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Vol.22 No.7 July 2004

Scaffold-based tissue engineering:

rationale for computer-aided design
and solid free-form fabrication systems
Dietmar W. Hutmacher1, Michael Sittinger2 and Makarand V. Risbud3
1

Division of Bioengineering and Department of Orthopaedic Surgery, National University of Singapore, 119260, Singapore
German Rheumatism Research Center and Experimental Rheumatology and Tissue Engineering Laboratory,
Department of Rheumatology and Clinical Immunology, Charite, Humboldt University of Berlin, 10117 Berlin, Germany
3
Graduate Program in Cell and Tissue Engineering and Department of Orthopaedic Surgery, Thomas Jefferson University,
Philadelphia 19107, USA
2

One of the milestones in tissue engineering has been

the development of 3D scaffolds that guide cells to form
functional tissue. Recently, mouldless manufacturing
techniques, known as solid free-form fabrication (SFF),
or rapid prototyping, have been successfully used to
fabricate complex scaffolds. Similarly, to achieve simultaneous addition of cells during the scaffold fabrication,
novel robotic assembly and automated 3D cell encapsulation techniques are being developed. As a result of
these technologies, tissue-engineered constructs can
be prepared that contain a controlled spatial distribution of cells and growth factors, as well as engineered
gradients of scaffold materials with a predicted microstructure. Here, we review the application, advancement and future directions of SFF techniques in the
design and creation of scaffolds for use in clinically
driven tissue engineering.
Currently, scaffold-based tissue engineering strategies are
expanding to encompass cells, bioactive molecules and
structural matrices. Each of these components is combined
into a construct that promotes repair and in the best
case scenario regeneration of damaged or diseased
tissues. Because many scaffold-based tissue-engineering
approaches are still experimental, it is not yet clear what
defines a so-called ideal scaffold.
Factors governing scaffold design are complex and
include considerations of matrix architecture, pore size
and morphology, mechanics versus porosity, surface
properties and degradation products. Moreover, because
scaffolds are often composite structures, other confounding variables include the composition of biological components and variation of these and other factors with time.
It could be argued that there is no ideal scaffold design
per se, instead each tissue requires a specific matrix design
with defined material properties. Scaffold design should
therefore begin with at least a set of minimum biochemical
and physical requirements [1]. The scaffold must provide
sufficient mechanical strength and stiffness to substitute
initially for wound contraction forces, and later for the
Corresponding author: Dietmar W. Hutmacher (biedwh@nus.edu.sg).

remodelling of the tissue. Furthermore, scaffold architecture should enhance initial cell attachment and subsequent migration into the matrix; it must also enhance
the mass transfer of metabolites and provide sufficient
space for remodelling of the organized tissue matrix and
development of a vasculature. To achieve this goal, the
scaffold degradation profile should be designed so that it
supports the construct until neotissue (cells plus organized
extracellular matrix without vascularization) is formed
[1]. A second important consideration is that if the
degradation profile is slow the 3D matrix will maintain
structural integrity and mechanical properties during the
in vitro and/or in vivo remodelling process. Factors
affecting the rate of remodelling include the type of tissue
and the anatomy and physiology of the host tissue [2]. The
external size and shape of the construct must also be
considered, particularly if the scaffold or cell construct is
customised for an individual patient. It is also imperative
that scaffolds are manufactured in a reproducible, controlled, and cost-effective fashion with the flexibility to
accommodate the presence of biological components, such
as cells and growth factors, in certain applications [3,4]. To
date, two methods of incorporating cells into scaffolds are
being explored: (i) seeding of cells onto the surface of the
scaffold following fabrication and (ii) the incorporation of
cells into the scaffold fabrication process. In terms of organ
printing, the second process is of considerable interest.
However, early results reflect success with printing cell
monolayers and not a complete 3D tissue or organ [5].
Basic considerations for scaffold fabrication by SFF
Advanced mouldless manufacturing techniques, commonly known as solid free-form fabrication (SFF), rapid
prototyping (RP) or, more colloquially, art to part technology [6] have recently been used for fabricating complex
shaped scaffolds. Unlike conventional machining, which
involves constant removal of materials, SFF builds parts
by selectively adding materials, layer by layer, as specified
by a computer program. Each layer represents the shape of
the cross-section of the model at a specific level. Today, SFF
is viewed as an efficient way of reproducibly generating
scaffolds of desired properties on a large scale [4,7,8]. In

www.sciencedirect.com 0167-7799/$ - see front matter q 2004 Elsevier Ltd. All rights reserved. doi:10.1016/j.tibtech.2004.05.005

