Polyelectrolyte gelatin-chitosan hydrogel optimized for 3D bioprinting in skin tissue engineering

VIEWS - 2067 (Abstract) 1469 (PDF)
Wei Long Ng, Wai Yee Yeong, May Win Naing


Bioprinting is a promising automated platform that enables the simultaneous deposition of multiple types of cells and biomaterials to fabricate complex three-dimensional (3D) tissue constructs. Most of the previous bioprinting works focused on collagen-based biomaterial, which has poor printability and long crosslinking time. This posed a immerse challenge to create a 3D construct with pre-determined shape and configuration. There is a need for a functional material with good printability in order to fabricate a 3D skin construct. Recently, the use of chitosan for wound healing applications has attracted huge attention due to its attractive traits such as its antimicrobial properties and ability to trigger hemostasis. In this paper, we report the modification of chitosan-based biomaterials for functional 3D bioprinting. Modification to the chitosan was carried out via the oppositely charged functional groups from chitosan and gelatin at a specific pH of ~pH 6.5 to form polyelectrolyte complexes. The polyelectrolyte hydrogels were evaluated in terms of chemical interactions within polymer blend, rheological properties (viscosities, storage and loss modulus), printing resolution at varying pressures and feed rates and biocompatibility. The chitosan-based hydrogels formulated in this work exhibited good printability at room temperature, high shape fidelity of the printed 3D constructs and good biocompatibility with fibroblast skin cells.


3D printing; bioprinting; rapid prototyping; additive manufacturing; skin tissue engineering

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Lanza R, Langer R and Vacanti J P, 2011, Principles of Tissue Engineering, 4th edn, Academic Press, San Diego.

Watt F M and Fujiwara H, 2011, Cell-extracellular matrix interactions in normal and diseased skin. Cold Spring Harbor Perspectives in Biology, vol.3(4): a005124. http://dx.doi.org/10.1101/cshperspect.a005124

Murphy S V and Atala A, 2014, 3D bioprinting of tissues and organs, Nature biotechnology, vol.32: 773–785. http://dx.doi.org/10.1038/nbt.2958

Ozbolat I T and Yu Y, 2013, Bioprinting towards organ fabrication: Challenges and future trends. IEEE Transactions on Bio-Medical Engineering, vol.60(3): 691–693. http://dx.doi.org/10.1109/TBME.2013.2243912

Lee V, Singh G, Trasatti C, et al., 2013, Design and fabrication of human skin by three-dimensional bioprinting. Tissue Engineering Part C: Methods, vol.20(6): 473–484. http://dx.doi.org/10.1089/ten.TEC.2013.0335

Koch L, Deiwick A, Schlie S, et al., 2012, Skin tissue generation by laser cell printing. Biotechnology and Bioengineering, vol.109(7): 1855–1863. http://dx.doi.org/10.1002/bit.24455

Pereira R F, Barrias C C, Granja P L, et al., 2013, Advanced biofabrication strategies for skin regeneration and repair. Nanomedicine, vol.8(4): 603–621.


Cui X, Breitenkamp K, Finn M, et al., 2012, Direct human cartilage repair using three-dimensional bioprinting technology. Tissue Engineering Part A, vol.18(11-12): 1304–1312. http://dx.doi.org/10.1089/ten.tea.2011.0543

Kundu J, Shim J H, Jang J, et al., 2013, An additive manufacturing-based PCL alginate-chondrocyte bioprinted scaffold for cartilage tissue engineering. Journal of Tissue Engineering and Regenerative Medicine, vol.9(11): 1286–1297. http://dx.doi.org/10.1002/term.1682

Chang R, Emami K, Wu H, et al., 2010, Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model. Biofabrication, vol.2(4): 045004. http://dx.doi.org/10.1088/1758-5082/2/4/045004

Binder K W, Zhao W X, Aboushwareb T, et al., 2010, In situ bioprinting of the skin for burns. Journal of the American College of Surgeons, vol.211(3): S76. http://dx.doi.org/10.1016/j.jamcollsurg.2010.06.198

