Available online at www.sciencedirect.
com Current Opinion in
ScienceDirect
3D bioprinting from the micrometer to millimeter
length scales: Size does matter
T. J. Hintona,1, Andrew Leea,1 and Adam W. Feinberga,b
3D bioprinting has rapidly expanded from a niche technology
and in to a versatile platform for fabricating tissues with com-
plex geometries and features ranging from the cellular to organ
length scales. Recent advances include the development of
yield-stress support baths that enable soft hydrogels to be 3D
printed in highly complex structures and vascular networks and
successful implantation of 3D printed scaffolds in animal
models and human patients towards regenerating bone,
cartilage and muscle. However, the range of additive
manufacturing technologies currently used each has distinct
advantages and disadvantages, and specifically it is critical to
understand how the resolution of these different approaches
dictate structure and function of the engineered tissue con-
structs. This article offers a focused review on how the size of
the biomaterial that is being deposited or polymerized, which
can range over 4 orders-of-magnitude from ~1 mm up to 1 mm,
drives biological performance.
Addresses
a
Department of Biomedical Engineering, Carnegie Mellon University,
Pittsburgh, PA, USA
b
Department of Materials Science and Engineering, Carnegie Mellon
University, Pittsburgh, PA, USA
Corresponding author: Feinberg, Adam W (feinberg@andrew.cmu.edu)
1
Denotes equal contribution.
Current Opinion in Biomedical Engineering 2017, 1:31–37
This review comes from a themed issue on Future of BME
Edited by George Truskey
Received 16 December 2016, revised 7 February 2017, accepted 13
February 2017
http://dx.doi.org/10.1016/j.cobme.2017.02.004
2468-4511/© 2017 Published by Elsevier Inc.
Abbreviations
FDM, fused deposition modeling; SLS, selective laser sintering; AM,
additive manufacturing; MRI, magnetic resonance imaging; CT,
computed tomography; FRESH, Freeform Reversible Embedding of
Suspended Hydrogels; DLP, digital light processing; PCL, poly-
caprolactone; PEKK, polyetherketoneketone.:
Introduction
Over the past decade, 3D bioprinting has rapidly
evolved from a niche research area and into a main-
stream biofabrication platform. Core to this shift is the
robotic-controlled, additive manufacturing process that
www.sciencedirect.com
Biomedical Engineering
enables precision layer-by-layer material deposition in 3
dimensions (Figure 1a). The cost of these systems has
decreased significantly, driven by open-source hardware
and software technologies that have enabled develop-
ment of new bioprinting approaches and spurred broader
adoption through custom-built and lower-cost com-
mercial printers [1e3]. Equally important is the ability
to use patient-specific 3D medical imaging data from
MRI and CT, providing the potential to repair regions of
damaged or diseased tissues, and perhaps eventually
whole organs [4].
Herein, we describe recent advances in 3D bioprinting,
with a focus on how the physical size (resolution) of the
material being deposited dictates the performance
and properties of the scaffolds being fabricated,
and ultimately the functionality of the engineered
tissues. This is important because 3D bioprinting en-
compasses multiple underlying technologies that achieve
additive manufacturing (AM) through extrusion, photo-
polymerization, melting and/or jetting. Resolution can
range from w1 mm diameter for extruded filaments down
to <1 mm diameter photopolymerized voxels, with each
approach and material used representing a trade off in
resolution, print time, and cell viability (Figure 1b, c).
Researchers have now demonstrated this across a broad
range of applications from large-volume constructs for
tissue repair to microvascular networks for improved cell
viability. It is clear that size does matter, with distinct
advantages and disadvantages for scaffolds fabricated at
the micrometer, mesoscopic and millimeter length scales.
How these factors interrelate is critical to understanding
current and future advances in 3D bioprinting.
3D bioprinting with millimeter-scale
features for large-volume scaffolds
3D bioprinting has emerged as a viable strategy for
fabricating large-volume constructs with anatomically-
accurate structure. The goal for constructs printed at
this scale are to one day be transplantable into humans
to replace diseased tissues and organs. To this end, the
additive manufacturing approaches that have achieved
the most clinically-relevant results in 3D bioprinting
are extrusion-based deposition and laser-based tech-
nologies that have already had decades of use with
industrial-grade plastics. For example, both extrusion-
based deposition of thermoplastics, commonly known
