Progress in Additive Manufacturing

, Volume 3, Issue 4, pp 245–255 | Cite as

Multi-layer cryolithography for additive manufacturing

  • Bartłomiej Zawada
  • Gideon UkpaiEmail author
  • Matthew J. Powell-Palm
  • Boris Rubinsky
Full Research Article


A new technique is introduced which addresses the need for faster additive manufacturing methods for tissue scaffolds and frozen foods in large-scale industrial applications, inspired by print lithography. It is particularly relevant to biological matter, which is composed mostly of water. Instead of point-by-point printing in three dimensions (3D) with 3D printers, multiple single 2D layers can be assembled or printed separately, in parallel, on areas coated with hydrophilic materials to bind water-based compounds and hydrophobic materials to reject water-based compounds and bind hydrophobic molecules. This technique keeps the layers attached to the surface, opposing gravity, and thereby facilitating the transport and the assembly of the 2D layers, regardless of the direction of the surface relative to gravity. The individual layers are deposited one on top of the other and linked by chemical cross-linking and freezing to generate a 3D structure. Examples show how complex and large hydrogel-based structures can be manufactured by multi-layer cryolithography from fusion and freezing of 2D layers. Applications involve tissue engineering and food engineering, with particular emphasis on the ability to assemble a biological object, while every volume is frozen under optimal conditions during the assembly. Scanning electron microscopy demonstrates the ability to control and produce uniform microstructures in the 3D objects produced by cryolithography.


Cryolithography Bioprinting Tissue engineering Food printing 3D printing 3D print lithography 


Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.


