AccScience Publishing / IJB / Volume 9 / Issue 4 / DOI: 10.18063/ijb.705
SHORT COMMUNICATION

Development and characterization of an automated active mixing platform for hydrogel bioink preparation

Jiannan Li1† Tara Shelby1† Hossein Vahid Alizadeh1 Hannah Shelby1 Yunzhi Peter Yang1,2,3*
Show Less
1 Department of Orthopedic Surgery, Stanford University, 240 Pasteur Drive, Stanford, CA94304, USA
2 Department of Materials Science and Engineering, Stanford University, 496 Lomita Mall, Stanford, CA94305, USA
3 Department of Bioengineering, Stanford University, 443 Via Ortega, Stanford, CA94305, USA
© Invalid date by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Abstract

Bioink preparation is an important yet challenging step for bioprinting with hydrogels, as it involves fast and homogeneous mixing of various viscous components. In this study, we have developed an automated active mixing platform (AAMP), which allows for high-quality preparation of hydrogel bioinks. The design of AAMP, adapted from syringe pumps, provides many advantages, including low cost, automated control, high precision, customizability, and great cytocompatibility, as well as the potential to intelligently detect the homogeneity. To demonstrate the capability of AAMP, mixing of different hydrogel components, including alginate and xanthan gum with and without Ca2+, alginate and Laponite, PEGDMA and xanthan gum, was performed to investigate an alginate hydrogel preparation process. Colorimetric analyses were carried out to evaluate the mixing outcome with AAMP. Result showed that AAMP can prepare homogeneous hydrogel mixing in a fast and automated fashion. A  multiphysics COMSOL simulation is carried out to further validate the results. Moreover, cell viability and proliferation study were performed in a cell encapsulation mixing experiment to validate the cytocompatibility of the AAMP. The AAMP has demonstrated great capability in hydrogel bioink preparation and could therefore holds great promise and wide applications in bioprinting and tissue engineering.

Keywords
Bioprinting
Hydrogel
Tissue engineering
Bioink
Biomaterials
References

1. Aswathy SH, Narendrakumar U, Manjubala I, 2020, Commercial hydrogels for biomedical applications. Heliyon, 6: e03719. https://doi.org/10.1016/j.heliyon.2020.e03719 

2. Billiet T, Vandenhaute M, Schelfhout J, et al., 2012, A review of trends and limitations in hydrogel-rapid prototyping for tissue engineering. Biomaterials, 33: 6020–6041. https://doi.org/10.1016/j.biomaterials.2012.04.050 

3. Khademhosseini A, Langer R, 2007, Microengineered hydrogels for tissue engineering. Biomaterials, 28: 5087–5092. https://doi.org/10.1016/j.biomaterials.2007.07.021 

4. Zhu J, Marchant RE, 2011, Design properties of hydrogel tissue-engineering scaffolds. Expert Rev Med Devices, 8: 607–626. https://doi.org/10.1586/erd.11.27 

5. Ji S, Guvendiren M, 2017, Recent advances in bioink design for 3D bioprinting of tissues and organs. Front Bioeng Biotechnol, 5: 23. https://doi.org/10.3389/fbioe.2017.00023 

6. Jia J, Richards DJ, Pollard S, et al., 2014, Engineering alginate as bioink for bioprinting. Acta Biomater, 10: 4323–4331. https://doi.org/10.1016/j.actbio.2014.06.034 

7. Kesti M, Muller M, Becher J, et al., 2015, A versatile bioink for three-dimensional printing of cellular scaffolds based on thermally and photo-triggered tandem gelation. Acta Biomater, 11: 162–172. https://doi.org/10.1016/j.actbio.2014.09.033
 
8. Kumar H, Sakthivel K, Mohamed MG, et al., 2021, Designing gelatin methacryloyl (GelMA)-based bioinks for visible light stereolithographic 3D biofabrication. Macromol Biosci, 21: e2000317. https://doi.org/10.1002/mabi.202000317 

9. Li N, Guo R, Zhang ZJ, 2021, Bioink formulations for bone tissue regeneration. Front Bioeng Biotechnol, 9: 630488. https://doi.org/10.3389/fbioe.2021.630488 

10. Naranda J, Bracic M, Vogrin M, et al., 2021, Recent advancements in 3D printing of polysaccharide hydrogels in cartilage tissue engineering. Materials (Basel), 14: 3977. https://doi.org/10.3390/ma14143977
 