as fused deposition modeling (FDM), and laser-based
fusion of thermoplastic powders, commonly known as
selective laser sintering (SLS), are mature 3D printing
Current Opinion in Biomedical Engineering 2017, 1:31–37
32 Future of BME
Figure 1
(a) Schematic of a 3D bioprinter consisting of a 3-axis robot with one or more fluid extruders. (b) Extrusion nozzles vary in diameter orders of magnitude
from approximately 10 mm–1 mm, and focused laser light to a sub-micron spot size smaller than a cell. (c) For comparison, living tissue such as skeletal
muscle possesses hierarchical organization that spans several orders of magnitude from the macro-to microscopic scale.
technologies that have transitioned into 3D bioprinting
by using biocompatible polymers. These approaches
have an effective resolution of approximately 400 mm to
1 mm depending on the orifice diameter of the ther-
moplastic extruder in FDM or the powder diameter and
laser spot size for SLS. While this results in limited
control of material microstructure, it provides distinct
advantages in fabricating large constructs >1 cm3 in
volume, and as a result has been the primary approach
used for in vivo studies. For example, SLS 3D printing of
polyetherketoneketone (PEKK) is used for its high
strength in the load-bearing function of a 3D printed
cranial plate for human skull reconstruction surgeries
[5]. This demonstrates that 3D bioprinting of non-
resorbable, medical-grade polymers can produce FDA-
approved, high-performance medical devices that can
integrate with patient-specific anatomy. The future of
3D bioprinting lies in extending this to biodegradable
materials that can support true tissue regeneration. For
example, Zopf et al. SLS printed a tracheal splint from
polycaprolactone (PCL) to support ventilation in a pe-
diatric patient with respiratory failure, utilizing the
relatively high stiffness material to prevent tracheal
collapse and a porous design for flexibility during
respiration (Figure 2aec) [6]. The PCL is a biode-
gradable thermoplastic, and depending on formulation,
can be tuned to degrade over a period of months to years
to match the rate of tissue regeneration. These exam-
ples serve to demonstrate the feasibility of adapting
traditional 3D printing technologies for creating rigid
constructs with structurally functional millimeter-scale
features.
For 3D bioprinting soft tissue, recent advances have
focused on incorporating cells and soft biomaterials such
as hydrogels and evaluating performance in more complex
in vitro and in vivo environments. This has largely involved
Current Opinion in Biomedical Engineering 2017, 1:31–37
depositing a range of hydrogels using syringe-based ex-
truders that function similar to FDM, but avoid the
thermal heating that can damage cells and biological
molecules. For example, Manoor et al. 3D printed an ear
using a cell-laden alginate hydrogel bioink that could
receive auditory signals via its embedded conductive coils
(Figure 2d) [7], essentially creating a biohybrid device.
Using a similar approach, Rhee et al. 3D printed a cell-
laden collagen ink in the shape of the meniscus cartilage
and found that the construct was mechanically stable with
tunable compressive strength (Figure 2e) [8]. However,
the resolution of hydrogels 3D printed in air is limited by
the deformability of these materials. In order to create
constructs that could better maintain geometry and be
implanted in vivo, multi-extruder setups were used by
both Kang et al. and Daly et al. to sequentially deposit a
PCL thermoplastic alongside a stem-cell laden hydrogel
ink to create bone-like constructs [9,10]. When implanted
subcutaneously, the PCL served as a structural support,
and the hydrogel components were found to be osteoin-
ductive throughout the millimeter-scale constructs.
Hung et al. showed that PCL can be further modified with
decellularized bone matrix particles to facilitate improved
bone regeneration in a temporomandibular joint construct
(Figure 2f, g) [11]. Tissue-specific maturation strategies
can also be incorporated into printed constructs based on
print geometry. Kang et al. also combined cell-laden
hydrogel ink, PCL and Pluronic F-127 as a sacrificial ma-
terial to temporarily support parallel extrusions of a
hydrogel-myoblast mixture, facilitating myoblast align-
ment and fusion in culture (Figure 2h, i) [9]. When
implanted for 2 weeks in a mouse model, the construct
showed a degree of neural and vascular integration. These
studies add to a growing body of research that shows the
feasibility of macroscale bioprinting for fabricating large
anatomically-accurate constructs, and the potential for
in vivo applications.
www.sciencedirect.com
 3D bioprinting from the micrometer to millimeter Hinton et al. 33

