journal article·2025-07Open access

Utilising design of experiment to design an optimised bioink for 3D bioprinting

Alice Upton, Athina Mylona, Georgina Zimbitas

Journal of Materials Science· Volume 60· Issue 25

Abstract

Abstract The advancement of bioinks for 3D bioprinting is vital for tissue engineering, requiring precise tailoring of rheological and structural properties. This study employs integration of rheological analysis with a design of experiment (DoE) approach, with the aim being the optimisation of bioink formulations comprising of hyaluronic acid, sodium alginate, and dextran-40. A factorial DoE identified sodium alginate as the primary determinant of the bioinks' viscosity, while the mixture DoE established an optimal formulation with a viscosity of 3.275 Pa·s, matching the viscosity of the commercial benchmark. Rheological assessments confirmed the optimised bioink's shear-thinning properties and structural integrity, essential for printability and cellular support. Capability analysis of multiple batches demonstrated process reliability, whereby viscosities were consistently within defined boundaries, emphasising the robustness of the DoE-guided formulation process. This research highlights the potential of combining statistical and rheological methodologies to develop bioinks tailored for specific tissue applications, paving the way for improved 3D bioprinting outcomes.
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Research context

OpenAlex topics: 3D Printing in Biomedical Research · Additive Manufacturing and 3D Printing Technologies · Innovative Microfluidic and Catalytic Techniques Innovation

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

References (34)