  1. 1.
    Weaver P (1964) The technique of lithography. Reinhold Pub., LondonGoogle Scholar
  2. 2.
    Gibson I, Rosen DW, Stucker B (2010) Additive manufacturing technologies: rapid prototyping to direct digital manufacturing. Springer, BerlinCrossRefGoogle Scholar
  3. 3.
    Ozbolat IT, Yu Y (2013) Bioprinting toward organ fabrication: challenges and future trends. IEEE Trans Biomed Eng 60:691–699. CrossRefGoogle Scholar
  4. 4.
    Chang TT, Zhou VX, Rubinsky B (2017) Using non-thermal irreversible electroporation to create an in vivo niche for exogenous cell engraftment. Biotechniques 62:229–231. CrossRefGoogle Scholar
  5. 5.
    Lee VK, Dai G (2017) Printing of three-dimensional tissue analogs for regenerative medicine. Ann Biomed Eng 45:115–131CrossRefGoogle Scholar
  6. 6.
    Skardal A, Atala A (2015) Biomaterials for integration with 3-D bioprinting. Ann Biomed Eng 43:730–746. CrossRefGoogle Scholar
  7. 7.
    Murphy SV, Skardal A, Atala A (2013) Evaluation of hydrogels for bio-printing applications. J Biomed Mater Res Part A 101:272–284. CrossRefGoogle Scholar
  8. 8.
    Yannas IV, Burke JF (1980) Design of an artificial skin. 1. Basic design principles. J Biomed Mater Res 14:65–81. CrossRefGoogle Scholar
  9. 9.
    Shapiro L, Cohen S (1997) Novel alginate sponges for cell culture and transplantation. Biomaterials 18:583–590. CrossRefGoogle Scholar
  10. 10.
    Rubinsky B (1983) Solidification processes in saline solutions. J Cryst Growth 62:513–522. CrossRefGoogle Scholar
  11. 11.
    Tsai HL, Rubinsky B (1984) A numerical study using “front tracking” finite elements on the morphological stability of a planar interface during transient solidification processes. J Cryst Growth 69:29–46. CrossRefGoogle Scholar
  12. 12.
    Rubinsky B, Ikeda M (1985) A cryosmicroscope using directional solidification for the controlled freezing of biological matter. Cryobiology 22:55–68CrossRefGoogle Scholar
  13. 13.
    Tsai HL, Rubinsky B (1984) A “front tracking” finite element study on change of phase interface stability during solidification processes in solutions. J Cryst Growth 70:56–63. CrossRefGoogle Scholar
  14. 14.
    Rubinsky B, Lee CY, Bastacky J, Hayes TL (1987) The mechanism of freezing in biological tissue—the liver. Cryo Lett 8:370–381Google Scholar
  15. 15.
    Zmora S, Glicklis R, Cohen S (2002) Tailoring the pore architecture in 3-D alginate scaffolds by controlling the freezing regime during fabrication. Biomaterials 23:4087–4094. CrossRefGoogle Scholar
  16. 16.
    Preciado JA, Skandakumaran P, Cohen S, Rubinsky B (2003) Utilization of directional freezing for the construction of tissue engineering scaffolds. Heat Transf 4:439–442Google Scholar
  17. 17.
    Xiong Z, Yan Y, Wang S et al (2002) Fabrication of porous scaffolds for bone tissue engineering via low-temperature deposition. Scr Mater 46:771–776. CrossRefGoogle Scholar
  18. 18.
    Liu L, Xiong Z, Yan Y et al (2009) Multinozzle low-temperature deposition system for construction of gradient tissue engineering scaffolds. J Biomed Mater Res Part B Appl Biomater 88B:254–263. CrossRefGoogle Scholar
  19. 19.
    Bang Pham C, Fai Leong K, Chiun Lim T, Sin Chian K (2008) Rapid freeze prototyping technique in bio-plotters for tissue scaffold fabrication. Rapid Prototyp J 14:246–253. CrossRefGoogle Scholar
  20. 20.
    Yen HJ, Hsu SH, Tseng CS et al (2009) Fabrication of precision scaffolds using liquid–frozen deposition manufacturing for cartilage tissue engineering. Tissue Eng Part A 15:965–975. CrossRefGoogle Scholar
  21. 21.
    Kim G, Ahn S, Yoon H et al (2009) A cryogenic direct-plotting system for fabrication of 3D collagen scaffolds for tissue engineering. J Mater Chem 19:8817. CrossRefGoogle Scholar
  22. 22.
    Wang C, Zhou Y, Wang M (2017) In situ delivery of rhBMP-2 in surface porous shape memory scaffolds developed through cryogenic 3D plotting. Mater Lett 189:140–143. CrossRefGoogle Scholar
  23. 23.
    Tan Z, Parisi C, Di Silvio L et al (2017) Cryogenic 3D printing of super soft hydrogels. Sci Rep 7(1):16293. CrossRefGoogle Scholar
  24. 24.
    Liao C-Y, Wu W-J, Hsieh C-T et al (2016) Design and development of a novel frozen-form additive manufacturing system for tissue engineering applications. 3D Print Addit Manuf 3:216–225. CrossRefGoogle Scholar
  25. 25.
    Adamkiewicz M, Rubinsky B (2015) Cryogenic 3D printing for tissue engineering. Cryobiology 71:518–521. CrossRefGoogle Scholar
  26. 26.
    Mazur P (1984) Freezing of living cells—mechanisms and implications. Am J Physiol 247:C125–C142CrossRefGoogle Scholar
  27. 27.
    Mazur P (1970) Cryobiology: the freezing of biological systems. Science 168:939–949. CrossRefGoogle Scholar
  28. 28.
    D’Angelo G, Hansen HN, Hart AJ (2016) Molecular gastronomy meets 3D printing: layered construction via reverse spherification. 3D Print Addit Manuf 3:152–159. CrossRefGoogle Scholar
  29. 29.
    Dubey A (2016) How 3-D printing will shape food product development. Food Technol 70:53–61Google Scholar
  30. 30.
    Fernandez AID, Alvarez MAG (2017) Prevalence of dysphagia after stroke. View from primary care. Rqr Enferm Comunitaria 5:38–56Google Scholar
  31. 31.
    Cartwright A (2013) Time to recognise dysphagia as a contributing factor to malnutrition. Br J Community Nurs Suppl Nutr S6. CrossRefGoogle Scholar
  32. 32.
    Tobin A (2017) 3-D printed food for dysphagia sufferers. 3D Food Print Conf Asia-Pacific.
  33. 33.
    Leygonie C, Britz TJ, Hoffman LC (2012) Impact of freezing and thawing on the quality of meat: review. Meat Sci 91:93–98CrossRefGoogle Scholar
  34. 34.
    Blossey R (2003) Self-cleaning surfaces—virtual realities. Nat Mater 2:301–306. CrossRefGoogle Scholar
  35. 35.
    Ren S, Yang S, Zhao Y et al (2003) Preparation and characterization of an ultrahydrophobic surface based on a stearic acid self-assembled monolayer over polyethyleneimine thin films. Surf Sci 546:64–74. CrossRefGoogle Scholar
  36. 36.
    Gergely RCR, Pety SJ, Krull BP et al (2015) Multidimensional vascularized polymers using degradable sacrificial templates. Adv Funct Mater 25:1043–1052. CrossRefGoogle Scholar
  37. 37.
    Sura L, Madhavan A, Carnaby G, Crary MA (2012) Dysphagia in the elderly: management and nutritional considerations. Clin Interv Aging 7:287–298Google Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  • Bartłomiej Zawada
    • 1
  • Gideon Ukpai
    • 1
    Email author
  • Matthew J. Powell-Palm
    • 1
  • Boris Rubinsky
    • 1
  1. 1.Department of Mechanical EngineeringUniversity of California BerkeleyBerkeleyUSA

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