Figure 2

(a) Minimum intensity projection of the respiratory airway from a CT scan is used to (b) 3D print a tracheal splint (red box) and its intended placement
location on a similarly printed model of the airway. (c) The splint is placed and sutured over the softened region of the left mainstem bronchial segment
[6]. (d) Bioprinting of a bionic ear with a chondrocyte-seeded alginate hydrogel and silver nanoparticle infused conductive coil [7]. (e) 3D printed
meniscus from collagen I hydrogel [8]. (f) The temporomandibular joint of the jaw (circled) is (g) bioprinted with varying percentage of decellularized
mineral particles [11]. (h) Muscle fiber bundle design is (i) printed with a sacrificial material Pluronic F-127 for temporary support of a myoblast-laden
hydrogel [9].

3D bioprinting at the millimeter scale has achieved
important advances in fabricating organ-like constructs
that can be implanted into animal models and human
patients. However, higher resolution features are
necessary for recapitulating structure and ultimately
function of native tissues. While these features can be
achieved with the use of a smaller extruder orifice or
finer laser focus, smaller extrusion diameters lead
directly to a larger number of machine movements,
which results in longer print times. For example, an
organ-scale construct can take 2e10 h to print, which is
problematic because cell viability will decrease and cells
can settle out of suspension and alter uniformity of the
bioink [12,13]. Although increasing print speeds can
reduce print times of these large-scale constructs, high
speed and high precision movement systems are chal-
lenging to implement. Biomaterials must also be
developed specifically for printability, which is dictated
by material rheology and gelation mechanism, while also
maintaining tissue-specific biochemical and mechanical
properties in the printed construct. While Pati et al.
have shown that it is possible to create organ-derived
extracellular matrix (ECM) inks from animal tissues,
immunogenicity of these inks as well as material abun-
dance are major hurdles when scaling towards human-
sized constructs [14]. Until we can interface large
organotypic structures with higher resolution features at
the meso- and microscale without loss of cell viability,
macroscopic printed constructs will be able to reproduce
the tissue geometry but be limited in terms of tissue-
specific function.
3D bioprinting at the mesoscale to control
hierarchical architecture
Within tissue engineering, the mesoscale bridges the
macro- and microscopic domains and thus provides the
ability to define specific hierarchical architectures [15].
Branching vascular networks are key mesoscale features
important for providing nutrient supply in thick, tissue
engineered constructs, but they have traditionally been
difficult to construct. Recent technological advances
have uniquely positioned 3D bioprinting as a feasible
strategy for fabricating these structures at physiologically-
relevant resolutions.
3D printed sacrificial (i.e., fugitive) materials that can
be removed later have proven to be highly effective for
predefining void regions within a bulk gel for micro-
vascular networks. For example, multiple groups have
printed sacrificial materials such as thermosensitive
Pluronic F-127 ink or water-soluble carbohydrate glass,
casted an ECM gel around the printed structure, and
removed the sacrificial material to produce mesoscale
vascular-like structures [16e19]. This approach allows
fabrication of intricate 3D void networks that resemble
key physiological structures that are otherwise difficult
to manufacture, such as a thick vascularized tissue
(Figure 3a, b), a convoluted renal proximal tubule model
(Figure 3c, d) [17,18] and a vascular lattice network
(Figure 3eeg) [19]. Instead of patterning the void re-
gions of a construct with a sacrificial material, Itoh et al.
defined the 3D position of cell spheroids w400 mm in
diameter by immobilizing them on an array of metal