Show source-provided references
1. Langer R, Vacanti JP (1993) Tissue engineering. Science. https://doi.org/10.1126/science.849352910.1126/science.8493529 ↗
2. Gao X, Ruzbarsky JJ, Layne JE, Xiao X, Huard J (2024) Stem cells and bone tissue engineering. Life (Basel) 14(3):287. https://doi.org/10.3390/life1403028710.3390/life14030287 ↗
3. Chung C, Burdick JA (2008) Engineering cartilage tissue. Adv Drug Deliv Rev 60(2):243–262. https://doi.org/10.1016/j.addr.2007.08.02710.1016/j.addr.2007.08.027 ↗
4. Kaul H, Ventikos Y (2015) On the genealogy of tissue engineering and regenerative medicine. Tissue Eng Part B Rev 21(2):203–217. https://doi.org/10.1089/ten.teb.2014.028510.1089/ten.teb.2014.0285 ↗
5. Gungor-Ozkerim PS, Inci I, Zhang YS, Khademhosseini A, Dokmeci MR (2018) Bioinks for 3D bioprinting: an overview. Biomater Sci 6(5):915–946. https://doi.org/10.1039/c7bm00765e10.1039/c7bm00765e ↗
6. Decante G, Costa JB, Silva-Correia J, Collins MN, Reis RL, Oliveira JM (2021) Engineering bioinks for 3D bioprinting. Biofabrication 13(3):032001. https://doi.org/10.1088/1758-5090/abec2c10.1088/1758-5090/abec2c ↗
7. Jawli A, Aldehani W, Nabi G, Huang Z (2024) Tissue-mimicking material fabrication and properties for multiparametric ultrasound phantoms: a systematic review. Bioengineering 11(6):620. https://doi.org/10.3390/bioengineering1106062010.3390/bioengineering11060620 ↗
8. Williams DF (2008) On the mechanisms of biocompatibility. Biomaterials 29(20):2941–2953. https://doi.org/10.1016/j.biomaterials.2008.04.02310.1016/j.biomaterials.2008.04.023 ↗
9. Schwab A, Levato R, D’Este M, Piluso S, Eglin D, Malda J (2020) Printability and shape fidelity of bioinks in 3D bioprinting. Chem Rev 120(19):11028–11055. https://doi.org/10.1021/acs.chemrev.0c0008410.1021/acs.chemrev.0c00084 ↗
10. Jo HJ, Kang MS, Kim JM et al (2024) Bioactive ions-loaded bioinks primed for 3dDprinting of artificial tissues. Biomed Mater Dev 2(2):811–833. https://doi.org/10.1007/s44174-023-00151-310.1007/s44174-023-00151-3 ↗
11. Chimene D, Kaunas R, Gaharwar AK (2020) Hydrogel bioink reinforcement for additive manufacturing: a focused review of emerging strategies. Adv Mater 32(1):1902026. https://doi.org/10.1002/adma.20190202610.1002/adma.201902026 ↗
12. Williams D, Thayer P, Martinez H, Gatenholm E, Khademhosseini A (2018) A perspective on the physical, mechanical and biological specifications of bioinks and the development of functional tissues in 3D bioprinting. Bioprinting 9:19–36. https://doi.org/10.1016/j.bprint.2018.02.00310.1016/j.bprint.2018.02.003 ↗
13. Pedroza-González SC, Rodriguez-Salvador M, Pérez-Benítez BE, Alvarez MM, de Santiago GT (2021) Bioinks for 3D bioprinting: a scientometric analysis of two decades of progress. Int J Bioprint. 7(2):333. https://doi.org/10.18063/ijb.v7i2.33710.18063/ijb.v7i2.337 ↗
14. Kaklamani G, Cheneler D, Grover LM, Adams MJ, Bowen J (2014) Mechanical properties of alginate hydrogels manufactured using external gelation. J Mech Behav Biomed Mater 36:135–142. https://doi.org/10.1016/j.jmbbm.2014.04.01310.1016/j.jmbbm.2014.04.013 ↗
15. Marques CF, Diogo GS, Pina S, Oliveira JM, Silva TH, Reis RL (2019) Collagen-based bioinks for hard tissue engineering applications: a comprehensive review. J Mater Sci: Mater Med 30(3):32. https://doi.org/10.1007/s10856-019-6234-x10.1007/s10856-019-6234-x ↗
16. Yang J, He H, Li D, Zhang Q, Xu L, Ruan C (2023) Advanced strategies in the application of gelatin-based bioink for extrusion bioprinting. Bio-des Manuf 6(5):586–608. https://doi.org/10.1007/s42242-023-00236-410.1007/s42242-023-00236-4 ↗
17. Burdick JA, Prestwich GD (2011) Hyaluronic acid hydrogels for biomedical applications. https://doi.org/10.1002/adma.201003963. Accessed 3 March 202510.1002/adma.201003963 ↗
18. Arif ZU, Khalid MY, Noroozi R, Sadeghianmaryan A, Jalalvand M, Hossain M (2022) Recent advances in 3D-printed polylactide and polycaprolactone-based biomaterials for tissue engineering applications. Int J Biol Macromol 218:930–968. https://doi.org/10.1016/j.ijbiomac.2022.07.14010.1016/j.ijbiomac.2022.07.140 ↗
