3D Bioprinting Technologies for Cellular Engineering


Bioprinting encompasses the use of additive manufacturing methods for the purpose of creating cellular constructs of varying complexity into prescribed geometrical forms (i.e. individual cells, cell agglomerates, tissues, and organs). Collectively, these methods offer many advantages over scaffold-based fabrication, including the ability to pattern complex cellular constructs on relevant length scales, the ability to tailor and modulate the extracellular environment with high precision, a means to study cell differentiation and proliferation under conditions that mimic natural biological environments, and a means to fabricate 3d tissue constructs of geometrical complexity approaching that of biological systems. Unlike industrial additive manufacturing, however, bioprinting faces additional challenges that deal with cell sensitivity and viability, the need for precise spatial and chemical tuning of the extra cellular environment, and more generally the creation of functional constructs that approximate biological tissue. In this chapter, we discuss how these challenges are being met by various bioprinting approaches, with a focus on the underlying mechanical and biological principles.


Additive Manufacturing Inkjet Printing Tissue Construct Laser Direct Writing Multicellular Aggregate 


  1. 1.
  2. 2.
    Global 3D Bioprinting Market (2014–2018), TechNavio—Infiniti Research Ltd., 2014Google Scholar
  3. 3.
    Oberpenning F, Meng J, Yoo JJ, Atala A (1999) De novo reconstitution of a functional mammalian urinary bladder by tissue engineering. Nat Biotechnol 17(2):149–155CrossRefGoogle Scholar
  4. 4.
    Atala A, Bauer SB, Soker S, Yoo JJ, Retik AB (2006) Tissue-engineered autologous bladders for patients needing cystoplasty. Lancet 367(9518):1241–1246CrossRefGoogle Scholar
  5. 5.
    Reiffel AJ, Kafka C, Hernandez KA, Popa S, Perez JL, Zhou S, Spector JA (2013) High-fidelity tissue engineering of patient-specific auricles for reconstruction of pediatric microtia and other auricular deformities. PLoS One 8(2):e56506CrossRefGoogle Scholar
  6. 6.
    Lanza R, Langer R, Vacanti JP (eds) (2011) Principles of tissue engineering. Academic Press, New YorkGoogle Scholar
  7. 7.
    Marga F, Jakab K, Khatiwala C, Shephard B, Dorfman S, Forgacs G (2012) Organ printing: a novel tissue engineering paradigm. In: 5th European conference of the international federation for medical and biological engineering, Springer, Berlin, pp 27–30Google Scholar
  8. 8.
    Griffith CK, Miller C, Sainson R, Calvert JW, Jeon NL, Hughes CW, George SC (2005) Tissue Eng 11:257CrossRefGoogle Scholar
  9. 9.
    Lodish H, Arnold B, Matsudaira P, Kaiser C, Krieger M, Scott M, Zipursky SL, Darnell J (2000) Molecular cell biology. W.H. Freeman and Company, New YorkGoogle Scholar
  10. 10.
    Jakab K, Norotte C, Marga F, Murphy K, Vunjak-Novakovic G, Forgacs G (2010) Tissue engineering by self-assembly and bio-printing of living cells. Biofabrication 2(2):022001CrossRefGoogle Scholar
  11. 11.
    Mironov V, Visconti RP, Kasyanov V, Forgacs G, Drake CJ, Markwald RR (2009) Organ printing: tissue spheroids as building blocks. Biomaterials 30(12):2164–2174CrossRefGoogle Scholar
  12. 12.
    Jakab K, Damon B, Neagu A, Kachurin A, Forgacs G (2006) Three-dimensional tissue constructs built by bioprinting. Biorheology 43:509–513Google Scholar
  13. 13.
    Norotte C, Marga FS, Niklason LE, Forgacs G (2009) Scaffold-free vascular tissue engineering using bioprinting. Biomaterials 30:5910–5917CrossRefGoogle Scholar
  14. 14.
    Willie W, DeConinck A, Lewis JA (2011) Omnidirectional printing of 3D microvascular networks. Adv Mater 23(24):H178–H183CrossRefGoogle Scholar
  15. 15.
    Kolesky DB et al (2014) 3D bioprinting of vascularized, heterogeneous cell‐laden tissue constructs. Adv Mater 26(19):3124–3130CrossRefGoogle Scholar
  16. 16.
