Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Suitability of Gelatin Methacrylate and Hydroxyapatite Hydrogels for 3D Bioprinted Bone Tissue

Version 1 : Received: 27 December 2023 / Approved: 27 December 2023 / Online: 27 December 2023 (11:01:23 CET)

A peer-reviewed article of this Preprint also exists.

Stolarov, P.; de Vries, J.; Stapleton, S.; Morris, L.; Martyniak, K.; Kean, T.J. Suitability of Gelatin Methacrylate and Hydroxyapatite Hydrogels for 3D-Bioprinted Bone Tissue. Materials 2024, 17, 1218. Stolarov, P.; de Vries, J.; Stapleton, S.; Morris, L.; Martyniak, K.; Kean, T.J. Suitability of Gelatin Methacrylate and Hydroxyapatite Hydrogels for 3D-Bioprinted Bone Tissue. Materials 2024, 17, 1218.

Abstract

Background: Complex bone defects are challenging to treat. Autografting is the gold standard for regenerating bone defects, however limitations include donor site morbidity and increased surgical complexity. Advancements in 3D bioprinting (3DBP) offer a promising alternative for viable bone grafts. Methods: Design of Experiments (DoE) was used to design 12 hydrogel bioinks of various Gelatin meth-acrylate/hydroxyapatite (GelMA/HA) concentrations. These bioinks were assessed for pipettability and equilibrium modulus. An optimal bioink was designed using the DoE data to produce the greatest stiffness while still being pipettable. Two bioinks, the DoE designed maximal stiff-ness and the experimentally defined maximal stiffness vs. a literature based control were then printed using a 3D bioprinter and assessed for print fidelity. The resulting hydrogels were combined with human bone-marrow-derived mesenchymal stromal cells (hMSCs) and evaluated for cell viability. Results: DoE ANOVA analysis indicated that the augmented 3 level factorial design model used was a good fit (p < 0.0001). Using the model, DoE correctly predicted that a composite hydrogel consisting of 12.3% GelMA, 15.7% HA, and 2% Gelatin would produce the maximum equilibrium modulus while still being pipettable. The hydrogel with the most optimal print fidelity was 10% GelMA, 2% HA, and 5% Gelatin. There were no significant differences in cell viability within hydrogels from day 2 to day 7 (p > 0.05). There was however a significantly lower cell viability in the gel composed of 12.3% GelMA, 15.7% HA, and 2% gelatin compared to the other gels with lower HA concentration (p < 0.05), showing that higher HA or print pressure may be cytotoxic within hydrogels. Conclusions: Extrusion-based 3DBP offers significant advantages for bone-tissue implants due to its high customizability. This study demonstrates it is possible to create printable bone-like grafts from GelMA and HA with increased HA levels, favorable mechanical properties (145kPa) and >80% cell viability.

Keywords

3D Bioprinting; Hydroxyapatite; GelMA; Bone; DoE; Design of Experiments; bone bioprinting

Subject

Biology and Life Sciences, Cell and Developmental Biology

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