Tobin D J, 2006, Biochemistry of human skin—our brain on the outside. Chemical Society Reviews, vol.35(1): 52–67. http://dx.doi.org/10.1039/B505793K

Ng W L, Yeong W Y and Naing M W, 2015, Cellular approaches to tissue-engineering of skin: A review. Journal of Tissue Science & Engineering, vol.6: 150. http://dx.doi.org/10.4172/2157-7552.1000150

Michael S, Sorg H, Peck C-T, et al., 2013, Tissue engineered skin substitutes created by laser-assisted bioprinting form skin-like structures in the dorsal skin fold chamber in mice. Plos One, vol.8: e57741. http://dx.doi.org/10.1371/journal.pone.0057741

Lee W, Debasitis J C, Lee V K, et al., 2009, Multi-layered culture of human skin fibroblasts and keratinocytes through three-dimensional freeform fabrication. Biomaterials, vol.30(8): 1587–1595. http://dx.doi.org/10.1016/j.biomaterials.2008.12.009

Lee W, Lee V, Polio S, et al., 2010, On-demand three-dimensional freeform fabrication of multi-layered hydrogel scaffold with fluidic channels. Biotechnology and Bioengineering, vol.105(6): 1178–1186. http://dx.doi.org/10.1002/bit.22613

Weber L, Kirsch E, Müller P, et al., 1984, Collagen type distribution and macromolecular organization of connective tissue in different layers of human skin. Journal of Investigative Dermatology, vol.82(2): 156–160. http://dx.doi.org/10.1111/1523-1747.ep12259720

Muzzarelli R A, 2009, Chitins and chitosans for the re-pair of wounded skin, nerve, cartilage and bone. Carbohydrate Polymers, vol.76(2): 167–182. http://dx.doi.org/10.1016/j.carbpol.2008.11.002

Dash M, Chiellini F, Ottenbrite R M, et al., 2011, Chitosan—A versatile semi-synthetic polymer in biomedical applications. Progress in Polymer Science, vol.36(8): 981–1014. http://dx.doi.org/10.1016/j.progpolymsci.2011.02.001

Jayakumar R, Prabaharan M, Kumar P S, et al. 2011, Biomaterials based on chitin and chitosan in wound dressing applications. Biotechnology Advances, vol.29(3): 322–337. http://dx.doi.org/10.1016/j.biotechadv.2011.01.005

Rinaudo M, 2006, Chitin and chitosan: Properties and applications. Progress in Polymer Science, vol.31(7): 603–632. http://dx.doi.org/10.1016/j.progpolymsci.2006.06.001

Kim I Y, Seo S J, Moon H S, et al., 2008, Chitosan and its derivatives for tissue engineering applications. Biotechnology Advances, vol.26(1): 1–21. http://dx.doi.org/10.1016/j.biotechadv.2007.07.009

Ueno H, Nakamura F, Murakami M, et al., 2001, Evaluation effects of chitosan for the extracellular matrix production by fibroblasts and the growth factors produc-tion by macrophages. Biomaterials, vol.22(15): 2125– 2130. http://dx.doi.org/10.1016/S0142-9612(00)00401-4

Kong M, Chen X G, Xing K, et al., 2010, Antimicrobial properties of chitosan and mode of action: A state of the art review. International Journal of Food Microbiology, vol.144(1): 51–63. http://dx.doi.org/10.1016/j.ijfoodmicro.2010.09.012

Cho Y-W, Jang J, Park C R, et al., 2000, Preparation and solubility in acid and water of partially deacetylated chitins. Biomacromolecules, vol.1(4): 609–614. http://dx.doi.org/10.1021/bm000036j

Geng L, Feng W, Hutmacher D W, et al., 2005, Direct writing of chitosan scaffolds using a robotic system. Rapid Prototyping Journal, vol.11(2): 90–97. http://dx.doi.org/10.1108/13552540510589458

Ng W L, Yeong W Y and Naing M W, 2014, Potential of bioprinted films for skin tissue engineering, in Proceedings of the 1st International Conference on Progress in Additive Manufacturing (eds C K Chua, W Y Yeong, M J Tan and E Liu).