www.sciencedirect.com Current Opinion in Biomedical Engineering 2017, 1:31–37

34 Future of BME

Figure 3

(a) A vascularized tissue fabrication process that casts an ECM hydrogel around printed paths of a sacrificial ink, which can be evacuated and perfused
to create (b) a thick multicellular tissue with endothelialized vascular network [17]. A similar technique is used to produce a (c) renal proximal tubule
model with (d) a dense cellular monolayer with native-like morphological features such as the cilia and functional epithelial marker Na+/K+ ATPase [18].
(e) Schematic and (f) perfusion time-lapse of vascular channels fabricated from fugitive carbohydrate glass [19]. (g) An array of metal needles used to
spatially position pre-formed cell spheroids in 3D to create (h) a compacted, cellularized tubular structure [20]. (i) Freeform embedded printing into
thermoreversible support to form (j) single alginate hydrogel strand, (k) a perfusable vascular tree network, and (l) a 3D printed heart from an embryonic
chick heart model [23]. (m) Embedded printing of a (m) fluorescent microsphere suspension and (n) polyvinyl alcohol into a granular gel medium to
create complex 3D structures [24].

needles (Figure 3g, h) [20]. Over time the cell spheroids
fuse into an integrated tissue, with resolution limited by
spheroid diameter and the void space generated by
spheroid separation. These approaches demonstrate the
importance of patterning void space in 3D, but do not
address the mesoscale printing of hydrogels directly.
Inkjet is another 3D bioprinting technology that can
deposit hydrogels and cell directly in a drop-wise
fashion. Recent work has demonstrated the 3D print-
ing of silk fibroin using a sacrificial alginate support [21]
and collagen combined with thermo-responsive agarose
for bone tissue engineering applications [22]. While an
important approach, inkjet mesoscale printing requires a
support material, or special ink formulations that
incorporate a sacrificial component, to deposit soft
hydrogels at high resolution.
3D bioprinting of soft hydrogels within a support bath can
achieve mesoscale material deposition to build complex
scaffolds that would deform if unsupported. Using a
method termed Freeform Reversible Embedding of
Suspended Hydrogels (FRESH), Hinton et al. showed
that a hydrogel precursor solution could be deposited and
gelled within a thermoreversible support bath composed
of a gelatin microparticulate slurry (Figure 3i, j) [23].
Complex 3D prints of alginate, fibrin, Matrigel, and
collagen were achieved, including a perfusable model of a
human right coronary artery tree from MRI (Figure 3k)

