Biofabrication of vessel-like structures with alginate di-aldehyde—gelatin (ADA-GEL) bioink
One of the key challenges in the field of blood vessel engineering is the in vitro production of small and large diameter vessels. Considering that a combination of alginate di-aldehyde and gelatin (ADA-GEL) has been successfully applied for different biofabrication approaches, the aim of this study was to exploit ADA-GEL for the fabrication of vessel structures with diameters up to 4 mm. To explore plotting possibilities and to study the swelling behaviour, a library of vessel-like constructs with different diameters made from 2, 3 and 4% (w/v) alginate was created by using various hand-crafted double-needle extrusion systems. Vessel diameters were varied through changes of the double-needle core and outer diameters. A straightforward model for the production of vessel of different diameters from a variety of double-needle systems was established and vessel-constructs with diameters of up to 3.7 mm could be created. It was successfully demonstrated that an artificial vessel, consisting of an outer layer of 7.5% ADA50-GEL50 and an inner core of 3% gelatin, can support the proliferation and migration of an immobilized co-culture containing fibroblast (NHDF) and endothelial (HUVEC) cells. The openness and tightness of the hollow ADA-GEL structures were further confirmed by a dye injection test. Nanoindentation was performed to determine the Young’s modulus of the used materials. Cell vitality was proved after 1, 2 and 3 weeks of incubation. The results showed a nearly twofold increase of viable cells per week. Fluorescent images confirmed cell migration during the whole incubation time.
This work was supported by the German Research Foundation (DFG) within the collaborative research center TRR225 (project Nr. 326998133) (subprojects A01 and B06). Furthermore, this work was technically supported by Dr. Raminder Singh and PD Dr. Iwona Cicha from the Translational Research Center (TRC) of the University Medical Centre Erlangen. The authors would also like to thank Dr. T. Zehnder for his help with the used hydrogels and Ms. A. Grünewald for cell culturing.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 3.WHO (2018). The top 10 causes of death: World Heal Organ. http://www.who.int/mediacentre/factsheets/fs310/en/. Accessed on 17 Jan 2018.
- 10.Anatomie Blutgefäße (2017). http://www.physiologie-online.com/ana_site/organe-blutgefaesse.html. Accessed on 24 Oct 2017.
- 11.Aird WC. Phenotypic heterogeneity of the endothelium: I. structure, function, and mechanisms. Circ Res. 2007;100:158–73. https://doi.org/10.1161/01.RES.0000255691.76142.4a.CrossRefGoogle Scholar
- 12.Aird WC. Phenotypic heterogeneity of the endothelium: II. representative vascular beds. Circ Res. 2007;100:174–90. https://doi.org/10.1161/01.RES.0000255690.03436.ae.CrossRefGoogle Scholar
- 13.Nicolay N, Antwerpes F, Steudler U (2017). Tunica Adventitia. DocCheck. https://flexikon.doccheck.com/de/Tunica_adventitia.Accessed on 5 Sept 2017.
- 14.van den Berg F. Angewandte Physiologie: Band 1. Georg Thieme Verla; Stuttgart, Germany, 2003.Google Scholar
- 18.Lee KY, Mooney DJ. Alginate: properties and biomedical applications. Prog Polym Sci. 2012;37:106–26. https://doi.org/10.1016/j.progpolymsci.2011.06.003.CrossRefGoogle Scholar
- 19.Pawar SN, Edgar KJ. Alginate derivatization: a review of chemistry, properties and applications. Biomaterials. 2012;33:3279–305. https://doi.org/10.1016/j.biomaterials.2012.01.007.CrossRefGoogle Scholar
- 23.Sarker B, Papageorgiou DG, Silva R, Zehnder T, Gul-E-Noor F, Bertmer M. et al. Fabrication of alginate–gelatin crosslinked hydrogel microcapsules and evaluation of the microstructure and physico-chemical properties. J Mater Chem B. 2014;2:1470. https://doi.org/10.1039/c3tb21509a.CrossRefGoogle Scholar
- 24.Boontheekul T, Kong H-J, Mooney DJ. Controlling alginate gel degradation utilizing partial oxidation and bimodal molecular weight distribution. Biomaterials. 2005;26:2455–65. https://doi.org/10.1016/j.biomaterials.2004.06.044.CrossRefGoogle Scholar
- 34.Ivanovska J, Zehnder T, Lennert P, Sarker B, Boccaccini AR, Hartmann A. et al. Biofabrication of 3D alginate-based hydrogel for cancer research: comparison of cell spreading, viability, and adhesion characteristics of colorectal HCT116 tumor cells. Tissue Eng Part C Methods. 2016;22:708–15. https://doi.org/10.1089/ten.tec.2015.0452.CrossRefGoogle Scholar
- 37.Sarker B. advanced hydrogels concepts based on combinations of alginate, gelatin and bioactive glasses for tissue engineering. 2015. PhD thesis, Friedrich-Alexander University Erlangen-Nuremberg.Google Scholar
- 39.Cicha I, Goppelt-Struebe M, Muehlich S, Yilmaz A, Raaz D, Daniel WG. et al. Pharmacological inhibition of RhoA signaling prevents connective tissue growth factor induction in endothelial cells exposed to non-uniform shear stress. Atherosclerosis. 2008;196:136–45. https://doi.org/10.1016/j.atherosclerosis.2007.03.016.CrossRefGoogle Scholar
- 52.Sorrell JM, Baber MA, Caplan AI. A self-assembled fibroblast-endothelial cell co-culture system that supports in vitro vasculogenesis by both human umbilical vein endothelial cells and human dermal microvascular endothelial cells. Cells Tissues Organs. 2007;186:157–68. https://doi.org/10.1159/000106670.CrossRefGoogle Scholar
- 53.Asakawa N, Shimizu T, Tsuda Y, Sekiya S, Sasagawa T, Yamato M. et al. Pre-vascularization of in vitro three-dimensional tissues created by cell sheet engineering. Biomaterials. 2010;31:3903–9. https://doi.org/10.1016/j.biomaterials.2010.01.105.CrossRefGoogle Scholar