19. Cui L, Tong W, Zhou H, Yan C, Chen J, Xiong D (2021) PVA-BA/PEG hydrogel with bilayer structure for biomimetic articular cartilage and investigation of its biotribological and mechanical properties. J Mater Sci 56(5):3935–3946. https://doi.org/10.1007/s10853-020-05467-910.1007/s10853-020-05467-9 ↗
20. Mendoza-Cerezo L, Jesús MRR, Macías-García A, Marcos-Romero AC, Díaz-Parralejo A (2023) Evolution of bioprinting and current applications. Int J Bioprint. 9(4):742. https://doi.org/10.18063/ijb.74210.18063/ijb.742 ↗
21. Paxton N, Smolan W, Böck T, Melchels F, Groll J, Jungst T (2017) Proposal to assess printability of bioinks for extrusion-based bioprinting and evaluation of rheological properties governing bioprintability. Biofabrication 9(4):044107. https://doi.org/10.1088/1758-5090/aa8dd810.1088/1758-5090/aa8dd8 ↗
22. Kiyotake EA, Douglas AW, Thomas EE, Nimmo SL, Detamore MS (2019) Development and quantitative characterization of the precursor rheology of hyaluronic acid hydrogels for bioprinting. Acta Biomater 95:176–187. https://doi.org/10.1016/j.actbio.2019.01.04110.1016/j.actbio.2019.01.041 ↗
23. Galocha-León C, Antich C, Voltes-Martínez A et al (2025) Human mesenchymal stromal cells-laden crosslinked hyaluronic acid-alginate bioink for 3D bioprinting applications in tissue engineering. Drug Deliv Transl Res 15(1):291–311. https://doi.org/10.1007/s13346-024-01596-910.1007/s13346-024-01596-9 ↗
24. Butler HM, Naseri E, MacDonald DS, Tasker RA, Ahmadi A (2021) Investigation of rheology, printability, and biocompatibility of N, O-carboxymethyl chitosan and agarose bioinks for 3D bioprinting of neuron cells. Materialia. https://doi.org/10.1016/j.mtla.2021.10116910.1016/j.mtla.2021.101169 ↗
25. Yang Y, Jia Y, Yang Q, Xu F (2022) Engineering bio-inks for 3D bioprinting cell mechanical microenvironment. Int J Bioprint. 9(1):632. https://doi.org/10.18063/ijb.v9i1.63210.18063/ijb.v9i1.632 ↗
26. Bercea M (2023) Rheology as a tool for fine-tuning the properties of printable bioinspired gels. Molecules 28(6):2766. https://doi.org/10.3390/molecules2806276610.3390/molecules28062766 ↗
27. Pössl A, Hartzke D, Schmidts TM, Runkel FE, Schlupp P (2021) A targeted rheological bioink development guideline and its systematic correlation with printing behavior. Biofabrication 13(3):035021. https://doi.org/10.1088/1758-5090/abde1e10.1088/1758-5090/abde1e ↗
28. Heathman TRJ, Webb WR, Han J et al (2014) Controlled production of poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) nanoparticles for targeted and sustained drug delivery. J Pharm Sci 103(8):2498–2508. https://doi.org/10.1002/jps.2403510.1002/jps.24035 ↗
29. Petta D, D’Amora U, Ambrosio L, Grijpma DW, Eglin D, D’Este M (2020) Hyaluronic acid as a bioink for extrusion-based 3D printing. Biofabrication 12(3):032001. https://doi.org/10.1088/1758-5090/ab875210.1088/1758-5090/ab8752 ↗
30. Schmid R, Schmidt SK, Detsch R et al (2022) A new printable alginate/hyaluronic acid/gelatin hydrogel suitable for biofabrication of in vitro and in vivo metastatic melanoma models. Adv Funct Mater 32(2):2107993. https://doi.org/10.1002/adfm.20210799310.1002/adfm.202107993 ↗
31. Zidarič T, Milojević M, Gradišnik L, StanaKleinschek K, Maver U, Maver T (2020) Polysaccharide-based bioink formulation for 3D bioprinting of an in vitro model of the human dermis. Nanomaterials (Basel) 10(4):733. https://doi.org/10.3390/nano1004073310.3390/nano10040733 ↗
32. Cavallo A, Al Kayal T, Mero A et al (2023) Marine collagen-based bioink for 3D bioprinting of a bilayered skin model. Pharmaceutics 15(5):1331. https://doi.org/10.3390/pharmaceutics1505133110.3390/pharmaceutics15051331 ↗
33. Gervasi V, Cullen S, McCoy T, Crean A, Vucen S (2019) Application of a mixture DOE for the prediction of formulation critical temperatures during lyophilisation process optimisation. Int J Pharm 572:118807. https://doi.org/10.1016/j.ijpharm.2019.11880710.1016/j.ijpharm.2019.118807 ↗
34. Avramova T, Vasileva D, Peneva T (2024) An overview of the basic concepts and terms related to manufacturing process capability evaluation. AIP Conf Proc 3104:020011. https://doi.org/10.1063/5.019883510.1063/5.0198835 ↗