The development of cell-adhesive hydrogel for 3D printing

Kenichi Arai^1,2, Yoshinari Tsukamoto¹, Hirotoshi Yoshida¹, Hidetoshi Sanae¹, Tanveer Ahmad Mir¹, Shinji Sakai³, Toshiko Yoshida⁴, Motonori Okabe⁴, Toshio Nikaido⁴, Masahito Taya³, Makoto Nakamura^1*

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¹ Graduate School of Science and Engineering for Research, University of Toyama, Toyama, Japan

² Department of Regenerative Medicine and Biomedical Engineering, Saga University, Saga, Japan

³ Department of Materials Science and Engineering, Graduate School of Engineering Science, Osaka University, Osaka, Japan

⁴ Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan

IJB 2016, 2(2), 153–162; https://doi.org/10.18063/IJB.2016.02.002

Published: 22 June 2016

© 2016 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/ )

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Abstract

Biofabrication has gained tremendous attention for manufacturing functional organs or tissues. To fabricate functional organs or tissues, it is necessary to reproduce tissue-specific micro to macro structures. Previously, we developed a custom-made 3D-bioprinter with the capability to print and fabricate 3D complicated hydrogel structures composed of living cells. Through the gelation reaction, fine and complicated 3D gel structures can be fabricated via layer by layer printing. Alginate hydrogel has been used mainly due to its good fabricating properties. However, it is not a reliable platform for tissue regeneration because of its inadequate cell-adhesiveness. Therefore, our laboratory is interested to explore more suitable hydrogels for bioprinting and 3D tissue fabrication. In this study, we tried to fabricate 3D gel structures with enough cell-adhesive properties. We focused on hydrogel formation through enzymatic reaction by incorporating materials bearing phenolic hydroxyl moieties and horseradish peroxidase. We examined Alg-Ph and Alg-Ph/Gelatin-Ph gels. We used a mixed solution of applied materials as bioink and printed into H2O2 solution. We successfully fabricated the 3D gel sheet structures including fibroblasts cultures. Fibroblast proliferation and viability were also observed in the 3D gel sheet for more than one week. In conclusion, the hydrogel obtained through enzymatic reaction is a biocompatible bioink material which can be applied to fabricate 3D cell-adhesive gel structures using a 3D-bioprinter.

