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Published Online: 6 August 2014

An Electrohydrodynamic Bioprinter for Alginate Hydrogels Containing Living Cells

Publication: Tissue Engineering Part C: Methods
Volume 21, Issue Number 2


In this work we present a bioprinting technique that exploits the electrohydrodynamic process to obtain a jet of liquid alginate beads containing cells. A printer is used to microfabricate hydrogels block by block following a bottom-up approach. Alginate beads constitute the building blocks of the microfabricated structures. The beads are placed at predefined position on a target substrate made of calcium-enriched gelatin, where they crosslink upon contact without the need of further postprocessing. The printed sample can be easily removed from the substrate at physiological temperature. Three-dimensional printing is accomplished by the deposition of multiple layers of hydrogel. We have investigated the parameters influencing the process, the compatibility of the printing procedure with cells, and their survival after printing.

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Mironov V., Visconti R.P., Kasyanov V., Forgacs G., Drake C.J., and Markwald R.R. Organ printing: tissue spheroids as building blocks. Biomaterials 30, 2164, 2009.
Bhadriraju K., and Chen C.S. Engineering cellular microenvironments to improve cell-based drug testing. Drug Discov Today 7, 612, 2002.
Elbert D.L. Bottom-up tissue engineering. Curr Opin Biotechnol 22, 674, 2011.
Khademhosseini A., and Langer R. Microengineered hydrogels for tissue engineering. Biomaterials 28, 5087, 2007.
Dimitrov D.S. Therapeutic proteins. Methods Mol Biol 899, 1, 2012.
De Vos P., Faas M.M., Strand B., and Calafiore R. Alginate-based microcapsules for immunoisolation of pancreatic islets. Biomaterials 27, 5603, 2006.
Murua A., Orive G., Hernández R.M., and Pedraz J.L. Xenogeneic transplantation of erythropoietin-secreting cells immobilized in microcapsules using transient immunosuppression. J Control Release 137, 174, 2009.
Chang P. The in vivo delivery of heterologous proteins by microencapsulated recombinant cells. Trends Biotechnol 17, 78, 1999.
Xu T., Jin J., Gregory C., Hickman J.J.J.J., and Boland T. Inkjet printing of viable mammalian cells. Biomaterials 26, 93, 2005.
Cui X., and Boland T. Human microvasculature fabrication using thermal inkjet printing technology. Biomaterials 30, 6221, 2009.
Barron J.A., Wu P., Ladouceur H.D., and Ringeisen B.R. Biological laser printing: a novel technique for creating heterogeneous 3-dimensional cell patterns. Biomed Microdevices 6, 139, 2004.
Ringeisen B.R., Kim H., Barron J.A., Krizman D.B., Chrisey D.B., Jackman S., et al. Laser printing of pluripotent embryonal carcinoma cells. Tissue Eng 10, 483, 2004.
Ringeisen B.R., Othon C.M., Barron J.A., Young D., and Spargo B.J. Jet-based methods to print living cells. Biotechnol J 1, 930, 2006.
Bock N., Dargaville T.R., and Woodruff M.A. Electrospraying of polymers with therapeutic molecules: State of the art. Prog Polym Sci 37, 1510, 2012.
Taylor G. Disintegration of water drops in electric field. Proc R Soc London 280, 383, 1964.
Park J.-U., Hardy M., Kang S.J., Barton K., Adair K., Mukhopadhyay D.K., et al. High-resolution electrohydrodynamic jet printing. Nat Mater 6, 782, 2007.
Moghadam H., Samimi M., Samimi A., and Khorram M. Electrospray modeling of highly viscous and non-Newtonian liquids. J Appl Polym Sci 118, 1288, 2010.
Gupta A., Seifalian A.M., Ahmad Z., Edirisinghe M.J., and Winslet M.C. Novel electrohydrodynamic printing of nanocomposite biopolymer scaffolds. J Bioact Compat Polym 22, 265, 2007.
Mongkoldhumrongkul N., Best S., Aarons E., and Jayasinghe S.N. Bio-electrospraying whole human blood: analysing cellular viability at a molecular level. J Tissue Eng Regen Med 3, 562, 2009.
Clarke J.D.W., and Jayasinghe S.N. Bio-electrosprayed multicellular zebrafish embryos are viable and develop normally. Biomed Mater 3, 011001, 2008.
Xie J., and Wang C.-H. Electrospray in the dripping mode for cell microencapsulation. J Colloid Interface Sci 312, 247, 2007.
Murua A., Portero A., Orive G., Hernández R.M., de Castro M., and Pedraz J.L. Cell microencapsulation technology: towards clinical application. J Control Release 132, 76, 2008.
Peirone M., Ross C.J., Hortelano G., Brash J.L., and Chang P.L. Encapsulation of various recombinant mammalian cell types in different alginate microcapsules. J Biomed Mater Res 42, 587, 1998.
Duvivier-Kali V.F., Omer A., Parent R.J., O'Neil J.J., and Weir G.C. Complete protection of islets against allorejection and autoimmunity by a simple barium-alginate membrane. Diabetes 50, 1698, 2001.
Rowley J.A., Madlambayan G., and Mooney D.J. Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials 20, 45, 1999.
Lee K.Y., and Mooney D.J. Alginate: properties and biomedical applications. Prog Polym Sci 37, 106, 2012.
Yang J.-S., Xie Y.-J., and He W. Research progress on chemical modification of alginate: a review. Carbohydr Polym 84, 33, 2010.
