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RESEARCH ARTICLE

A novel 3D printing method for cell alignment and

differentiation

1*
1
1†
1†
Ramya Bhuthalingam , Pei Qi Lim , Scott A Irvine , Animesh Agrawal , Priyadarshini S
1
2
1
2
Mhaisalkar , Jia An , Chee Kai Chua and Subbu Venkatraman
1 Materials and Science Engineering, Nanyang Technological University, N4.1-01-30, 50 Nanyang Avenue, Singapore
639798, Singapore
2 Singapore Centre for 3D printing, School of Mechanical & Aerospace Engineering, Nanyang Technological University,
50 Nanyang Avenue, Singapore 639798, Singapore

Abstract: The application of bioprinting allows precision deposition of biological materials for bioengineering applica-
tions. Here we propose a 2 stage methodology for bioprinting using a back pressure-driven, automated robotic dispens-
ing system. This apparatus can prepare topographic guidance features for cell orientation and then bioprint cells directly
onto them. Topographic guidance features generate cues that influence adhered cell morphology and phenotype. The
robotic dispensing system was modified to include a sharpened stylus that etched on a polystyrene surface. The same
computer-aided design (CAD) software was used for both precision control of etching and bioink deposition. Various
etched groove patterns such as linear, concentric circles, and sinusoidal wave patterns were possible. Fibroblasts and
mesenchymal stem cells (MSC) were able to sense the grooves, as shown by their elongation and orientation in the di-
rection of the features. The orientated MSCs displayed indications of lineage commitment as detected by fluores-
cence-activated cell sorting (FACS) analysis. A 2% gelatin bioink was then used to dispense cells onto the etched fea-
tures using identical, programmed co-ordinates. The bioink allows the cells to contact sense the pattern while containing
their deposition within the printed pattern.
Keywords: bioprinting, surface guidance, automated robotic deposition, precision etching, bioink, stem cells

*Correspondence to: Scott A Irvine, Materials and Science Engineering, Nanyang Technological University, N4.1-01-30, 50 Nanyang
Avenue, Singapore 639798, Singapore; Email: sairvine@ntu.edu.sg
† The first two authors contributed equally to this paper.

Received: May 19, 2015; Accepted: June 24, 2015; Published Online: July 2, 2015
Citation: Bhuthalingam R, Lim P Q, Irvine S A, et al. 2015, A novel 3D printing method for cell alignment and differentiation. In-
ternational Journal of Bioprinting, vol.1(1): 57–65. http://dx.doi.org/10.18063/IJB.2015.01.008.

[3]
1. Introduction of the attached cells . Cues can be introduced to the
R coordinates the attachment of the cell anchoring inte-
surface of a cell binding material that attracts and
obotic deposition has a promising potential
in biomaterial engineering for automated and
[4,5]
grin proteins
. The shape adopted by the cell thr-
reproducible patterning for cell delivery. Its
application has given tissue engineers a method for ough the interaction with the surface cues feeds back
to the cell nucleus via integrin receptors, which in turn
cellular deposition with control up to the micrometer affects the gene expression profile of the adhered
level [1,2] . Additionally, cell adhering surfaces can be cell [6,7] . This can have the effect of influencing cellu-
modified to influence the orientation and morphology lar behavior, such as stimulating the switch in smooth

A novel 3D printing method for cell alignment and differentiation © 2015 Ramya Bhuthalingam, et al. This is an Open Access article distributed
under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/),
permitting all non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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