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addition, one of the potential benefits of SFF technology is

the ability to create parts with highly reproducible
architecture and compositional variation.
Over the past two decades, .20 RP systems were
developed and commercialised [9] with a focus on the rapid
manufacturing of prototypes for non-biomedical applications. Very recently, biomaterial scientists used these
technologies to fabricate scaffolds for tissue engineering.
SFF techniques offer unique ways to precisely control
matrix architecture (size, shape, interconnectivity, branching, geometry and orientation) yielding biomimetic structures varying in design and material composition, thereby
enhancing control over mechanical properties, biological
effects and degradation kinetics of the scaffolds. RP
techniques can be easily automated and integrated with
imaging techniques to produce scaffolds that are customised in size and shape allowing tissue-engineered grafts to
be tailored for specific applications or even for individual
patients (Figure 1).
A recent milestone in scaffold fabrication by SFF was
the development of solvent-free and aqueous-based systems. By allowing inclusion of bioactive components, such
as growth factors, cells and drugs, these systems offer new
opportunities in tissue engineering and regenerative
medicine. In the following sections we discuss the state
of the art in SFF techniques applied to scaffold fabrication.
We have classified the SFF systems based on their
processing technology. Future directions in the design
and manufacturing of novel and patient-specific matrices
(a)

for clinically relevant bone-engineering applications will

also be discussed.
SFF techniques used in scaffold fabrication
Systems based on laser technology
Stereolithography apparatus. Stereolithography (SLA) is
often considered the pioneering RP technique with the
first commercial system introduced in 1988 by 3D Systems
Inc. (www.3dsystems.com). This technique is based on the
use of a UV laser that is vector scanned over the top of a
bath of a photopolymerisable liquid polymer material. As
polymerisation is initiated, the laser beam creates a first
solid plastic layer, at, and just below the surface of the
bath. This laser polymerisation process is repeated to
generate subsequent layers by tracing the laser beam
along the design boundaries and filling in the 2D crosssection of the model, layer-by-layer. Once the model is
complete, the platform is raised out of the vat and the
excess resin is drained. The model is cured in a UV oven
and finished by smoothing the surface irregularities.
Resolution of a standard SLA layer is determined by the
elevator layer resolution (up to 1.3 mm) and laser spot size
(80 250 mm) [10].
The use of SLA in the biomedical industry is currently
limited to the creation of anatomical models for surgical
planning or teaching [11,12]. Because of curing and
shrinkage after post-processing a shortfall of the SLA
model is compromised resolution [13]. Also, due to absorption and scattering of the laser beam, a pronounced

(b)

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(e)

(f)

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Figure 1. Tissue engineering of patient-specific bone grafts. This innovative treatment protocol uses medical imaging, computational modelling and bioresorbable scaffold
fabricated with rapid prototyping (RP) technique. CT scan data of the patients bone defect (a) are used to generate a computer-based 3D model (b). This model is then
imported into RP system software to be sliced into thin horizontal layers, with the tool path specified for each layer (c). The sliced data are used to instruct the RP machine
(d) to build a scaffold (e) layer by layer, based on the actual shape of the computer model (c). RP technology produces excellent templates for the treatment of intricate
bone defects (a and f). Custom-made scaffold and cell constructs (g, see arrows) exactly follow the complex shaped 3D contour of the skull.
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deformation occurs when smaller and more intricate

