The development of cell-adhesive hydrogel for 3D printing

Kenichi Arai, Yoshinari Tsukamoto, Hirotoshi Yoshida, Hidetoshi Sanae, Tanveer Ahmad Mir, Shinji Sakai, Toshiko Yoshida, Motonori Okabe, Toshio Nikaido, Masahito Taya, Makoto Nakamura

Article ID: 76
Vol 2, Issue 2, 2016, Article identifier:153-162

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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.


biomaterials; 3D-bioprinter; biofabrication

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Hata K, 2007, Current issues regarding skin substitutes using living cells as industrial materials. Journal of Artificial Organs, vol.10(3): 129–132.

Langer R and Vacanti J P, 1993, Tissue engineering. Science, vol.260(5110): 920–926.

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.

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.

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.

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.

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.

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.

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.

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.

Malda J, Visser J, Melchels F P, et al., 2013, 25th anniversary article: engineering hydrogels for biofabrication. Advanced Materials, vol.25(36): 5011–5028.

Skardal A and Atala A, 2015, Biomaterials for integration with 3-D bioprinting. Annals of Biomedical Engineering, vol.43(3): 730–746.

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.

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.

Arai K, Iwanaga S, Toda H, et al., 2011, Three-dimensional inkjet biofabrication based on designed images. Biofabrication, vol.3(3): 034113.

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.

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.

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.

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.

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.

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.

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.

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.

Yamada K M and Olden K, 1978, Fibronectins — adhesive glycoproteins of cell surface and blood. Nature, vol.275: 179–184.

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.

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.

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.

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.

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.

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.



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