    Cohen DL et al (2010) Additive manufacturing for in situ repair of osteochondral defects. Biofabrication 2(3):035004Google Scholar
  17. 17.
    Klebe RJ (1988) Cytoscribing: a method for micropositioning cells and the construction of two-and three-dimensional synthetic tissues. Exp Cell Res 179(2):362–373CrossRefGoogle Scholar
  18. 18.
    Klebe RJ et al (1994) Cytoscription: computer controlled micropositioning of cell adhesion proteins and cells. J Tissue Cult Methods 16(3–4):189–192CrossRefGoogle Scholar
  19. 19.
    Hutchings IM, Martin GD (eds) (2012) Inkjet technology for digital fabrication. Wiley, New YorkGoogle Scholar
  20. 20.
    Hon KKB, Li L, Hutchings IM (2008) Direct writing technology – advances and developments. CIRP Ann Manuf Technol 57(2):601–620CrossRefGoogle Scholar
  21. 21.
    Demirci U, Montesano G (2007) Single cell epitaxy by acoustic picoliter droplets. Lab Chip 7:1139–1145CrossRefGoogle Scholar
  22. 22.
    Xu T et al (2005) Inkjet printing of viable mammalian cells. Biomaterials 26(1):93–99Google Scholar
  23. 23.
    Nakamura M et al (2005) Biocompatible inkjet printing technique for designed seeding of individual living cells. Tissue Eng 11(11–12):1658–1666CrossRefGoogle Scholar
  24. 24.
    Saunders RE, Gough JE, Derby B (2008) Biomaterials 29:193–203CrossRefGoogle Scholar
  25. 25.
    Xu T, Gregory CA, Molnar P, Cui X, Jalota S, Bhaduri SB, Boland T (2006) Biomaterials 27:3580–3588Google Scholar
  26. 26.
    Reis N, Ainsley C, Derby B (2005) Ink: jet delivery of particle suspensions by piezoelectric droplet ejectors. J Appl Phys 97(9):094903Google Scholar
  27. 27.
    Nishioka GM, Markey AA, Holloway CK (2004) J Am Chem Soc 126:16320–16321CrossRefGoogle Scholar
  28. 28.
    Di Risio S, Yan N (2007) Macromol. Rapid Commun 28:1934–1940CrossRefGoogle Scholar
  29. 29.
    Ilkhanizadeh S, Teixeira AI, Hermanson O (2007) Biomaterials 28:3936–3943CrossRefGoogle Scholar
  30. 30.
    Phillippi JA, Miller E, Weiss L, Huard J, Waggoner A, Campbell P (2008) Stem Cells 26:127–134CrossRefGoogle Scholar
  31. 31.
    Tao X et al (2013) Complex heterogeneous tissue constructs containing multiple cell types prepared by inkjet printing technology. Biomaterials 34(1):130–139CrossRefGoogle Scholar
  32. 32.
    US Patent No. 4,575,330Google Scholar
  33. 33.
    Elomaa L, Teixeira S, Hakala R, Korhonen H, Grijpma DW, Seppala JV (2011) Preparation of poly(epsilon-caprolactone)-based tissue engineering scaffolds by stereolithography. Acta Biomater 7:3850–3856CrossRefGoogle Scholar
  34. 34.
    Lee JW, Lan PX, Kim B, Lim G, Cho DW (2008) Fabrication and characteristic analysis of a poly(propylene fumarate) scaffold using micro-stereolithography technology. J Biomed Mater Res B 87B:1–9CrossRefGoogle Scholar
  35. 35.
    Seck TM, Melchels FPW, Feijen J, Grijpma DW (2010) Designed biodegradable hydrogel structures prepared by stereolithography using poly(ethylene glycol)/poly(d, l-lactide)-based resins. J Control Release 148:34–41CrossRefGoogle Scholar
  36. 36.
    Zhang X, Jiang XN, Sun C (1999) Micro-stereolithography of polymeric and ceramic microstructures. Sensors Actuators A Phys 77(2):149–156CrossRefGoogle Scholar
  37. 37.
    Seitz H et al (2005) Three‐dimensional printing of porous ceramic scaffolds for bone tissue engineering. J Biomed Mater Res B Appl Biomater 74(2):782–788CrossRefGoogle Scholar
  38. 38.