Current Opinion in Biomedical Engineering 2017, 1:31–37 www.sciencedirect.com

 3D bioprinting from the micrometer to millimeter Hinton et al. 35

and an embryonic-stage, trabeculated chick heart from biological function and complexity. As 3D printers evolve,
multiphoton imaging (Figure 3l) [23]. Unique to the these systems will begin operating with advanced motion
FRESH process is complete melting of the support bath control, microliter extrusion precision, and increased
at a physiologic 37  C, and bioprinted sheets of cells in resolution, which combined with metrological validation,
collagen-Matrigel hydrogels showed viability greater than will improve the output of mesoscale bioprinting.
97% [23]. Bhattacharjee et al. reported a similar approach
using a slurry (granular medium) of carbomer microgels as
3D bioprinting at the microscale to engineer
a support bath for a variety of UV-curable inks with
the cellular environment
embedded cells [24] and even the direct 3D printing of
It is now recognized that the structure of the ECM on the
cells alone [25]. Complex 3D prints were fabricated from
subcellular level influences cell behavior and tissue or-
fluorescent inks embedded into the support bath,
ganization. However, using extrusion to 3D bioprint
including a 3D tube with a complicated knot geometry
tissue microarchitecture is challenging because it re-
(Figure 3m) and a jellyfish inspired print with intricate
quires handling small volumes of non-Newtonian
arms (Figure 3n) [24]. These embedded printing ap-
biopolymer fluids. For this reason, digital light process-
proaches have expanded the complexity of 3D bioprinted
ing (DLP) and multiphoton laser scanning have emerged
scaffolds from hydrogels, but there are challenges to
as 3D bioprinting platforms for photocrosslinking bio-
depositing one material within a second material. For
materials and building scaffolds with spatial resolutions of
example, the granular support bath can influence the
<10 mm. For example, Ma et al. have demonstrated that
morphology of the deposited material and the thermo-
DLP printing and visible-light photo-initiators can be
reversible microparticles can also leave void space
used to develop constructs of multiple cell types and
within the print once removed [3,23]. Future studies will
materials at a scale consistent with that of the human liver
need to evaluate how cells will respond to this surface
lobule (Figure 4a, b) [26]. Claeyssens et al.utilized
topography or how this porosity will affect long term cell
multiphoton excitation to print PCL-based scaffolds with
culture of printed scaffolds.
4 mm features, and Zhang et al. were able to pattern
PEGDA into 1 mm spheres and filaments [27,28]. Arrays
Maturing printed tissues in culture will require consid-
of filament-like polymerized resin created by Ma et al.
eration of nutrient transport and cell-driven compaction
were able to align contractile cardiomyocytes (Figure 4c,
as part of printed tissue design; moreover, printing pa-
d, e) [29]. The multiphoton effect can also be used to
rameters such as infill patterning or spacing between
selectively stiffen a gel (Figure 4f) or spatially pattern
perimeter extrusions may be critical for achieving desired
molecules at specific locations in 3D (Figure 4g) [30,31].
properties. Merging the multiple process parameters of
These optical printing methods not only create micro-
extrusion-based 3D printing with a better understanding
scopic hydrogel geometries, but also demonstrate the
of engineered tissue design has the potential for improved

Figure 4

(a) Digital light processing (DLP) engines with individually addressed pixels can render complex patterns of illumination onto a variety of substrates, and
(b) patterned photopolymerization can selectively isolate different cell types in separate gels [26]. (c) Multiphoton photopolymerization can be used to
scan a small excitation point through a medium creating (d) rods of polymer that can support (e) cultures of aligned cardiomyocytes [29]. (f) Hollow
structures fabricated via multiphoton photopolymerization [30]. (g) Fluorescent image of separate growth factor molecules (red and green) photo-
immobilized by multiphoton photopolymerization within a 3D hydrogel [31].