objects are fabricated. Generally, the manufactured part is
weak at the time of removal and needs post-processing for
further curing [10,13]. It is noteworthy that there is a
limited choice of photopolymerisable biomaterials that
have the required biodegradability and biocompatability,
mechanical stability and other prerequisite properties for
scaffold applications. Examples of photopolymerisable
macromers include derivatives of polyethylene glycol
(PEG) acrylate, PEG methacrylate, polyvinyl alcohol
(PVA) and modified polysaccharides such as hyaluronic
acid and dextran methacrylate. Polypropylene fumarate,
anhydride and polyethylene oxide (PEO) precursor systems could be explored because investigations are underway for their clinical use as curable bioadhesives or
injectables. Cooke et al. recently demonstrated the
feasibility of using the SLA process to build and control
3D multilayer parts made from a biodegradable, biocompatible resin polypropylene fumarate [14]. Likewise,
Matsuda and Mizutani have developed a biodegradable,
photocurable copolymer, acrylated capped poly-1-caprolactone-co-trimethylene carbonate to be used with the SLA
apparatus to prepare photoconstructs such as microneedles, a microcylinder and microbanks on surfaces
[15]. Based on such encouraging results micro stereolithography (mSL) in particular offers great potential for the
production of 3D polymeric structures with micrometer
resolution. Recently, mSL was also tested for fabricating
high aspect ratio and complex 3D microstructures [16,17].
Selective laser sintering. The selective laser sintering
(SLS) technique uses a CO2 laser beam to sinter thin layers
of powdered polymeric materials, forming solid 3D objects.
During SLS fabrication, the laser beam is selectively
scanned over the powder surface following the crosssectional profiles carried by the slice data. The interaction
of the laser beam with the powder raises the powder
temperature and sintering occurs at just beyond the glass
transition temperature causing the particles to fuse
together to form a solid mass. Subsequent layers are
built directly on top of previously sintered layers with new
layers of powder being deposited by a roller [18]. Rimell
and Marquis have reported the fabrication of clinical
implants using a simplified selective laser sintering
apparatus and ultra high molecular weight polyethylene
(UHMWPE) [19]. It was observed that solid linear continuous bodies could be fabricated, but material shrinkage
occurred when a sheet-like structure was desired. The
porosity of the material formed was also a concern. The
material exposed to the laser beam was seen to undergo
degradation in terms of chain scission, cross-linking and
oxidation. It was concluded that to apply this technology to
the fabrication of UHMWPE devices requires the development of improved starting powders with increased density.
Lee and co-workers were the first to use SLS to manufacture ceramic bone implants [20]. The augmentation of
alveolar ridge defects in canines was conducted to assess
the safety and efficacy of these SLS-fabricated calcium
phosphate (CaP) implants [21]. Histological evaluation
revealed that the implant material was biocompatible and
mineralized bone was formed in the macro pores. Griffith
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Vol.22 No.7 July 2004

and Halloran have reported the fabrication of ceramic

parts using suspensions of alumina, silicon nitride and
silica particles in a UV photocurable monomer by SLA [22].
Using a similar technique, a suspension of hydroxyapatite
(HA) in a photocurable monomer was formulated to
produce scaffolds for orbital floor prosthesis [23]. It was
concluded that for bone graft applications HA scaffolds
provide a superior cosmetic appearance compared with
conventional techniques [23]. Porter et al. formulated
suspensions of calcium polyphosphate (CPP) and a photocurable monomer for forming bioresorbable skeletal
implants using SLA [24]. Sintering CPP at 6008C for 1 h
produced a crystalline material (average porosity of 22.9%)
exhibiting superior bend strength and toughness compared with amorphous CPP. Tan et al. used different
compositions of non-degradable polyetheretherketone
(PEEK)/HA powder blends to assess their suitability for
SLS processing. [25].
System based on print technology 3D printing. 3D
printing (3DP) technology was developed at MIT [3] and
became one of the most investigated SFF techniques
in tissue engineering and drug-delivery applications
[26 31]. An advantage of 3DP is that it can be performed
in an ambient environment. The 3D printer constructs the
3D model by first spreading a layer of fresh powder over a
building platform. An inkjet print head prints or deposits
the binder solution onto the powder bed. After the 2D layer
profile is printed, a fresh layer of powder is laid down. The
printing cycle continues and the layers merge together
when fresh binder is deposited until the whole part is
completed. After the binder has dried in the powder bed,
the finished component is retrieved and unbound powder
removed. However, if the part is designed to be porous, one
drawback of a powder-supported and powder-filled structure is the difficulty in removing internal unbound powder.
The resolution of the 3D printer is also limited by the
nozzle size and the degree of control allowed over the
position controller that defines print head movement. In
addition, the particle size of the powder governs the
layer thickness (usually 80 250 mm). In the context of
tissue-engineering scaffolds, base biomaterials are not
usually available in powder form and require special preprocessing for 3DP [26 28]. The surface roughness and
aggregation properties of the powdered materials also
affect the component resolution and efficiency of removal of
trapped materials [29,31].
Work by Lam et al. demonstrated the feasibility of
using natural biopolymers (starch, dextran and gelatin)
and distilled water as the binder [31]. This aqueous system
eliminated the problem linked to the use of an organic
solvent. However, the scaffold is bound by water and is
therefore water-soluble, necessitating a lengthy postprocessing step to waterproof the product. This work
also showed that owing to non-elimination of all the microspaces and gaps present between the fused powdered
particles, scaffolds manufactured by 3DP possessed a
second tier of porosity referred to as microporosity. This
microporosity is probably created at random and could be
integral in the degradation kinetics of the scaffold.