11. Possl A, Hartzke D, Schmidts TM, et al., 2021, A targeted rheological bioink development guideline and its systematic correlation with printing behavior. Biofabrication, 13: 035021. https://doi.org/10.1088/1758-5090/abde1e 

12. Unagolla JM, Jayasuriya AC, 2020, Hydrogel-based 3D bioprinting: A comprehensive review on cell-laden hydrogels, bioink formulations, and future perspectives. Appl Mater Today, 18: 100479. https://doi.org/10.1016/j.apmt.2019.100479 

13. Yin J, Yan M, Wang Y, et al., 2018, 3D bioprinting of low-concentration cell-laden gelatin methacrylate (GelMA) bioinks with a two-step cross-linking strategy. ACS Appl Mater Interfaces, 10: 6849–6857. https://doi.org/10.1021/acsami.7b16059 

14. Zheng Z, Wu J, Liu M, et al., 2018, 3D bioprinting of self-standing silk-based bioink. Adv Healthc Mater, 7: e1701026. https://doi.org/10.1002/adhm.201701026 

15. Puertas-Bartolome M, Wlodarczyk-Biegun MK, Del Campo A, et al., 2020, 3D printing of a reactive hydrogel bio-ink using a static mixing tool. Polymers (Basel), 12: 1986. https://doi.org/10.3390/polym12091986
 
16. Tamayol A, Najafabadi AH, Aliakbarian B, et al., 2015, Hydrogel templates for rapid manufacturing of bioactive fibers and 3D constructs. Adv Healthc Mater, 4: 2146–2153. https://doi.org/10.1002/adhm.201500492 

17. Li Y, Yang HY, Lee DS, 2021, Advances in biodegradable and injectable hydrogels for biomedical applications. J Control Release, 330: 151–160. https://doi.org/10.1016/j.jconrel.2020.12.008 

18. Ma T, Lv L, Ouyang C, et al., 2021, Rheological behavior and particle alignment of cellulose nanocrystal and its composite hydrogels during 3D printing. Carbohydr Polyme, 253: 117217. https://doi.org/10.1016/j.carbpol.2020.117217 

19. Zhang W, Ma X, Li Y, et al., 2020, Preparation of smooth and macroporous hydrogel via hand-held blender for wound healing applications: In-vitro and in-vivo evaluations. Biomed Mater, 15: 055032. https://doi.org/10.1088/1748-605X/ab9d6f 

20. Mulakkal MC, Trask RS, Ting VP, et al., 2018, Responsive cellulose-hydrogel composite ink for 4D printing. Mater Des, 160: 108–118. https://doi.org/10.1016/j.matdes.2018.09.009 

21. Fu YC, Chen CH, Wang CZ, et al., 2013, Preparation of porous bioceramics using reverse thermo-responsive hydrogels in combination with rhBMP-2 carriers: In vitro and in vivo evaluation. J Mech Behav Biomed Mater, 27: 64–76. https://doi.org/10.1016/j.jmbbm.2013.06.009 

22. Müller M, Öztürk E, Arlov Ø, et al., 2017, Alginate sulfate-nanocellulose bioinks for cartilage bioprinting applications. Ann Biomed Eng, 45: 210–223. https://doi.org/10.1007/s10439-016-1704-5 

23. Apelgren P, Amoroso M, Säljö K, et al., 2018, Skin grafting on 3D bioprinted cartilage constructs in vivo. Plast Reconstr Surg Glob Open, 6: e1930. https://doi.org/10.1097/GOX.0000000000001930 

24. Apelgren P, Amoroso M, Lindahl A, et al., 2017, Chondrocytes and stem cells in 3D-bioprinted structures create human cartilage in vivo. PLoS One, 12: e0189428. https://doi.org/10.1371/journal.pone.0189428 

25. Cohen DL, Lo W, Tsavaris A, et al., 2011, Increased mixing improves hydrogel homogeneity and quality of three-dimensional printed constructs. Tissue Eng Part C Methods, 17: 239–248. https://doi.org/10.1089/ten.TEC.2010.0093 

26. Lowe SB, Tan VT, Soeriyadi AH, et al., 2014, Synthesis and high-throughput processing of polymeric hydrogels for 3D cell culture. Bioconjug Chem, 25: 1581–1601. https://doi.org/10.1021/bc500310v 

27. Rabanel JM, Hildgen P, 2004, Preparation of hydrogel hollow particles for cell encapsulation by a method of polyester core degradation. J Microencapsul, 21: 413–431. https://doi.org/10.1080/02652040410001729223 