www.sciencedirect.com Current Opinion in Biomedical Engineering 2017, 1:31–37

36 Future of BME
ability to spatially pattern molecular concentration and
matrix stiffness within these gels.
Light-based techniques achieve high resolution but need
to address the settling of cells out of photopolymer sus-
pensions, phototoxicity from repeated irradiation, and the
time required to prepare each new layer for exposure to
light. For example, in DLP and similar projection-based
methods, liquid prepolymer must be added to build the
next layer, which takes a relatively long time compared to
the duration of exposure and can produce shearing forces
on cells. Similarly, as print time increases exponentially
with higher resolution, cell viability decreases due to time
in suspension and outside of standard culture conditions.
Continuous Light Interface Printing (CLIP) and other
light-based continuous exposure printing methods may
offer hope for faster, higher resolution bioprinting, but the
chemistry is not currently biocompatible [32]. Scaling
high-resolution microscopic techniques up toward the
macroscopic requires extremely precise hardware and the
entire process needs to be engineered to keep cells in
suspension alive during the extended print time. Looking
forward, faster and more precise hardware will be required
to scale microscopic bioprinting to a macroscopic additive
manufacturing process.
Conclusions
Currently, 3D bioprinting broadly attempts to utilize
mesoscale manufacturing technologies to achieve
microscopic control of cells and matrix across millimeter
length scales. While millimeter-scale 3D bioprinting
techniques are well-characterized and mature enough to
have animal and human implantation models, they cannot
control the microstructures responsible for directing
biological events like cell alignment or matrix remodeling.
Mesoscale bioprinting methods represent the current
thrust of research efforts, allowing many to obtain con-
structs with perfusable vasculatures and complex,
branching architectures. Techniques with micrometer-
scale accuracy and precision are currently light-based,
making them limited in scope to materials that are
photopolymerizable. Unfortunately, there is no tech-
nique that spans from the microscopic to macroscopic
scales and spans materials while 3D printing with cells in
complex geometries.
While we cannot address all of these challenges at once,
we can improve upon current technology. For instance,
printer performance can be benchmarked and measured
using many advanced manufacturing software packages
that work in tandem with micro CT. Having better
feedback on print fidelity may enable assessment of 3D
bioprinting and encourage standardization of materials
performance. Even without tomographic imaging,
benchmark 3D prints such as the “3D Benchy” (http://
3DBenchy.com) have given the maker community of
open-source 3D printer operators a qualitative
Current Opinion in Biomedical Engineering 2017, 1:31–37
performance standard based on rendering of surface detail
and overhanging geometric features. Open-source bio-
printing, such as that utilized by Miller et al. and Hinton
et al., rely on the efforts of the RepRap movement (http://
reprap.org), including software capable of generating the
toolpaths (G-code) used by the majority of desktop 3D
printers [19,23,33]. These low-cost printers are built on
open-source frameworks where improvements can be
shared to benefit the community. The accessibility of
these 3D bioprinting platforms should encourage wider
adoption and foster further innovation. This should also
lead to standardized 3D metrology for assessing material
and printer performance, which will be required to tran-
sition 3D bioprinting into a clinically relevant bio-
fabrication platform.
Acknowledgements
This research did not receive any specific grant from funding agencies in
the public, commercial, or not-for-profit sectors.
References
Papers of particular interest, published within the period of review,
have been highlighted as:
* of special interest
* * of outstanding interest
1. Kinstlinger IS, Bastian A, Paulsen SJ, Hwang DH, Ta AH,
Yalacki DR, Schmidt T, Miller JS: Open-source selective laser
sintering (OpenSLS) of nylon and biocompatible poly-
caprolactone. PLoS One 2016, 11:e0147399.
2. Dubbin K, Hori Y, Lewis KK, Heilshorn SC: Dual-stage cross-
linking of a gel-phase bioink improves cell viability and ho-
mogeneity for 3D bioprinting. Adv. Healthc. Mater. 2016, 5:
2488–2492.
3. Hinton TJ, Hudson A, Pusch K, Lee A, Feinberg AW: 3D printing
PDMS elastomer in a hydrophilic support bath via freeform
reversible embedding. ACS Biomater. Sci. Eng. 2016, 2:
1781–1786.
4. Rengier F, Mehndiratta A, von Tengg-Kobligk H, Zechmann CM,
Unterhinninghofen R, Kauczor HU, Giesel FL: 3D printing based
on imaging data: review of medical applications. Int. J.
Comput. Assist. Radiol. Surg. 2010, 5:335–341.
5. Shah AM, Jung H, Skirboll S: Materials used in cranioplasty: a
history and analysis. Neurosurg. Focus 2014, 36:E19.
6. Zopf DA, Hollister SJ, Nelson ME, Ohye RG, Green GE: Bio-
resorbable airway splint created with a three-dimensional
printer. N. Engl J. Med. 2013, 368:2043–2045.
7. Mannoor MS, Jiang Z, James T, Kong YL, Malatesta KA,
Soboyejo WO, Verma N, Gracias DH, McAlpine MC: 3D printed
bionic ears. Nano Lett. 2013, 13:2634–2639.
8. Rhee S, Puetzer JL, Mason BN, Reinhart-King CA, Bonassar LJ:
3D bioprinting of spatially heterogeneous collagen con-
structs for cartilage tissue engineering. ACS Biomater. Sci.
Eng. 2016, 2:1800–1805.
9. Kang HW, Lee SJ, Ko IK, Kengla C, Yoo JJ, Atala A: A 3D bio-
** printing system to produce human-scale tissue constructs
with structural integrity. Nat. Biotechnol. 2016, 34:312–319.
The authors demonstrate the ability to utilize extrusion-based bio-
printing for fabrication of large anatomically-accurate constructs that
can be implanted into animals and studied for construct integrity and
tissue growth.
10. Daly AC, Cunniffe GM, Sathy BN, Jeon O, Alsberg E, Kelly DJ: 3D
bioprinting of developmentally inspired templates for whole
bone organ engineering. Adv. Healthc. Mater. 2016, 5:
2353–2362.
www.sciencedirect.com

11. Hung BP, Naved BA, Nyberg EL, Dias M, Holmes CA,
Elisseeff JH, Dorafshar AH, Grayson WL: Three-dimensional
printing of bone extracellular matrix for craniofacial regen-
eration. ACS Biomater. Sci. Eng. 2016, 2:1806–1816.
12. Chahal D, Ahmadi A, Cheung KC: Improving piezoelectric cell
printing accuracy and reliability through neutral buoyancy of
suspensions. Biotechnol. Bioeng. 2012, 109:2932–2940.
13. Miller JS: The billion cell construct: will three-dimensional
printing get us there? PLoS Biol. 2014, 12:e1001882.
14. Pati F, Jang J, Ha DH, Won Kim S, Rhie JW, Shim JH, Kim DH,
Cho DW: Printing three-dimensional tissue analogues with
decellularized extracellular matrix bioink. Nat. Commun. 2014,
5:3935.
15. Khademhosseini A, Langer R: A decade of progress in tissue
engineering. Nat. Protoc. 2016, 11:1775–1781.
16. Kolesky DB, Truby RL, Gladman AS, Busbee TA, Homan KA,
Lewis JA: 3D bioprinting of vascularized, heterogeneous cell-
laden tissue constructs. Adv. Mater. 2014, 26:3124–3130.
17. Kolesky DB, Homan KA, Skylar-Scott MA, Lewis JA: Three-
* dimensional bioprinting of thick vascularized tissues. Proc.
Natl. Acad. Sci. U. S. A 2016, 113:3179–3184.
This study showed that vascular networks can be printed using a
sacrificial ink within a thick cellularized tissue, and endothelialization of
the vascular channels support tissue viability and function.
18. Homan KA, Kolesky DB, Skylar-Scott MA, Herrmann J, Obuobi H,
Moisan A, Lewis JA: Bioprinting of 3D convoluted renal
proximal tubules on perfusable chips. Sci. Rep. 2016, 6:34845.
19. Miller JS, Stevens KR, Yang MT, Baker BM, Nguyen D-HT,
Cohen DM, Toro E, Chen AA, Galie PA, Yu X, et al.: Rapid
casting of patterned vascular networks for perfusable engi-
neered three-dimensional tissues. Nat. Mater. 2012, 11:
768–774.
20. Itoh M, Nakayama K, Noguchi R, Kamohara K, Furukawa K,
Uchihashi K, Toda S, Oyama J-i, Node K, Morita S: Scaffold-free
tubular tissues created by a bio-3D printer undergo remod-
eling and endothelialization when implanted in rat aortae.
PLoS One 2015, 10:e0136681.
21. Compaan AM, Christensen K, Huang Y: Inkjet bioprinting of 3D
silk fibroin cellular constructs using sacrificial alginate. ACS
Biomater. Sci. Eng. 2016.
22. Duarte Campos DF, Blaeser A, Buellesbach K, Sen KS, Xun W,
Tillmann W, Fischer H: Bioprinting organotypic hydrogels with
improved mesenchymal stem cell remodeling and minerali-
zation properties for bone tissue engineering. Adv. Healthc.
Mater. 2016, 5:1336–1345.
www.sciencedirect.com
3D bioprinting from the micrometer to millimeter Hinton et al. 37
23. Hinton TJ, Jallerat Q, Palchesko RN, Park JH, Grodzicki MS,
* * Shue H-J, Ramadan MH, Hudson AR, Feinberg AW: Three-
dimensional printing of complex biological structures by
freeform reversible embedding of suspended hydrogels. Sci.
Adv. 2015, 1.
The technique Freeform Reversible Embedded printing of Supported
Hydrogels developed in this study allows deposition of gelling hydro-
gels such as collagen I into a support slurry for fabrication of complex
geometries such as an embryonic chick heart.
24. Bhattacharjee T, Zehnder SM, Rowe KG, Jain S, Nixon RM,
Sawyer WG, Angelini TE: Writing in the granular gel medium.
Sci. Adv. 2015, 1.
25. Bhattacharjee T, Gil CJ, Marshall SL, Urueña JM, O’Bryan CS,
Carstens M, Keselowsky B, Palmer GD, Ghivizzani S, Gibbs CP,
et al.: Liquid-like solids support cells in 3D. ACS Biomater. Sci.
Eng. 2016, 2:1787–1795.
26. Ma X, Qu X, Zhu W, Li Y-S, Yuan S, Zhang H, Liu J, Wang P,
Lai CSE, Zanella F, et al.: Deterministically patterned bio-
mimetic human iPSC-derived hepatic model via rapid 3D
bioprinting. Proc. Nat.l Acad. Sci. U. S. A 2016, 113:2206–2211.
27. Claeyssens F, Hasan EA, Gaidukeviciute A, Achilleos DS,
Ranella A, Reinhardt C, Ovsianikov A, Shizhou X, Fotakis C,
Vamvakaki M, et al.: Three-dimensional biodegradable struc-
tures fabricated by two-photon polymerization. Langmuir
2009, 25:3219–3223.
28. Zhang W, Chen S: Femtosecond laser nanofabrication of
hydrogel biomaterial. MRS Bull. 2011, 36:1028–1033.
29. Ma Z, Koo S, Finnegan MA, Loskill P, Huebsch N, Marks NC,
Conklin BR, Grigoropoulos CP, Healy KE: Three-dimensional
filamentous human diseased cardiac tissue model. Bio-
materials 2014, 35:1367–1377.
30. Hahn MS, Miller JS, West JL: Three-dimensional biochemical
and biomechanical patterning of hydrogels for guiding cell
behavior. Adv. Mater. 2006, 18:2679–2684.
31. Wylie RG, Ahsan S, Aizawa Y, Maxwell KL, Morshead CM,
Shoichet MS: Spatially controlled simultaneous patterning of
multiple growth factors in three-dimensional hydrogels. Nat.
Mater. 2011, 10:799–806.
32. Tumbleston JR, Shirvanyants D, Ermoshkin N, Janusziewicz R,
Johnson AR, Kelly D, Chen K, Pinschmidt R, Rolland JP,
Ermoshkin A, et al.: Continuous liquid interface production of
3D objects. Science 2015, 347:1349.
33. Jones R, Haufe P, Sells E, Iravani P, Olliver V, Palmer C,
Bowyer A: RepRap – the replicating rapid prototyper. Robotica
2011, 29:177–191.
Current Opinion in Biomedical Engineering 2017, 1:31–37