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Assembly technology-based systems

Shape deposition manufacturing. The term shape deposition manufacturing (SDM) was initially introduced for
the fabrication of scaffolds for bone tissue engineering [32].
It involves the fabrication of a layered scaffold in a
customised geometry by processing the clinical imaging
data and translating it to the desired scaffold layer (,1 mm)
by a computer-numerically-controlled cutting machine
(http://www-2.cs.cmu.edu/People/tissue/front_page.html).
This technique employed the simultaneous addition of
cells to the matrix during 3D scaffold fabrication. In a
published study, the scaffolds were incrementally built up
from prefabricated cross-sectional layers of foams [,1 mm
thick, made from blends of polycaprolactone (PCL) and
poly D,L lactide-co-polyglycolide (P[D]LGA) combined
with HA granules]. Layers were manually stacked up
and joined to form 3D discs with biodegradable or nonbiodegradable fasteners. Each prefabricated layer was
first seeded with cells and growth factors before final
assembly. Although unique, this technique is dependent on
the quality of the fabricated foam, which is generated by a
conventional method. Similarly, because this technique is
still in its infancy, assembly is not automated. Furthermore, to avoid trapping of seeded cells within each layer,
the assembly process needs to be controlled so that there is
sufficient pore interconnectivity.
The concept of a novel robotic micro-assembly technique
was first published in 2000 [1] and recently the first
prototype system was developed and fabricated and
the parts were successfully made [33,34]. The principle
of micro-assembling a functional tissue-engineering
scaffold is based on the same concept as assembling a
structure using small building-block units like Legow.
Building blocks of different designs are first fabricated
using lithography, or other microfabrication technologies.
Then the blocks are assembled by a computer-controlled,
dedicated precision robot with four degrees of freedom
and micro-gripping capabilities (Figure 2). The final
result is a tissue graft built out of cells and different
materials with the required biochemical and physical
properties.
The application of advanced microfabrication techniques such as silicon micro-machining and polymer
replica moulding for tissue engineering of complex tissues
has been described [35,36]. The goal of this work was to
produce organ templates having feature resolution of 1m,
well in excess of that necessary to fashion the capillaries
comprising the microcirculation of the organ. Initial efforts
have resulted in high-resolution polymer scaffolds produced by replica moulding from silicon micro-machined
template wafers. These scaffolds have been successfully
seeded with endothelial cells in channels with dimensions
as small as the blood capillaries [35,36].
Extrusion technology-based systems. Techniques such as
fused deposition modelling (FDM), 3D plotting, multiphase jet solidification (MJS) and precise extrusion
manufacturing (PEM) employ extrusion of a material in
a layered fashion to build a scaffold. Depending on the type
of machine, a variety of biomaterials can be used for
scaffold fabrication.
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A traditional FDM machine consists of a head-heated

liquefier attached to a carriage moving in the horizontal xy
plane [37]. The function of the liquefier assembly is to heat
and pump the filament material through a nozzle directly
on to the build platform following a programmed path.
Once a layer is built, the platform moves down one step in
the z direction to deposit the next layer. Parts are made
layer by layer with the layer thickness varying in
proportion to the nozzle diameter. FDM is restricted to
the use of thermoplastic materials with good melt viscosity
properties; cells cannot be encapsulated into the scaffolds
during the fabrication process.
An interdisciplinary group in Singapore has studied
and patented the parameters for processing PCL and
several composites (PCL/HA, PCL/TCP etc.) by FDM [38].
These first generation scaffolds (PCL) have been studied
for more than three years with, and without, cells in
a clinical setting (http://www.osteopore-intl.com). The
second generation scaffolds for bone engineering using
FDM were made of polymer and CaP composites because
they confer favourable mechanical and biochemical
properties, including strength via the ceramic phase,
toughness and plasticity via the polymer phase, favourable
degradation and resorption kinetics and graded mechanical stiffness. Other advantages include improvement of
cell seeding, and the enhanced incorporation and immobilization of growth factors. Endres et al. [39] and Rai et al.
[40] have tested these PCL/CaP composite scaffolds for
bone engineering and reported encouraging results
(Figure 3). Woodfield et al. used a FDM-like technique
for producing scaffolds made of polyethylene glycolterephthalate-polybutylene terepthalate (PEGT/PBT)
[41,42]. By varying PEGT/PBT composition, porosity and
pore geometry, scaffolds were produced with a range of
mechanical properties for engineering of articular
cartilage.
A variation of the FDM process, the precision extruding
deposition (PED) system, was developed and tested at
Drexel University [43]. The major difference between PED
and conventional FDM is that the scaffolding material can
be directly deposited without filament preparation. Pelletformed PCL is fused by a liquefier temperature provided
by two heating bands and respective thermal couples and
is then extruded by pressure created by a turning precision
screw.
Several groups [44 51] and companies (Envisiontec;
http://www.envisiontec.de, Sciperio; http://www.Sciperio.
com) have developed SFF machines that can perform
extrusion of strands or filaments and/or plotting of dots in
3D (Figure 4). These systems are built to make use of a
wide variety of polymer hotmelts as well as pastes or
slurries, solutions and dispersions of polymers and
reactive oligomers. One such technique, MJS, involves
the extrusion of a melted material through a nozzle, which
has a jet-like design. The MJS process is usually used to
produce metallic or ceramic parts via a lost-wax method.
Poly (D,L)-lactide structures for bone and cartilage tissue
engineering have been fabricated using a MJS machine
built by the Frauenhofer Institue, Stuttgart [44]. The
scaffolds had a pore size of 300 400 mm and supported
ingrowth of human bone tissues. Calvert et al. built an

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(b)

(a)
Articular
cartilage

60m
500m

Defect
Bone
200m
Introduce bone cells
from the patient

90m
Biometric
surface

Scaffold

60m

(c)

Grow into an artificial bone

Transplantation

(d)

(e)

Visual feedback
through a microscope

Micro-gripper
60m
Haptic interface

y
x
x-y-z table
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Figure 2. Hutmachers group is developing a novel method based on robotic micro-assembly to fabricate scaffold and cell constructs for a variety of tissue-engineering
applications (a). The concept is based on the assembly of microscopic Legow-like building blocks into a scaffold (b,c). The computer-controlled and automated system
allows the control of distribution of growth factors and living cells within the scaffold in 3D (d) so that scaffold and cell constructs with customised biological and physical
properties can be developed. The micro-assembly is carried out using an in-house designed precision robot. u represents the angle that the gripper can rotate round its
own axis (3608) A two-finger microgripper (e, arrow) was developed to grasp, move and assemble the microparts. Building blocks were fabricated using micro stereolithography (mSLA) and a micro-electro-mechanical systems (MEMS) technique (c), which has the capability of mass production of micro-objects with complex shapes and high
precision at an economical cost [33,34].

in-house extrusion-based system that could fabricate

scaffolds with a resolution of , 0.5 mm and typical layer
heights of 0.2 1.0 mm [45 47]. Xiong et al. have developed a RP machine termed PEM and fabricated porous
poly L-lactic acid (PLLA) with tricalcium phosphate (TCP)
scaffolds for bone tissue engineering [48].
Landers et al. [49] used the term bioplotter to describe
the fabrication of scaffolds of different composition.
Versatility of this technique was demonstrated in several
other studies [50 52]. A study compared 3DP and 3D
bioplotting for the manufacture of biodegradable polyurethane scaffolds using aliphatic polyurethanes based on
lysine ethyl ester diisocyanate and isophorone diisocyanate [53]. Layer-by-layer construction of the scaffolds
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was performed by 3DP (i.e. bonding together starch

particles followed by infiltration and partial crosslinking
of starch with lysine ethyl ester diisocyanate). Alternatively, the 3D bioplotting process permitted 3D dispensing
and reactive processing of oligoetherurethanes derived
from isophorone diisocyanate, oligoethylene oxide, and
glycerol.
A rapid prototyping robotic dispensing (RPBOD)
system that works on the same principles as the 3D
bioplotter was described by Ang et al. [54]. These
investigators produced 3D chitosan and chitosan HA
scaffolds using the RPBOD. For this purpose, solutions of
chitosan or chitosan HA were extruded into a sodium
hydroxide and ethanol bath. It was noted that the