28. Billiet T, Gevaert E, De Schryver T, et al., 2014, The 3D printing of gelatin methacrylamide cell-laden tissue-engineered constructs with high cell viability. Biomaterials, 35: 49–62. https://doi.org/10.1016/j.biomaterials.2013.09.078 

29. Wang LH, Ernst AU, An D, et al., 2021, A bioinspired scaffold for rapid oxygenation of cell encapsulation systems. Nat Commun, 12: 5846. https://doi.org/10.1038/s41467-021-26126-w 

30. Touani FK, Borie M, Azzi F, et al., 2021, Pharmacological preconditioning improves the viability and proangiogenic paracrine function of hydrogel-encapsulated mesenchymal stromal cells. Stem Cells Int, 2021: 6663467. https://doi.org/10.1155/2021/6663467 

31. Pan Z, Bui L, Yadav V, et al., 2021, Conformal single cell hydrogel coating with electrically induced tip streaming of an AC cone. Biomater Sci, 9: 3284–3292. https://doi.org/10.1039/d0bm02100h 

32. Karimi S, Bagher Z, Najmoddin N, et al., 2021, Alginate-magnetic short nanofibers 3D composite hydrogel enhances the encapsulated human olfactory mucosa stem cells bioactivity for potential nerve regeneration application. Int J Biol Macromol, 167: 796–806. https://doi.org/10.1016/j.ijbiomac.2020.11.199

33. Hwang DG, Jo Y, Kim M, et al., 2021, A 3D bioprinted hybrid encapsulation system for delivery of human pluripotent stem cell-derived pancreatic islet-like aggregates. Biofabrication, 14: 014101. https://doi.org/10.1088/1758-5090/ac23ac 

34. Ghasemi A, Akbari E, Imani R, 2021, An overview of engineered hydrogel-based biomaterials for improved β-cell survival and insulin secretion. Front Bioeng Biotechnol, 9: 662084. https://doi.org/10.3389/fbioe.2021.662084
 
35. Bonani W, Cagol N, Maniglio D, 2020, Alginate hydrogels: A tool for 3D cell encapsulation, tissue engineering, and biofabrication. Adv Exp Med Biol, 1250: 49–61. https://doi.org/10.1007/978-981-15-3262-7_4 

36. Garcia-Ochoa F, Santos VE, Casas JA, et al., 2000, Xanthan gum: Production, recovery, and properties. Biotechnol Adv, 18: 549–579. https://doi.org/10.1016/s0734-9750(00)00050-1 

37. Riaz T, Iqbal MW, Jiang B, et al., 2021, A review of the enzymatic, physical, and chemical modification techniques of xanthan gum. Int J Biol Macromol, 186: 472–489. https://doi.org/10.1016/j.ijbiomac.2021.06.196 

38. Wang L, Li W, Qin S, 2021, Three polymers from the sea: unique structures, directional modifications, and medical applications. Polymers (Basel), 13: 2482. https://doi.org/10.3390/polym13152482 

39. Van Hoorick J, Tytgat L, Dobos A, et al., 2019, (Photo-) crosslinkable gelatin derivatives for biofabrication applications. Acta Biomater, 97: 46–73. https://doi.org/10.1016/j.actbio.2019.07.035 

40. Chen H, Fei F, Li X, et al., 2021, A structure-supporting, self-healing, and high permeating hydrogel bioink for establishment of diverse homogeneous tissue-like constructs. Bioact Mater, 6: 3580–3595. https://doi.org/10.1016/j.bioactmat.2021.03.019 

41. Marques CF, Diogo GS, Pina S, et al., 2019, Collagen-based bioinks for hard tissue engineering applications: A comprehensive review. J Mater Sci Mater Med, 30: 32. https://doi.org/10.1007/s10856-019-6234-x 

42. Rajabi N, Rezaei A, Kharaziha M, et al., 2021, Recent advances on bioprinted gelatin methacrylate-based hydrogels for tissue repair. Tissue Eng Part A, 27: 679–702. https://doi.org/10.1089/ten.TEA.2020.0350 

43. Magalhaes LS, Santos FE, de Maria Vaz Elias C, et al., 2020, Printing 3D hydrogel structures employing low-cost stereolithography technology. J Funct Biomater, 11: 12. https://doi.org/10.3390/jfb11010012 

44. Acosta-Velez GF, Zhu TZ, Linsley CS, et al., 2018, Photocurable poly(ethylene glycol) as a bioink for the inkjet 3D pharming of hydrophobic drugs. Int J Pharm, 546: 145–153. https://doi.org/10.1016/j.ijpharm.2018.04.056

Share
Back to top
International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing