Rock Star Student

Ashley Rea
ENC 2135
Date:________

3D bioprinting is a technique where stem cells are used to create a 3-dimentional,

functional organ. This process is complicated and relies on the use of stem cells, a highly
controversial topic in society. The primary focus of this paper will be to discuss the 3D
bioprinting process. To achieve the goal of discussing 3D bioprinting, this paper will talk about
stem cells and other materials used in the 3D bioprinting process, as well as the process itself.

I. 3D Bioprinting: A Brief Overview:

3-D bioprinting is a computer-aided manufacturing process based off of traditional 3D printing
that deposits living cells together with hydrogel-based scaffolds and allows for patterning of
individual components of the tissue or organ thereby facilitating formation of complex tissue
architecture (Varkey, Atala 3). By using stem cells as an ink of sorts in a 3D printer like device,
differentiated cells can be printed in the form of an organ by replicating a 3D model from a
computer. Bioprinting is described in the paper published by Murphy and Atala as a process
where layer-by-layer precise positioning of biological materials, biochemical and living cells,
with special control of the placement of functional components, is used to fabricate 3D
structures (Murphy, Atala 1).

II. Stem Cells:

Before the 3D bioprinting process can be discussed, it is necessary to take a look at the
two primary types of stem cells, which are essential to the creation of functional tissue made
through the bioprinting process. To begin, stem cells are unique cell populations with the ability
to undergo both self-renewal and differentiation into different lineages, which means that they
are cells that can divide indefinitely and differentiate into any specific cell type they want
(Biswas, Hutchins 1). Stem cells can differentiate into any cell type, meaning the can be
programmed to become muscular tissue, skeletal, skin, and even cardiac tissue among other
things.
The first, and most commonly known type of stem cell due to the controversy that
surrounds it, is embryonic stem cells, or ES cells. ES cells are cells [that] have the ability to
form any fully differentiated cell of the body (Biswas, Hutchins 1). The ability to become any
type of cell type is what makes embryonic stem cells versatile, more so than other types of stem
cells, and is the reason why theyre so sought after. ES cells are also desirable because of the fact
that the tissue created from them can be transplanted to any patient without worry of rejection
from the body. ES cells can only be cultivated in a narrow window of 5-9 days after conception
from the embryo (Biswas, Hutchins 2). There are three common methods for differentiating
undifferentiated embryonic stem (ES) cells: (1) co-culturing ES cells with inducing cells, (2)
treating cultures with growth factors, or (3) a combination of both (Biswas, Hutchins 4).
The second, lesser known, but more ethical type of stem cell is an induced pluripotent
stem cell, or iPS cell, which comes from reprogrammed adult cells that come from the patient.

iPS cells are considered more ethical than ES cells due to the fact that the cells come from the
patient, meaning that theres no destruction of an embryo as there is with ES cells. iPS cells are
different from ES cells in the fact that they are essentially differentiated cells that are
reprogrammed to an embryonic-like state by transfer of nuclear contents into oocytes or by
fusion with embryonic stem (ES) cells (Takahashi, Yamanaka 1). Since iPS cells are created
from the cells of the patient, meaning they are a genetic match to the patient, there is no risk of
the tissue transplantation being rejected by the bodys immune system, which solves one of the
biggest problems with organ transplants today. iPS cells seem like a better type of cell to use for
3D bioprinting, since they do the same thing as ES cells without the ethical concerns, however
there is one major issue with iPS cells that we havent solved yet: cancer. iPS cells can become
cancerous, and more research needs to be conducted before the risk of cancer in iPS cells can be
addressed properly (Daley 1).
The process for creating an iPS cell takes much longer and difficult than that of ES cells.
Instead of extracting cells from an embryo, iPS cells have the nuclear contents, DNA and
nucleoplasm, are put into an oocyte, a type of cell found in an ovum, also known as an
unfertilized female egg (Takahashi, Yamanaka 1). In other words, the nucleus of a cell contains
all the programming so to speak to make cells that are exact copies of itself. The nucleus is then
put into an oocyte where it is allowed to divide indefinitely using instructions from the nucleus
implanted in the oocyte, which creates the same type of tissue that the nucleus came from. Add
an example of what Im talking about here?

III. Materials and Scaffolds:

Initially, 3D printing technologies were designed for nonbiological applications, such as
the deposition of metals, ceramics and thermoplastic polymers, and involved the use of organic
solvents and high temperatures which are not compatible with biological cells (Murphy, Atala 6).
In the paper 3D bioprinting of tissue and organs, Murphy and Atala discuss one of the major
challenges of 3D bioprinting: to find materials that are not only compatible with biological
materials and the printing process but can also provide the desired mechanical and functional
properties for tissue constructs (Murphy, Atala 6). Finding materials that can be used in 3D
bioprinting are so difficult because they must meet strict criteria as pointed out by Ozblat, I. T.
and Yin Yu in a paper on bioprinting toward organ fabrication Both synthetic and natural
polymers have been used to engineer various tissue grafts like skin, cartilage, bone, and bladder.
To be successfully used for tissue engineering, these materials must be biocompatible and
biodegradable, with the mechanical strength to support cell attachment, proliferation, and direct
cell differentiation toward certain lineages (Ozblat, Yu 1).
SCAFFOLDS (HYDROGELS)