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(b)

(f)
(c)

(d)

(e)

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Figure 3. (a-f) In vitro studies of polycaprolactone calcium phosphate (PCL/CaP) composites made by fused deposition modeling (FDM) showed that bone-marrow-derived
precursor cells are able to attach, migrate, proliferate and differentiate. Cells are able to span across the large pores and pore interconnections (a, F-actin staining, 100
magnification) and produce mineralised extracellular matrix (b, von Kossa staining, 100 magnification). Scanning electron micrographs (c) reveal that after 3 weeks the
entire scaffold architecture is filled with neotissue. In preliminary in vivo studies the PCL/CaP scaffolds showed good tissue integration as well as a minimal foreign body
reaction in the subcutaneous (d) as well as intramuscular (e) tissue when implanted into rabbits. MicroCT analysis (courtesy, Dr. Robert Guldberg, Georgia Institute of Technology) of a second generation FDM scaffold for bone engineering shows the homogenous distribution of the CaP particles in the PCL matrix and on the scaffold surface (f).

concentration of sodium hydroxide controlled the adhesion

between layers. In another study, Vozzi et al. have
achieved resolution as low as 10 mm on a 2D structure
through the use of a system built by a computer-controlled,
three-axis micropositioner, microsyringes and electronically regulated air pressure valves [55,56].
The advantage of all these extrusion systems is their
versatility. However, current encapsulation [57] and organ
printing techniques [4] that use this SFF technique are
restricted to the mm scale and might not yet permit
fabrication of a true 3D tissue construct.
Solid ground curing. The wider application of solid ground
curing (SGC) in designing scaffolds is mainly driven by
developments of photochemically driven gelation technology of biomacromolecules that are chemically modified
with photodimerizable groups. Recent reviews summarize
the chemistry and rationale for using these polymers in
scaffold-based tissue engineering [58,59]. Photopolymerizable and biodegradable polyethyleneglycol-based macromers, acrylated polyethyleneglycol derivatives including
polyethylene glycol-co-polyhydroxy acid diacrylate and
polyethylene glycol-polylysine diacrylate, both of which
are end-capped with acryloyl groups, have been studied in
detail by Matsudas group [60].
Using SCG technology, tubular photoconstructs were
prepared by photocopolymerization of vinylated polysaccharide and vinylated gelatin [61]. The mixing of diacrylated polyethylene glycol with vinylated polysaccharide
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improved the burst strength of photogels against the

gradual infusion of water. These photocurable polysaccharides might be used as photocured scaffolds in
tissue-engineered devices [61].
Liu and Bhatia describe the development of a photopatterning technique that allows localized photoencapsulation of live mammalian cells to control the tissue
architecture. Cell viability was characterized using
HepG2 cells, a human hepatoma cell line. The utility of
this method was demonstrated by photopatterning hydrogels containing live cells in various single layer structures,
patterns of multiple cellular domains in a single hybrid
hydrogel layer, and patterns of multiple cell types in
multiple layers. The authors observed that UV exposure
itself did not cause cell death at the doses and time
scale studied, although the photoinitiator 2,2-dimethoxy2-phenyl-acetophenone was itself cytotoxic in a dosedependent manner. Furthermore, the combination of UV
and photoinitiator was the least biocompatible condition
presumably due to formation of toxic free radicals. [62].
Indirect SFF in scaffold fabrication
A key factor used to enhance the versatility of scaffold
fabrication by RP or SFF is construction of a scaffold
matrix using a wide variety of biomaterials. One emerging
method, which adds another level of versatility to SFF, is
to fabricate a negative mould based on the scaffold design
and cast the scaffold using desired polymeric and/or
ceramic biomaterials. This alternative technique is termed

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(a)

(b)

z axis
z axis

Rotation about z axis

(c)

(d)

(e)

(f)

(g)

(h)