    Sabree I (2014) Fabrication of bioactive glass scaffolds by stereolithography for bone tissue engineering. [Thesis]. The University of Manchester, Manchester, UKGoogle Scholar
  39. 39.
    Elomaa L et al (2013) Porous 3D modeled scaffolds of bioactive glass and photocrosslinkable poly (ε-caprolactone) by stereolithography. Compos Sci Technol 74:99–106CrossRefGoogle Scholar
  40. 40.
    Dhariwala B, Hunt E, Boland T (2004) Rapid prototyping of tissue-engineering constructs, using photopolymerizable hydrogels and stereolithography. Tissue Eng 10(9–10):1316–1322CrossRefGoogle Scholar
  41. 41.
    Arcaute K, Mann BK, Wicker RB (2006) Stereolithography of three-dimensional bioactive poly(ethylene glycol) constructs with encapsulated cells. Ann Biomed Eng 34:1429–1441CrossRefGoogle Scholar
  42. 42.
    Chan V, Zorlutuna P, Jeong JH, Kong H, Bashir R (2010) Three-dimensional photopatterning of hydrogels using stereolithography for long-term cell encapsulation. Lab Chip 10:2062–2070CrossRefGoogle Scholar
  43. 43.
    Zorlutuna P, Jeong JH, Kong H, Bashir R (2011) Stereolithography-based hydrogel microenvironments to examine cellular interactions. Adv Funct Mater 21:3642–3651CrossRefGoogle Scholar
  44. 44.
    Choi J-W et al (2009) Fabrication of 3D biocompatible/biodegradable micro-scaffolds using dynamic mask projection microstereolithography. J Mater Process Technol 209(15):5494–5503CrossRefGoogle Scholar
  45. 45.
    Cheng YL, Lee ML (2009) Development of dynamic masking rapid prototyping system for application in tissue engineering. Rapid Prototyp J 15(1):29–41CrossRefGoogle Scholar
  46. 46.
    Han L-H et al (2008) Projection microfabrication of three-dimensional scaffolds for tissue engineering. J Manuf Sci Eng 130(2):021005CrossRefGoogle Scholar
  47. 47.
    Han L-H et al (2010) Fabrication of three-dimensional scaffolds for heterogeneous tissue engineering. Biomed Microdevices 12(4):721–725CrossRefGoogle Scholar
  48. 48.
    Gauvin R et al (2012) Microfabrication of complex porous tissue engineering scaffolds using 3D projection stereolithography. Biomaterials 33(15):3824–3834Google Scholar
  49. 49.
    Maruo S, Nakamura O, Kawata S (1997) Three-dimensional microfabrication with two-photon-absorbed photopolymerization. Opt Lett 22:132–134CrossRefGoogle Scholar
  50. 50.
    Maruo S, Kawata S (1998) Two-photon-absorbed near-infrared photopolymerization for three-dimensional microfabrication. J Microelectromech Syst 7:411–415CrossRefGoogle Scholar
  51. 51.
    Pitts JD, Campagnola PJ, Epling GA, Goodman SL (2000) Submicron multiphoton free-form fabrication of proteins and polymers: studies of reaction efficiencies and applications in sustained release. Macromolecules 33:1514–1523CrossRefGoogle Scholar
  52. 52.
    Kaehr B, Shear JB (2007) Mask-directed multiphoton lithography. J Am Chem Soc 129:1904–1905CrossRefGoogle Scholar
  53. 53.
    Nielson R, Koehr B, Shear JB (2009) Microreplication and design of biological architectures using dynamic-mask multiphoton lithography. Small 5:120–125CrossRefGoogle Scholar
  54. 54.
    Luo Y, Shoichet MS (2004) A photolabile hydrogel for guided three-dimensional cell growth and migration. Nat Mater 3:249–253CrossRefGoogle Scholar
  55. 55.
    Aizawa Y, Leipzig N, Zahir T, Shoichet M (2008) The effect of immobilized platelet derived growth factor aa on neural stem/progenitor cell differentiation on cell-adhesive hydrogels. Biomaterials 29:4676–4683CrossRefGoogle Scholar
  56. 56.
    Leipzig ND, Wylie RG, Kim H, Shoichet MS (2011) Differentiation of neural stem cells in three-dimensional growth factor-immobilized chitosan hydrogel scaffolds. Biomaterials 32:57–64CrossRefGoogle Scholar
  57. 57.
