Guidelines for standardization of bioprinting: a systematic study of process parameters and their effect on bioprinted structures

Matti Kesti 1 , Philipp Fisch 1 , Marco Pensalfini 2 , 3 , Edoardo Mazza 2 , 3  and Marcy Zenobi-Wong 4
  • 1 Institute for Biomechanics, Switzerland
  • 2 Institute for Mechanical Systems, Switzerland
  • 3 Swiss Federal Laboratories for Materials Science and Technology, Switzerland
  • 4 Institute for Biomechanics, Switzerland
Matti Kesti, Philipp Fisch, Marco Pensalfini
  • Institute for Mechanical Systems, Switzerland
  • Swiss Federal Laboratories for Materials Science and Technology, Switzerland
  • Search for other articles:
  • degruyter.comGoogle Scholar
, Edoardo Mazza
  • Institute for Mechanical Systems, Switzerland
  • Swiss Federal Laboratories for Materials Science and Technology, Switzerland
  • Search for other articles:
  • degruyter.comGoogle Scholar
and Marcy Zenobi-Wong

Abstract

Biofabrication techniques including three-dimensional bioprinting could be used one day to fabricate living, patient-specific tissues and organs for use in regenerative medicine. Compared to traditional casting and molding methods, bioprinted structures can be much more complex, containing for example multiple materials and cell types in controlled spatial arrangement, engineered porosity, reinforcement structures and gradients in mechanical properties. With this complexity and increased function, however, comes the necessity to develop guidelines to standardize the bioprinting process, so printed grafts can safely enter the clinics. The bioink material must firstly fulfil requirements for biocompatibility and flow. Secondly, it is important to understand how process parameters affect the final mechanical properties of the printed graft. Using a gellan-alginate physically crosslinked bioink as an example, we show shear thinning and shear recovery properties which allow good printing resolution. Printed tensile specimens were used to systematically assess effect of line spacing, printing direction and crosslinking conditions. This standardized testing allowed direct comparison between this bioink and three commercially-available products. Bioprinting is a promising, yet complex fabrication method whose outcome is sensitive to a range of process parameters. This study provides the foundation for highly needed best practice guidelines for reproducible and safe bioprinted grafts.

  • 1.

    Chhaya PM, Poh SP, Balmayor RE, van Griensven M, Schantz J-T, Hutmacher WD. Additive manufacturing in biomedical sciences and the need for definitions and norms. Expert Rev Med Devices. 2015;12:537–43.

  • 2.

    Malda J, Visser J, Melchels PF, Jüngst T, Hennink EW, Dhert JW, et al. 25th anniversary article: engineering hydrogels for biofabrication. Adv Mat. 2013;25:5011–28.

  • 3.

    Billiet T, Van Gasse B, Gevaert E, Cornelissen M, Martins CJ, Dubruel P. Quantitative contrasts in the photopolymerization of acrylamide and methacrylamide-functionalized gelatin hydrogel building blocks. Macromol Biosci. 2013;13:1531–45.

  • 4.

    Aguado B, Mulyasasmita W, Su J, Lampe K, Heilshorn S. Improving viability of stem cells during syringe needle flow through the design of hydrogel cell carriers. Tissue Eng Part A. 2012;18:806–15.

  • 5.

    Billiet T, Gevaert E, De Schryver T, Cornelissen M, Dubruel P. The 3D printing of gelatin methacrylamide cell-laden tissue-engineered constructs with high cell viability. Biomaterials. 2014;35:49–62.

  • 6.

    Khalil S, Sun W. Bioprinting endothelial cells with alginate for 3D tissue constructs. J Biomec Eng. 2009;131:1–8.

  • 7.

    Tirella AO, Vozzi G, Ahluwalia A. A phase diagram for microfabrication of geometrically controlled hydrogel scaffolds. Biofabrication. 2009;1:1–12.

  • 8.

    Mazza E, Ehret EA. Mechanical biocompatibility of highly deformable biomedical materials. J Mech Behav Biomed Mat. 2015;48:100–24.

  • 9.

    Bakarich S, Balding P, Gorkin R, Spinks GM, In het Panhuis, M. Printed ionic-covalent entanglement hydrogels from carrageenan and an epoxy amine. RSC Adv. 2014;4:38088–92.