Siti-Ismail N., Bishop A.E., Polak J.M., and Mantalaris A. The benefit of human embryonic stem cell encapsulation for prolonged feeder-free maintenance. Biomaterials 29, 3946, 2008.
Klouda L., and Mikos A.G. Thermoresponsive hydrogels in biomedical applications. Eur J Pharm Biopharm 68, 34, 2008.
Pataky K., Braschler T., Negro A., Renaud P., Lutolf M.P., and Brugger J. Microdrop printing of hydrogel bioinks into 3D tissue-like geometries. Adv Mater 24, 391, 2011.
Teramura Y., and Iwata H. Bioartificial pancreas microencapsulation and conformal coating of islet of Langerhans. Adv Drug Deliv Rev 62, 827, 2010.
Jacobs V., Anandjiwala R.D., and Maaza M. The influence of electrospinning parameters on the structural morphology and diameter of electrospun nanofibers. J Appl Polym Sci 115, 3130, 2010.
Manojlovic V., Djonlagic J., Obradovic B., Nedovic V., and Bugarski B. Investigations of cell immobilization in alginate: rheological and electrostatic extrusion studies. J Chem Technol Biotechnol 81, 505, 2006.
Hayati I., Bailey A., and Tadros T. Investigations into the mechanisms of electrohydrodynamic spraying of liquids: I. Effect of electric field and the environment on pendant drops and factors affecting the. J Colloid Interface Sci 117, 205, 1987.
Cloupeau M., and Prunet-Foch B. Electrohydrodynamic spraying functioning modes: a critical review. J Aerosol Sci 25, 1021, 1994.
Jun Y., Kim M.J., Hwang Y.H., Jeon E.A., Kang A.R., Lee S.-H., et al. Microfluidics-generated pancreatic islet microfibers for enhanced immunoprotection. Biomaterials 34, 8122, 2013.
Choi H.K., Park J.-U., Park O.O., Ferreira P.M., Georgiadis J.G., and Rogers J.a. Scaling laws for jet pulsations associated with high-resolution electrohydrodynamic printing. Appl Phys Lett 92, 123109, 2008.
Gasperini L., Maniglio D., and Migliaresi C. Microencapsulation of cells in alginate through an electrohydrodynamic process. J Bioact Compat Polym 28, 413, 2013.
Kuo C.K., and Ma P.X. Ionically crosslinked alginate hydrogels as scaffolds for tissue engineering: part 1. Structure, gelation rate and mechanical properties. Biomaterials 22, 511, 2001.
Blandino A.N.A., Macias M., and Canter D. Formation of Calcium Alginate Gel Capsules: Influence of Sodium Alginate and CaC12 Concentration on Gelation Kinetics 88, 686, 1999.
Ma H.-L., Hung S.-C., Lin S.-Y., Chen Y.-L., and Lo W.-H. Chondrogenesis of human mesenchymal stem cells encapsulated in alginate beads. J Biomed Mater Res A 64, 273, 2003.
Bouhadir K.H., Lee K.Y., Alsberg E., Damm K.L., Anderson K.W., and Mooney D.J. Degradation of partially oxidized alginate and its potential application for tissue engineering. Biotechnol Prog 17, 945, 2001.
Gomez C., Rinaudo M., and Villar M. Oxidation of sodium alginate and characterization of the oxidized derivatives. Carbohydr Polym 67, 296, 2007.
Yu J., Gu Y., Du K.T., Mihardja S., Sievers R.E., and Lee R.J. The effect of injected RGD modified alginate on angiogenesis and left ventricular function in a chronic rat infarct model. Biomaterials 30, 751, 2009.
Tasoglu S., and Demirci U. Bioprinting for stem cell research. Trends Biotechnol 31, 10, 2013.
Boland T., Xu T., Damon B., and Cui X. Application of inkjet printing to tissue engineering. Biotechnol J 1, 910, 2006.
Mironov V., Kasyanov V., and Markwald R.R. Organ printing: from bioprinter to organ biofabrication line. Curr Opin Biotechnol 22, 667, 2011.
Derby B. Printing and prototyping of tissues and scaffolds. Science 338, 921, 2012.
Jayasinghe S.N., Qureshi A.N., and Eagles P.A.M. Electrohydrodynamic jet processing: an advanced electric-field-driven jetting phenomenon for processing living cells. Small 2, 216, 2006.
Schiele N.R., Corr D.T., Huang Y., Raof N.A., Xie Y., and Chrisey D.B. Laser-based direct-write techniques for cell printing. Biofabrication 2, 032001, 2010.
Elisseeff J., McIntosh W., Anseth K., Riley S., Ragan P., and Langer R. Photoencapsulation of chondrocytes in poly(ethylene oxide)-based semi-interpenetrating networks. J Biomed Mater Res 51, 164, 2000.
Yang F., Williams C.G., Wang D.-A., Lee H., Manson P.N., and Elisseeff J. The effect of incorporating RGD adhesive peptide in polyethylene glycol diacrylate hydrogel on osteogenesis of bone marrow stromal cells. Biomaterials 26, 5991, 2005.
Morota K., Matsumoto H., Mizukoshi T., Konosu Y., Minagawa M., Tanioka A., et al. Poly(ethylene oxide) thin films produced by electrospray deposition: morphology control and additive effects of alcohols on nanostructure. J Colloid Interface Sci 279, 484, 2004.
Cruise G.M., Hegre O.D., Scharp D.S., and Hubbell J.a. A sensitivity study of the key parameters in the interfacial photopolymerization of poly(ethylene glycol) diacrylate upon porcine islets. Biotechnol Bioeng 57, 655, 1998.
Francisco H., Yellen B.B., Halverson D.S., Friedman G., and Gallo G. Regulation of axon guidance and extension by three-dimensional constraints. Biomaterials 28, 3398, 2007.
Schnell E., Klinkhammer K., Balzer S., Brook G., Klee D., Dalton P., et al. Guidance of glial cell migration and axonal growth on electrospun nanofibers of poly-ɛ-caprolactone and a collagen/poly-ɛ-caprolactone blend. Biomaterials 28, 3012, 2007.