IV. The 3D Bioprinting Process:

3D bioprinting is based on three central approaches: biomimicry, autonomous selfassembly, and mini-tissue building blocks (Murphy, Atala 1). The first approach, biomimicry, is
where technology is inspired by biological factors and characteristics. (1) Biomimicry is
achieved in 3D bioprinting by manufacturing an identical reproduction of the cellular and
extracellular components of a tissue or organ (Murphy, Atala 1). For this approach to succeed,

the replication of biological tissues on the microscale is necessary (Murphy, Atala 2). (2) The
second approach to bioprinting is autonomous self-assembly. In this approach, embryonic organ
development is used as a guide for replicating tissue. This would mean allowing cells to create
their own extracellular matrix components, which allows cells to self-assemble and manipulation
of the environment to get cells to grow in the desired manner. (3)The final approach, minitissues, involves defining the smallest structural and functional component of a tissue. After the
component is defined, that component is printed repeatedly, and allowed to self-assemble into
organs.
Before an organ or tissue can be printed using one of the approaches discussed in the
previous paragraph, it is necessary to create a high definition 3D model for which the printing of
the organ will be based off of. This is primarily done by using noninvasive imaging technologies,
such as an MRI or CT scan, and importing that information into a Computer-aided design and
computer aided manufacturing (CAD-CAM) technology coupled with mathematical modeling to
create the 3D model that will be used as the blueprint for the tissue being printed. The process of
imaging and modeling a tissue/organ is known as transformation of analytical anatomy to
synthetic anatomy (Murphy, Atala 3). Something interesting to note is that a 3D model of a
patients own organ may not be desirable due to disease or injury, and the fact that its not
economically feasible for large-scale production of organs.
In a paper by Varkey and Atala discussing organ bioprinting and ethics/policies, they
describe the 3D bioprinting processes a simple manner that encompasses what the process is like.
Varkey and Atala describe the 3D bioprinting process for fabricating tissue as a layer-by-layer
addition of material on a preshaped supporting scaffold to build a 3-D tissue or organ construct
with input from a computer-aided design file (Varkey, Atala 4). The CAD file determines exactly

where everything goes and even places the growth factors necessary for tissue regeneration.
----------Expand upon this----

V. Tissue Bioprinting Strategies:

There are the main technologies used for the deposit and pattering of biological materials:
inkjet, microextrusion, and laser-assisted printing. Each type of printing technology has its own
range of capabilities depending on what needs to be printed. Different features of these
technologies should be considered in light of the most important factors in 3D bioprinting:
surface resolution, cell viability, and the biological materials used for printing (Murphy, Atala 3).
Inkjet bioprinting, also known as drop-on-demand printers, are the most common type
of bioprinter. It works by having controlled volumes of liquid being delivered to predefined
locations. Inkjet printers use thermal or acoustic forces to eject the bio-ink onto a substrate,
which can support or form part of the final construct.
Thermal inkjet printers function by heating the print head to produce pressure that forces
the bio-ink out of the nozzle, which can make the tip reach temperatures as high as 300 C (572
F), which has surprisingly been found to have no significant impact on the structure of the tissue
being printed nor on the DNA within the cells being extruded. There are disadvantages to using
thermal inkjet printers, since they incur the risk of exposing cells and materials to thermal
and mechanical stress, low droplet directionality, nonuniform droplet size, frequent
clogging of the nozzle and unreliable cell encapsulation (Murphy, Atala 4).

Acoustic inkjet bioprinters contain a piezoelectric crystal that vibrates due to a rapid
change in size when electricity is run through it. The vibration from the nozzle ejects the bio-ink

as droplets at regular intervals. The amount of electricity run through the crystal can be used to
control the parameters, such as pulse, duration and amplitude, can be adjusted to
control the size of droplets and the rate of ejection. Advantages of acoustic inkjet
printers include the capability to generate and control a uniform droplet size and
ejection directionality as well as to avoid exposure of cells to heat and pressure
stressors (Murphy, Atala 4). Finish up acoustic inkjet, --discuss what inkjet
printers are most suitable to print--

--Discuss microextrusion
--Discuss laser assisted printing
--impact on society (perhaps include in introduction?)
--conclusion--

Ethics?
Example of 3D printing via paper where muscle and bone was printed?

Works Cited
Biswas, Atindriya, and Robert Hutchins. "Embryonic Stem Cells." Stem cells and
development 16.2 (2007): 213-21. Web.

Kang, Hyun-Wook, et al. "A 3D Bioprinting System to Produce Human-Scale Tissue Constructs
with Structural Integrity." Nat Biotech advance online publication (2016)Print.
Kolesky, David B., et al. "3D Bioprinting of Vascularized, Heterogeneous Cell-Laden Tissue
Constructs." Advanced Materials 26.19 (2014): 3124-30. Print.
Murphy, Sean V., and Anthony Atala. "3D Bioprinting of Tissues and Organs." Nature
biotechnology 32.8 (2014): 773-85. Print.
Ozbolat, Ibrahim. "Special Issue: 3D Bioprinting." Journal of nanotechnology in engineering
and medicine(08)Print.
Ozbolat, I. T., and Yin Yu. "Bioprinting Toward Organ Fabrication: Challenges and Future
Trends." Biomedical Engineering, IEEE Transactions on 60.3 (2013): 691-9. Print.
Pereira, Rben F., and Paulo J. Brtolo. "3D Bioprinting of Photocrosslinkable Hydrogel
Constructs." Journal of Applied Polymer Science 132.48 (2015): n/a,n/a. Print.
Singh, Deepti, Dolly Singh, and Soo Han Sung. "3D Printing of Scaffold for Cells Delivery:
Advances in Skin Tissue Engineering." Polymers (20734360) 8.1 (2016): 1-17. Print.
Takahashi, Kazutoshi, and Shinya Yamanaka. "Induction of Pluripotent Stem Cells from Mouse
Embryonic and Adult Fibroblast Cultures by Defined Factors." Cell 126.4 (2006): 663-76.
Print.
Varkey, Mathew, and Anthony Atala. "Organ Bioprinting: A Closer Look at Ethics and
Policies." Wake Forest Journal of Law & Policy 5 (2015): 275-98. Print.