Design Innovation

Development of a Three-Dimensional Bioprinter: Construction of Cell Supporting Structures Using Hydrogel and State-Of-The-Art Inkjet Technology

[+] Author and Article Information
Yuichi Nishiyama, Chizuka Henmi, Kumiko Yamaguchi, Shuichi Mochizuki, Hidemoto Nakagawa, Koki Takiura

 Kanagawa Academy of Science and Technology, Think-E Building, 1-23, Minamiwatarida, Kawasaki-ku, Kawasaki 210-0855, Japan

Makoto Nakamura1

Graduate School of Science and Engineering for Research (Engineering), University of Toyama, 3190, Gofuku, Toyama 930-8555, Japan; Kanagawa Academy of Science and Technology, Kanagawa, Japanmaknaka@eng.u-toyama.ac.jp


Corresponding author.

J Biomech Eng 131(3), 035001 (Dec 31, 2008) (6 pages) doi:10.1115/1.3002759 History: Received March 06, 2007; Revised May 16, 2008; Published December 31, 2008

We have developed a new technology for producing three-dimensional (3D) biological structures composed of living cells and hydrogel in vitro, via the direct and accurate printing of cells with an inkjet printing system. Various hydrogel structures were constructed with our custom-made inkjet printer, which we termed 3D bioprinter. In the present study, we used an alginate hydrogel that was obtained through the reaction of a sodium alginate solution with a calcium chloride solution. For the construction of the gel structure, sodium alginate solution was ejected from the inkjet nozzle (SEA-Jet™, Seiko Epson Corp., Suwa, Japan) and was mixed with a substrate composed of a calcium chloride solution. In our 3D bioprinter, the nozzle head can be moved in three dimensions. Owing to the development of the 3D bioprinter, an innovative fabrication method that enables the gentle and precise fixation of 3D gel structures was established using living cells as a material. To date, several 3D structures that include living cells have been fabricated, including lines, planes, laminated structures, and tubes, and now, experiments to construct various hydrogel structures are being carried out in our laboratory.

Copyright © 2009 by American Society of Mechanical Engineers
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Figure 1

Alginate gel beads

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Figure 2

Gel beads including HeLa cells

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Figure 3

Comparison of inkjet droplets and bovine endothelial cells

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Figure 4

EPSON SEA-Jet™ inkjet nozzle head

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Figure 5

Newly developed bioprinter: (a) 3D bioprinter developed in our laboratory and (b) 3D bioprinter in a bioclean bench environment

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Figure 6

Illustration of the gel structure fabrication method

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Figure 7

Gel lines formed using the 3D bioprinter (nozzle head speed: 35mm∕s, ejecting frequency: 500Hz): (a) alginate gel solid line (nozzle head speed: 40mm∕s, ejecting frequency: 1kHz) and (b) alginate gel broken line

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Figure 8

Gel sheet (nozzle head speed: 25mm∕s, ejecting frequency: 800Hz)

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Figure 9

Gel lamination (nozzle head speed: 25mm∕s, ejecting frequency: 800Hz)

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Figure 10

Gel tube with enlarged view of the opening at the tip of the tube (nozzle head speed: 25mm∕s, ejecting frequency: 800Hz)

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Figure 11

Gel tube including HeLa cells (nozzle head speed: 25mm∕s, ejecting frequency: 800Hz)

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Figure 12

Viscosity of the sodium alginate and PVA solutions determined using a viscometer: (a) sodium alginate solution and (b) PVA solution

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Figure 13

Viscosity of the fibrinogen and hyaluronan solutions determined using a viscometer: (a) fibrinogen solution and (b) hyaluronan solution




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