Information & Authors


Published In

cover image Tissue Engineering Part C: Methods
Tissue Engineering Part C: Methods
Volume 21Issue Number 2February 2015
Pages: 123 - 132
PubMed: 24903714


Published in print: February 2015
Published ahead of print: 8 August 2014
Published online: 6 August 2014
Published ahead of production: 6 June 2014
Accepted: 30 May 2014
Received: 7 March 2014


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Luca Gasperini
Department of Industrial Engineering, Biotech Research Center, University of Trento, Trento, Italy.
European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Trento, Italy.
Devid Maniglio
Department of Industrial Engineering, Biotech Research Center, University of Trento, Trento, Italy.
European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Trento, Italy.
INSTM—Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali, Firenze, Italy.
Antonella Motta
Department of Industrial Engineering, Biotech Research Center, University of Trento, Trento, Italy.
European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Trento, Italy.
INSTM—Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali, Firenze, Italy.
Claudio Migliaresi
Department of Industrial Engineering, Biotech Research Center, University of Trento, Trento, Italy.
European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Trento, Italy.
INSTM—Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali, Firenze, Italy.


Address correspondence to:Claudio Migliaresi, PhDDepartment of Industrial EngineeringUniversity of TrentoVia Sommarive 938123 TrentoItaly
E-mail: [email protected]

Disclosure Statement

No competing financial interests exist.

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