  • 10.

    Bakarich S, Gorkin R, In Het Panhuis M, Spinks GM. Three-dimensional printing fiber reinforced hydrogel composites. ACS Appl Mater Interfaces. 2014;6:15998–6006.

  • 11.

    Compton GB, Lewis AJ. 3D-Printing of lightweight cellular composites. Adv Mat. 2014;26:5930–5.

  • 12.

    Cui J, Lackey M, Madkour EA, Saffer ME, Griffin MD, Bhatia RS, et al. Synthetically simple, highly resilient hydrogels. Biomacromol. 2012;13:584–588.

  • 13.

    Hong S, Sycks D, Chan H, Lin S, Lopez PG, Guilak F, et al. 3D printing of highly stretchable and tough hydrogels into complex, cellularized structures. Adv Mat. 2015;27:4035–40.

  • 14.

    Kesti M, Eberhardt C, Pagliccia G, Kenkel D, Grande D, Boss A, et al. Bioprinting complex cartilaginous structures with clinically compliant biomaterials. Adv Func Mat. 2015;25:7406–17.

  • 15.

    McKee TC, Last AJ, Russell P, Murphy JC. Indentation versus tensile measurements of young’s modulus for soft biological tissues. Tissue Eng Part B. 2011;17:155–64.

  • 16.

    Mueller J, Shea K, Daraio C. Mechanical properties of parts fabricated with inkjet 3D printing through efficient experimental design. Mater Des. 2015;86:902–12.

  • 17.

    Wei J, Wang J, Su S, Wang S, Qiu J, Zhang Z, et al. 3D printing of an extremely tough hydrogel. Roy Soc Chem Adv. 2015;5:81324–9.

  • 18.

    Hoch E, Schuh C, Hirth T, Tovar ME, Borchers K. Stiff gelatin hydrogels can be photo-chemically synthesized from low viscous gelatin solutions using molecularly functionalized gelatin with a high degree of methacrylation. J Mat Sci Mater Med. 2012;23:2607–17.

  • 19.

    Schuurman W, Levett P, Pot WM, Van Weeren RP, Dhert AJ, Hutmacher WD, et al. Gelatin-methacrylamide hydrogels as potential biomaterials for fabrication of tissue-engineered cartilage constructs. Macromol Biosci. 2013;13:551–61.

  • 20.

    Rimann M, Bono E, Annaheim H, Bleisch M, Graf-Hausner U. Standardized 3D bioprinting of soft tissue models with human primary cells. J Lab Autom. 2015;1–14.

  • 21.

    Markstedt K, Mantas A, Tournier I, Martinez Avila H, Hägg D, Gatenholm P. 3D bioprinting human chondrocytes with nanocellulose–alginate bioink for cartilage tissue engineering applications. Biomacromol. 2015;16:1489–96.

  • 22.

    Grasdalen H, Smidsrød O. Gelation of Gellan Gum. Carbohyd Polym. 1987;7:371–93.

  • 23.

    Mørch AY, Donati I, Strand LB, Skjak-Braek G. Effect of Ca2+, Ba2+, and Sr2+ on alginate microbeads. Biomacromol. 2006;7:1471–80.

  • 24.

    Bryant JS, Nuttelman RC, Anseth SK. Cytocompatibility of UV and visible light photoinitiating systems on cultured NIH/3T3 fibroblasts in vitro. J Biomat Sci Polymer Edn. 2000;11:439–57.

  • 25.

    Williams GC, Malik NA, Kim KT, Manson NP, Elisseeff HJ. Variable cytocompatibility of six cell lines with photoinitiators used for polymerizing hydrogels and cell encapsulation. Biomat. 2005;26:1211–8.

  • 26.

    Hopf R, Bernardi L, Menze J, Zündel M, Mazza E. Experimental and theoretical analyses of the age-dependent large-strain behavior of Sylgard 184 (10:1) silicone elastomer. J Mech Behavior Biomed Mat. 2016;60:425–437.

Purchase article
Get instant unlimited access to the article.
Log in
Already have access? Please log in.


or
Log in with your institution

Journal + Issues

Search