Keywords

biomaterials

3D-bioprinter

biofabrication

References

1. Hata K, 2007, Current issues regarding skin substitutes using living cells as industrial materials. Journal of Artificial Organs, vol.10(3): 129–132. http://dx.doi.org/10.1007/s10047-006-0371-y
2. Langer R and Vacanti J P, 1993, Tissue engineering. Science, vol.260(5110): 920–926. http://dx.doi.org/10.1126/science.8493529
3. Boyce S T, Kagan R J, Yakuboff K P, et al., 2002, Cultured skin substitutes reduce donor skin harvesting for closure of excised, full-thickness burns. Annals of Surgery, vol.235(2): 269–279. http://dx.doi.org/10.1097/00000658-200202000-00016
4. Du D, Furukawa K S and Ushida T, 2009, 3D culture of osteoblast-like cells by unidirectional or oscillatory flow for bone tissue engineering. Biotechnology and Bioen-gineering, vol.102(6): 1670–1678. http://dx.doi.org/10.1002/bit.22214
5. Banks-Schlegel S and Green H, 1981, Involucrin synthesis and tissue assembly by keratinocytes in natural and cultured human epithelia. The Journal of Cell Biol-ogy, vol.90(3): 732–737. http://dx.doi.org/10.1083/jcb.90.3.732
6. Nakamura S and Ijima H, 2013, Solubilized matrix derived from decellularized liver as a growth factor-immobilizable scaffold for hepatocyte culture. Journal of Bioscience and Bioengineering, vol.116(6): 746–753. http://dx.doi.org/10.1016/j.jbiosc.2013.05.031
7. Toyoda Y, Tamai M, Kashikura K, et al., 2012, Acetaminophen-induced hepatotoxicity in a liver tissue model consisting of primary hepatocytes assembling around an endothelial cell network. Drug Metabolism and Disposition, vol.40(1): 169–177.
http://dx.doi.org/10.1124/dmd.111.041137
8. Jiankang H, Dichen L, Yaxiong L, et al., 2009, Preparation of chitosan-gelatin hybrid scaffolds with well-organized microstructures for hepatic tissue engineering. Acta Biomaterialia, vol.5(1): 453–461. http://dx.doi.org/10.1016/j.actbio.2008.07.002
9. Tsuda Y, Kikuchi A, Yamato M, et al., 2006, Heterotypic cell interactions on a dually patterned surface. Biochemical and Biophysical Research Communications, vol.348(3): 937–944. http://dx.doi.org/10.1016/j.bbrc.2006.07.138
10. Yamada M, Utoh R, Ohashi K, et al., 2012, Controlled formation of heterotypic hepatic micro-organoids in anisotropic hydrogel microfibers for long-term preservation of liver-specific functions. Biomaterials, vol.33(33): 8304–8315. http://dx.doi.org/10.1016/j.biomaterials.2012.07.068
11. Malda J, Visser J, Melchels F P, et al., 2013, 25th anniversary article: engineering hydrogels for biofabrication. Advanced Materials, vol.25(36): 5011–5028. http://dx.doi.org/10.1002/adma.201302042
12. Skardal A and Atala A, 2015, Biomaterials for integration with 3-D bioprinting. Annals of Biomedical Engineering, vol.43(3): 730–746. http://dx.doi.org/10.1007/s10439-014-1207-1
13. Xu M, Wang X, Yan Y, et al., 2010, An cell-assembly derived physiological 3D model of the metabolic syndrome, based on adipose-derived stromal cells and a gela-tin/alginate/fibrinogen matrix. Biomaterials, vol.31(14): 3868–3877. http://dx.doi.org/10.1016/j.biomaterials.2010.01.111
14. Nishiyama Y, Nakamura M, Henmi C, et al., 2009, Development of a three-dimensional bioprinter: construction of cell supporting structures using hydrogel and state-of-the-art inkjet technology. Journal of Biomechanical Engineering, vol.131(3): 35001–35006. http://dx.doi.org/10.1115/1.3002759
15. Arai K, Iwanaga S, Toda H, et al., 2011, Three-dimensional inkjet biofabrication based on designed images. Biofabrication, vol.3(3): 034113. http://dx.doi.org/10.1088/1758-5082/3/3/034113
16. Onoe H, Okitsu T, Itou A, et al., 2013, Metre-long cell-laden microfibres exhibit tissue morphologies and functions. Nature Materials, vol.12(6): 584–590. http://dx.doi.org/10.1038/nmat3606
17. Li K, Qu X, Wang Y, et al., 2005, Improved performance of primary rat hepatocytes on blended natural polymers. Journal of Biomedical Materials Research Part A, vol.75(2): 268–274. http://dx.doi.org/10.1002/jbm.a.30415
18. Liu Y, Sakai S and Taya M, 2012, Production of endothelial cell-enclosing alginate-based hydrogel fibers with a cell adhesive surface through simultaneous cross-linking by horseradish peroxidase-catalyzed reaction in a hydrodynamic spinning process. Journal of Bioscience and Bioengineering, vol.114(3): 353–359. http://dx.doi.org/10.1016/j.jbiosc.2012.04.018
19. Liu Y, Sakai S, and Taya M, 2013, Impact of the composition of alginate and gelatin derivatives in bioconjugated hydrogels on the fabrication of cell sheets and spherical tissues with living cell sheaths. Acta Biomaterialia, vol.9(5): 6616–6623. http://dx.doi.org/10.1016/j.actbio.2013.01.037
20. Ashida T, Sakai S and Taya M, 2013, Competing two enzymatic reactions realizing one-step preparation of cell-enclosing duplex microcapsules. Biotechnology Progress, vol.29(6): 1528–1534. http://dx.doi.org/10.1002/btpr.1800
21. Sakai S, Ashida T and Ogino S, 2014, Horseradish peroxidase-mediated encapsulation of mammalian cells in hydrogel particles by dropping. Journal of Microencapsulation: Micro and Nano Carriers, vol.31(1): 100–104. http://dx.doi.org/10.3109/02652048.2013.808281
22. Sakai S and Kawakami K, 2007, Synthesis and characterization of both ionically and enzymatically cross-link-able alginate. Acta Biomaterialia, vol.3(4): 495–501. http://dx.doi.org/10.1016/j.actbio.2006.12.002
23. Sakai S, Hirose K, Taguchi K, et al. 2009, An injectable, in situ enzymatically gellable, gelatin derivative for drug delivery and tissue engineering. Biomaterials, vol.30(20): 3371–3377. http://dx.doi.org/10.1016/j.biomaterials.2009.03.030
24. Yamada K M and Olden K, 1978, Fibronectins — adhesive glycoproteins of cell surface and blood. Nature, vol.275: 179–184. http://dx.doi.org/10.1038/275179a0
25. Terranova V P, Rohrbach D H and Martin G R, 1980, Role of laminin in the attachment of PAM 212 (epithelial) cells to basement membrane collagen. Cell, vol.22(3): 719–726. http://dx.doi.org/10.1016/0092-8674(80)90548-6
26. Oda H, Yoshida Y, Kawamura A, et al. 2008, Cell shape, cell-cell contact, cell-extracellular matrix contact and cell polarity are all required for the maximum induction of CYP2B1 and CYP2B2 gene expression by phenobarbital in adult rat cultured hepatocytes. Biochemical Pharmacology, vol.75(5): 1209–1217. http://dx.doi.org/10.1016/j.bcp.2007.11.003
27. Sakai S, Liu Y, Mah E J, et al. 2013, Horseradish peroxidase/catalase-mediated cell-laden alginate-based hydrogel tube production in two-phase coaxial flow of aqueous solutions for filament-like tissues fabrication. Biofabrication, vol.5(1): 015012. http://dx.doi.org/10.1088/1758-5082/5/1/015012
28. Ogushi Y, Sakai S and Kawakami K, 2009, Phenolic hydroxy groups incorporated for the peroxidase-catalyzed gelation of a carboxymethylcellulose support: cellular adhesion and proliferation. Macromolecular Bioscience, vol.9(3): 262–267. http://dx.doi.org/10.1002/mabi.200800263
29. Kim J, Lee K W, Hefferan T E, et al., 2008, Synthesis and evaluation of novel biodegradable hydrogels based on poly(ethylene glycol) and sebacic acid as tissue engineering scaffolds. Biomacromolecules, vol.9(1): 149–157. http://dx.doi.org/10.1021/bm700924n
30. Frye C A, Wu X and Patrick Jr C W, 2005, Microvascular endothelial cells sustain preadipocyte viability under hypoxic conditions. In Vitro Cellular and Developmental Biology – Animal, vol.41(5): 160–164. http://dx.doi.org/10.1290/0502015.1

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