Malafaya P B, Silva G A and Reis R L, 2007, Natural–origin polymers as carriers and scaffolds for biomolecules and cell delivery in tissue engineering applica-tions. Advanced Drug Delivery Reviews, vol.59(4–5): 207–233. http://dx.doi.org/10.1016/j.addr.2007.03.012

Huang Y, Onyeri S, Siewe M, et al., 2005, In vitro characterization of chitosan–gelatin scaffolds for tissue engineering. Biomaterials, vol.26(36): 7616–7627. http://dx.doi.org/10.1016/j.biomaterials.2005.05.036

Yin Y, Li Z, Sun Y, et al., 2005, A preliminary study on chitosan/gelatin polyelectrolyte complex formation. Journal of Materials Science, vol.40(17): 4649–4652. http://dx.doi.org/10.1007/s10853-005-3929-9

Mao J, Zhao L G, Yin Y J, et al., 2003, Structure and properties of bilayer chitosan–gelatin scaffolds. Biomaterials, vol.24(6): 1067–1074. http://dx.doi.org/10.1016/S0142-9612(02)00442-8

Mao J , Zhao L, de Yao K, et al., 2003, Study of novel chitosan-gelatin artificial skin in vitro. Journal of Biomedical Materials Research Part A, vol.64A(2): 301–308. http://dx.doi.org/10.1002/jbm.a.10223

Pereda M, Ponce A, Marcovich N, et al., 2011, Chitosan-gelatin composites and bi-layer films with potential antimicrobial activity. Food Hydrocolloids, vol.25(5): 1372–1381. http://dx.doi.org/10.1016/j.foodhyd.2011.01.001

Wang X, Yu X, Yan Y, et al., 2008, Liver tissue responses to gelatin and gelatin/chitosan gels. Journal of Biomedical Materials Research Part A, vol.87(1): 62–68. http://dx.doi.org/10.1002/jbm.a.31712

Nguyen K T and West J L, 2002, Photopolymerizable hydrogels for tissue engineering applications. Biomaterials, vol.23(22): 4307–4314. http://dx.doi.org/10.1016/S0142-9612(02)00175-8

Cheng M, Deng J, Yang F, et al., 2003, Study on physical properties and nerve cell affinity of composite films from

chitosan and gelatin solutions. Biomaterials, vol.24(17): 2871–2880. http://dx.doi.org/10.1016/S0142-9612(03)00117-0

Yin Y J, Yao K D, Cheng G X, et al., 1999, Properties of polyelectrolyte complex films of chitosan and gelatin. Polymer International, vol.48(6): 429–432. http://dx.doi.org/10.1002/(SICI)1097-0126(199906)48:6<429::AID-PI160>3.0.CO;2-1

Das S, Pati F, Choi Y J, et al., 2015, Bioprintable, cell-laden silk fibroin–gelatin hydrogel supporting multilineage differentiation of stem cells for fabrication of three-dimensional tissue constructs. Acta Biomaterialia, vol.11: 233–246. http://dx.doi.org/10.1016/j.actbio.2014.09.023

Swope V B, Supp A P and Boyce S T, 2002, Regulation of cutaneous pigmentation by titration of human melanocytes in cultured skin substitutes grafted to athymic mice. Wound Repair and Regeneration, vol.10(6): 378–386. http://dx.doi.org/10.1046/j.1524-475X.2002.10607.x

Mao J S, Cui Y L, Wang X H, et al. 2004, A preliminary study on chitosan and gelatin polyelectrolyte complex cytocompatibility by cell cycle and apoptosis analysis. Biomaterials, vol.25(18): 3973–3981. http://dx.doi.org/10.1016/j.biomaterials.2003.10.080

MacNeil S, 2007, Progress and opportunities for tissue-engineered skin. Nature, vol.445: 874–880. http://dx.doi.org/10.1038/nature05664

DOI: http://dx.doi.org/10.18063/IJB.2016.01.009


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