte Institut Water Chemistry and Technology, Karlsruhe Institute of Technology (KIT), D-76131 Karlsruhe, Germany

References (26)

Show source-provided references
1. Manufacturing Road Map for Tissue Engineering and Regenerative Medicine Technologies10.5966/sctm.2014-0254 ↗
2. Decentralized manufacturing of cell and gene therapies: Overcoming challenges and identifying opportunities10.1016/j.jcyt.2017.07.005 ↗
3. Shipping and Logistics Considerations for Regenerative Medicine Therapies10.1093/stcltm/szab025 ↗
4. Challenges and advantages of cell therapy manufacturing under Good Manufacturing Practices within the hospital setting10.1016/j.copbio.2020.05.005 ↗
5. Disrupting vaccine logistics10.1093/inthealth/ihab010 ↗
6. Bioinks for Space Missions: The Influence of Long-Term Storage of Alginate-Methylcellulose-Based Bioinks on Printability as well as Cell Viability and Function10.1002/adhm.202300436 ↗
7. Time–Temperature Management Along the Food Cold Chain: A Review of Recent Developments10.1111/1541-4337.12269 ↗
8. Hydrogel-based encapsulation of biological, functional tissue: Fundamentals, technologies and applications10.1007/s00339-007-4270-8 ↗
9. Cryoreservation of alginate–fibrin beads involving bone marrow derived mesenchymal stromal cells by vitrification10.1016/j.biomaterials.2008.09.030 ↗
10. Impact of alginate concentration on the viability, cryostorage, and angiogenic activity of encapsulated fibroblasts10.1016/j.msec.2016.04.055 ↗
11. Hawkins, B., Loper, K., Abazari, A., Alder, B., Arcidiacono, J.A., Ballica, R., Brower, S., Elliott, J.T., Gayed, B., and Goh, C.W. (2021). Cryopreservation of Cells for Use in Cell Therapies, Gene Therapies, and Regenerative Medicine Manufacturing: An Introduction and Best Practices Approach on How to Prepare, Cryopreserve, and Recover Cells, Cell Lines, and Cell-Based Tissue Product (Standard No. ANSI/PDA Standard 02-2021).
12. On the study of the freeze–thaw thermal process of a biological system10.1016/j.applthermaleng.2009.07.001 ↗
13. Automated image processing as an analytical tool in cell cryopreservation for bioprocess development10.1007/s00449-019-02071-3 ↗
14. Spadiut, O., Gundinger, T., Pittermann, B., and Slouka, C. (2020). Spatially Resolved Effects of Protein Freeze-Thawing in a Small-Scale Model Using Monoclonal Antibodies. Pharmaceutics, 12.10.3390/pharmaceutics12040382 ↗
15. Weber, D., and Hubbuch, J. (2021). Temperature Based Process Characterization of Pharmaceutical Freeze-Thaw Operations. Front. Bioeng. Biotechnol., 9.10.3389/fbioe.2021.617770 ↗
16. Bioprinting Complex Cartilaginous Structures with Clinically Compliant Biomaterials10.1002/adfm.201503423 ↗
17. Schmieg, B., Gretzinger, S., Schuhmann, S., Guthausen, G., and Hubbuch, J. (2022). Magnetic resonance imaging as a tool for quality control in extrusion-based bioprinting. Biotechnol. J., 17.10.1002/biot.202100336 ↗
18. Gretzinger, S., Schmieg, B., Guthausen, G., and Hubbuch, J. (2022). Virtual Reality as Tool for Bioprinting Quality Inspection: A Proof of Principle. Front. Bioeng. Biotechnol., 10.10.3389/fbioe.2022.895842 ↗
19. Haas, S., Schmieg, B., Wendling, P., Guthausen, G., and Hubbuch, J. (2022). Magnetic Resonance Imaging: Time-Dependent Wetting and Swelling Behavior of an Auxetic Hydrogel Based on Natural Polymers. Polymers, 14.10.3390/polym14225023 ↗
20. Evaluation of 3D-printed silicone phantoms with controllable MRI signal properties
21. MR based magnetic susceptibility measurements of 3D printing materials at 3 Tesla10.1016/j.jmro.2023.100138 ↗
22. Callaghan, P.T. (1991). Principles of Nuclear Magnetic Resonance Microscopy, Oxford University Press.10.1093/oso/9780198539445.001.0001 ↗
23. Bernstein, M.A., King, K.F., and Zhou, X.J. (2004). Handbook of MRI Pulse Sequences, Academic Press, Elsevier Inc.10.1016/B978-012092861-3/50021-2 ↗
24. Blümich, B. (2000). NMR Imaging of Materials, Oxford University Press.
25. Modersitzki, J. (2003). Numerical Methods for Image Registration, Oxford University Press.10.1093/acprof:oso/9780198528418.001.0001 ↗
26. Špiclin, Ž., McClelland, J., Kybic, J., and Goksel, O. (2020). An Image Registration Framework for Discontinuous Mappings Along Cracks. Biomedical Image Registration, Springer. WBIR 2020; Lecture Notes in Computer Science.10.1007/978-3-030-50120-4 ↗