Interplay between crosslinking and ice nucleation controls the porous structure of freeze-dried hydrogel scaffolds
Open accessAbstract
Freeze-drying is a process of choice to texture hydrogel scaffolds with pores formed by an ice-templating mechanism. Using state-of-the-art microscopies (cryo-EBSD, μCT, CLSM), this work evidences and quantifies the effect of crosslinking and ice nucleation temperature on the porous structure of thin hydrogel scaffolds freeze-dried at a low cooling rate. We focused on a polysaccharide-based hydrogel and developed specific protocols to monitor or trigger ice nucleation for this study. At a fixed number of intermolecular crosslinks per primary molecule (p = 5), the mean pore size in the dry state decreases linearly from 240 to 170 μm, when ice nucleation temperature decreases from -6 °C to -18 °C. When ice nucleation temperature is fixed at -10 °C, the mean pore size decreases from 250 to 150 μm, as the crosslinking degree increases from p = 3 to p = 7. Scaffold infiltration ability was quantified with synthetic microspheres. The seeding efficiency was assessed with MC3T3-E1 individual cells and HepaRG™ spheroids. These data collapse into a single master curve that exhibits a sharp transition from 100 % to 0 %-efficiency as the entity diameter approaches the mean pore size in the dry state. Altogether, we can thus precisely tune the porosity of these 3D materials of interest for 3D cell culture and cGMP production for tissue engineering.
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During freeze-drying, as ice crystals grow, aNFC and AP molecules are expelled from the freezing front and concentrated in the non-frozen intergranular regions; polymer chains are forced into closer proximity until ice growth ceases.
“During freeze-drying, as ice crystals grow, aNFC and AP molecules are expelled from the freezing front and concentrated in the non-frozen intergranular regions. Through this cryo-concentration process, polymer chains are forced into closer proximity until ice growth ceases [40].”
From Amidated Pectin/Nanocellulose Hybrid Cryogel System with a pH-Responsive Release Profile for Small Intestinal Delivery (2025) · cites this paper as [40]
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- Authors
- Grenier, J., Duval, H., Lv, P., Barou, F., Le Guilcher, C., Aid, R., David, B., & Letourneur, D.
- Affiliations
- Centre National de la Recherche Scientifique (FR)
- Université Paris-Saclay (FR)
- Inserm (FR)
- Corresponding author
- Duval, Hervé
- Venue
- Biomaterials Advances
- Publisher
- Elsevier BV
- Published
- 2022-08
- Volume
- 139
- Pages
- 212973
- ISSN
- 2772-9508 (ISSN-L)
- OpenAlex
- W4282009019
- Cited by
- 72
- References
- 63
- Citations by year
- 2026: 30 · 2025: 25 · 2024: 9 · 2023: 5 · 2022: 3
- FWCI
- 3.44
- Type
- journal article
- Language
- en
- Open access
- green
- hal.science
- Licence
- Locations
- Biomaterials Advances (publishedVersion)
- PubMed (publishedVersion)
- HAL (submittedVersion, other-oa)
- HAL (submittedVersion)
- Funders
- Agence Nationale de la Recherche (ANR-11-IDEX-0003, ANR-10-LABX-0032, ANR-16-RHUS-0005)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Hydrogels: synthesis, properties, applications
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- freeze-drying, hydrogel scaffolds, ice nucleation, crosslink density, porous structure, pore size, polysaccharide hydrogels, freeze casting, seeding efficiency, 3D cell culture, tissue engineering, MC3T3-E1 cells, porosity control
- MeSH terms
- Freeze Drying, Ice, Porosity, Hydrogels, Tissue Engineering
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