Abstract
This research paper focuses on the use of the Freeform Reversible Embedding of Suspended Hydrogels (FRESH) printing method to fabricate complex structures with tunable elastic modulus and porosity. Currently, three-dimensional (3D) bioprinting has been widely used to construct hard tissues such as bone and cartilage. However, constructing soft tissues with complex structures remains a challenge. The FRESH printing method can overcome the effects of gravity on printed structures and is expected to become a key technique for preparing complex soft tissues. In this study, alginate and gelatin were used as ink substrates for cell-laden printing via the FRESH method. First, bioink with tunable porosity and mechanical properties was designed and prepared, and extrusion printing was performed in a gelatin particle support bath using the FRESH method. The rheological properties of the ink were measured using a rheometer to analyze printability, and the extrusion state of the ink was further analyzed through simulations. Moreover, mechanical test results and micro-morphology showed that the printed structures have tunable elastic modulus and porosity. By optimizing the FRESH process, a series of complex structures were printed. Finally, L929 fibroblasts were used for cell-laden printing, and the cells exhibited high viability and good proliferation characteristics during a 7-day culture. All results indicate that the designed bioink has good biological and mechanical properties, and further enhancement of cell viability can be achieved by adding cell-compatible materials, demonstrating great potential in soft tissue printing.
Cite this article (Click the link at the bottom to download the PDF)
Chen Z, Huang C, Liu H, et al., 2023. 3D bioprinting of complex biological structures with tunable elastic modulus and porosity using freeform reversible embedding of suspended hydrogels. Bio-des Manuf 6(5):550–562. https://doi.org/10.1007/s42242-023-00251-5
Article Guide
Figure 1 FRESH printing process for tissues with tunable elastic modulus and porosity. (a) Bioink monomer materials; (b) Preparation of bioink materials; (c) FRESH 3D bioprinting process; (d) Printed structure preserved in the support bath; (e) Scaffold released from the support bath at 37 °C; (f) Schematic of the microstructure of the hydrogel. FRESH: Freeform Reversible Embedding of Suspended Hydrogels
Figure 2 Rheological property testing of bioink. (a) Schematic of the rheological test; (b) Gelation dynamics curves of bioinks containing 2%, 4%, and 6% (0.02, 0.04, and 0.06 g/mL) gelatin during heating and cooling processes. Legend: Solid indicates G′, hollow indicates G′′, W indicates heating, C indicates cooling, and numbers indicate gelatin concentrations; (c) Three high (1000 s−1) and low (0.01 s−1) shear rate cycles at 37 °C; (d) Shear scanning experiments at 37 °C; (e) Velocity field of bioink flowing through the needle; (f) Shear stress at different radius positions of three ink concentrations in the needle
Figure 3 Micro-morphology of hydrogels at gelatin concentrations of (a) 2% (0.02 g/mL), (b) 4% (0.04 g/mL), (c) 6% (0.06 g/mL), and (d) 8% (0.08 g/mL)
Figure 4 FRESH printing of complex structures. (a) Repeated compression test on printed cylindrical tubes; (b) Infusion test on printed elongated tubes; (c) Design and print of solid “SDU” model, size 16.8 mm × 7.08 mm × 2.00 mm, scale: 1 mm
Figure 5 Cell immunofluorescence staining. On day 1 (a) and day 7 (b) of cell culture, cells were immunofluorescently stained using AM/PI. Scale bar: 200 μm. AM: Acetoxymethyl; PI: Propidium Iodide
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Bio-Design and Manufacturing (中文名《生物设计与制造》), abbreviated as BDM, is a professional English bimonthly journal sponsored by Zhejiang University, with editors-in-chief Academician Yang Huayong and Academician Cui Zhanfeng. Established in 2018, it has been indexed by SCI-E and other databases since 2019, and as of 2023, it has been changed to a bimonthly publication. The latest impact factor announced in 2023 is 7.9, ranking in the Q1 area of JCR, 14/96.
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