    Wosnick JH, Shoichet MS (2008) Three-dimensional chemical patterning of transparent hydrogels. Chem Mater 20:55–60CrossRefGoogle Scholar
  58. 58.
    Wylie RG, Shoichet MS (2011) Three-dimensional spatial patterning of proteins in hydrogels. Biomacromolecules 12:3789–3796CrossRefGoogle Scholar
  59. 59.
    Wylie RG, Ahsan S, Aizawa Y, Maxwell KL, Morshead CM, Shoichet MS (2011) Spatially controlled simultaneous patterning of multiple growth factors in three-dimensional hydrogels. Nat Mater 10:799–806CrossRefGoogle Scholar
  60. 60.
    Ovsianikov A et al (2010) Laser printing of cells into 3D scaffolds. Biofabrication 2(1):014104CrossRefGoogle Scholar
  61. 61.
    Ovsianikov A et al (2011) Three-dimensional laser micro-and nano-structuring of acrylated poly (ethylene glycol) materials and evaluation of their cytoxicity for tissue engineering applications. Acta Biomater 7(3):967–974CrossRefGoogle Scholar
  62. 62.
    Li Y, Maynor BW, Liu J (2001) J Am Chem Soc 123:2105CrossRefGoogle Scholar
  63. 63.
    Amro NA, Xu S, Liu GY (2000) Langmuir 16:3006CrossRefGoogle Scholar
  64. 64.
    Wang XF, Ryu KS, Bullen DA, Zou J, Zhang H, Mirkin CA, Liu C (2003) Langmuir 19:8951CrossRefGoogle Scholar
  65. 65.
    Huo FW, Zheng ZJ, Zheng GF, Giam LR, Zhang H, Mirkin CA (2008) Science 321:1658CrossRefGoogle Scholar
  66. 66.
    Kim KH, Moldovan N, Espinosa HD (2005) A nanofountain probe with sub-100 nm molecular writing resolution. Small 1(6):632–635CrossRefGoogle Scholar
  67. 67.
    Bohandy J, Kim BF, Adrian FJ (1986) Metal deposition from a supported metal film using an excimer laser. J Appl Phys 60(4):1538–1539CrossRefGoogle Scholar
  68. 68.
    Barron JA, Wu P, Ladouceur HD, Ringeisen BR (2004) Biological laser printing: a novel technique for creating heterogeneous 3-dimensional cell patterns. Biomed Microdevices 6(2):139–147CrossRefGoogle Scholar
  69. 69.
    Barron JA, Spargo BJ, Ringeisen BR (2004) Biological laser printing of three dimensional cellular structures. Appl Phys A 79(4–6):1027–1030Google Scholar
  70. 70.
    Koch L et al (2009) Laser printing of skin cells and human stem cells. Tissue Eng C Methods 16(5):847–854CrossRefGoogle Scholar
  71. 71.
    Nahmias Y et al (2005) Laser‐guided direct writing for three‐dimensional tissue engineering. Biotechnol Bioeng 92(2):129–136CrossRefGoogle Scholar
  72. 72.
    Othon CM et al (2008) Single-cell printing to form three-dimensional lines of olfactory ensheathing cells. Biomed Mater 3(3):034101CrossRefGoogle Scholar
  73. 73.
    Ashkin A (1970) Atomic-beam deflection by resonance-radiation pressure. Phys Rev Lett 25(19):1321CrossRefGoogle Scholar
  74. 74.
    Odde DJ, Renn MJ (2000) Laser‐guided direct writing of living cells. Biotechnol Bioeng 67(3):312–318CrossRefGoogle Scholar
  75. 75.
    Murphy SV, Atala A (2014) 3D bioprinting of tissues and organs. Nat Biotechnol 32(8):773–785CrossRefGoogle Scholar
  76. 76.
    Wallace J et al (2014) Validating continuous digital light processing (cDLP) additive manufacturing accuracy and tissue engineering utility of a dye-initiator package. Biofabrication 6(1):015003CrossRefGoogle Scholar
  77. 77.
    Piner RD et al (1999) “Dip-pen” nanolithography. Science 283(5402):661–663CrossRefGoogle Scholar
  78. 78.
    Ringeisen BR et al (eds) (2010) Cell and organ printing. Springer Science and Business Media BV, New YorkGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2016

Authors and Affiliations

  1. 1.Cornell UniversityIthacaUSA

Personalised recommendations