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Figure 4. A group at the National University of Singapore designed and built a

rapid prototyping (RP) machine (a-d) and software, which is specifically dedicated
to scaffold fabrication. The machine is based on an extrusion dispenser head (multiple heads are also possible) by a 3-axis robot (a). The process generates a scaffold from a computer file (STL etc.) by building micro strands or dots. Depending
on the machine set up a tissue engineer can make use of a wide variety of polymer
pastes and solutions (b, chitosan), hot melts (e-h) as well as and dispersions and
chemical reactive systems (e.g. fibrin glue). Recently, the group processed and studied several novel di- and tri-block copolymers (Hutmacher et al., unpublished
data). PEG-PCL, PCL-PEG-PCL, PCL-PLA-PEG and PCL-urethane-PEG scaffolds with
a 4 (e, g) and 6 (f, h) angle pattern were studied via confocal laser microscopy (g, h)
using a life (2 mg/mL fluorescein diacetate FDA, green) death (0.1 mg/mL propidium iodide PtdIns, red) assay. Abbreviations: PCL-PEG-PCL, poly(1-caprolactone)
(PCL)-poly(ethylene glycol)-Poly(1-caprolactone); PCL-PLA-PEG, poly(1-caprolactone) poly(DL-lactide) (PCL)-poly(ethylene glycol); PEG-PCL, poly(1-caprolactone)
(PCL)-poly(ethylene glycol); STL, stereolithography.

indirect SFF [63]. Chu et al. created HA scaffolds with

interconnecting pores based on a lost mould technique by
combining epoxy resin moulds made by SLA and computeraided design (CAD) data [64]. Despite a few fabrication
inaccuracies, in vivo experiments demonstrated osteoconductivity and biocompatibility of these scaffolds in a
minipig model. Taboas et al. have created a series of
biomimetic scaffolds by mould removal in combination
with conventional sponge fabrication [65]. The fabricated scaffolds had interconnected pores ranging from
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Vol.22 No.7 July 2004

500 800 mm as specified by the pre-fabricated mould,

and when local pores were formed they ranged from
10 300 mm depending on the local pore creating method.
Similarly, Wilson et al. [66] have developed an indirect SFF
method for ceramic scaffolds with defined and reproducible
3D porous architectures; they report ectopic bone formation for all scaffold and cell constructs.
Manufacturing of collagen-based scaffolds by using the
SFF technique to fabricate a mould has also been reported
[67]. The mould was dissolved away with ethanol and the
collagen scaffold was then critical-point dried with liquid
CO2. SLA models derived from x-ray computer tomography have been used to generate biocompatible and
biodegradable heart valve scaffolds from poly-4-hydroxybutyrate (P4HB) and polyhydroxyoctanoate (PHOH) by
thermal processing [68]. Use of thermoplastic elastomers,
P4HB and PHOH, allowed moulding of a complete
trileaflet heart valve scaffold without the need for suturing
or other post-processing, and demonstrated the ability of
indirect SFF to reproduce complex anatomical structures.
Taken together, these studies demonstrate that indirect
SFF adds a greater degree of versatility and detail to
scaffold design [69]. The previous restriction on casting
was the inability of moulds to produce complex geometry
and internal architecture but with indirect SFF conventional casting processes with these SFF moulds can meet
specific tissue-engineering requirements, including the
need for mechanical integrity and customised shapes. In
addition, indirect SFF allows use of a wider range of
biomaterials or a combination of materials (composites
and/or co-polymers). However, some drawbacks still exist
with this technology, including the use of organic solvents
and the resolution of the SFF method. For example, the
cast model inherits errors and defects from the mould,
such as cracks and dimensional changes. Also, a method
must be developed to remove the mould precisely while
preserving the cast scaffold intact without compromising
its properties.
Conclusions
In many ways, remodelling of a graft generated by scaffoldbased tissue engineering can be considered analogous to
guided wound healing in that most constructs become
extensively remodelled as part of the normal tissue-repair
process. Therefore, it might not be necessary to produce
very precise structures or replicas of biological tissues
(e.g. trabecular bone structures) when designing a scaffold. RP and SFF techniques provide CAD-supported
manufacturing processes that are capable of reproducibly
fabricating scaffolds from a variety of biomaterials with
different physical and biochemical properties. These
techniques offer the right balance of capability, practicality
and cost benefit that make them suitable for fabrication of
constructs in sufficient quantity and quality for clinically
driven tissue-engineering applications.
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