Materials Science and Engineering C 20 (2002) 49 – 56

www.elsevier.com/locate/msec

Scaffold development using 3D printing with a starch-based polymer

C.X.F. Lam a, X.M. Mo a, S.H. Teoh a, D.W. Hutmacher b,*
a
Laboratory for Biomedical Engineering, Division of Bioengineering, Department of Mechanical Engineering,
National University of Singapore, Singapore
b
Department of Orthopedic Surgery, National University of Singapore, Singapore

Abstract

Rapid prototyping (RP) techniques have been utilised by tissue engineers to produce three-dimensional (3D) porous scaffolds. RP
technologies allow the design and fabrication of complex scaffold geometries with a fully interconnected pore network. Three-dimensional
printing (3DP) technique was used to fabricate scaffolds with a novel micro- and macro-architecture. In this study, a unique blend of starch-
based polymer powders (cornstarch, dextran and gelatin) was developed for the 3DP process. Cylindrical scaffolds of five different designs
were fabricated and post-processed to enhance the mechanical and chemical properties. The scaffold properties were characterised by
scanning electron microscopy (SEM), differential scanning calorimetry (DSC), porosity analysis and compression tests. D 2002 Elsevier
Science B.V. All rights reserved.

Keywords: Three-dimensional printing; Rapid prototyping; Starch-based scaffolds; Tissue engineering

1. Introduction Helisys [2], fused deposition modeling (FDM) by Stratasys

Traditional methodologies of scaffold fabrication include
fiber bonding, solvent casting and particulate leaching,
membrane lamination, melt molding and gas foaming.
However, the drawbacks of these techniques include the
extensive use of highly toxic organic solvents, long fab-
rication periods, labour-intensive processes, incomplete
removal of residual particulates in the polymer matrix, poor
repeatability, irregularly shaped pores, insufficient intercon-
nectivity of pores and thin structures. In addition, most of
these methods bear restrictions on shape control. For exam-
ple, casting is dependent on the shape of the mould and not
on being able to easily and independently control the
porosity and pore size of the scaffolds [1].
Rapid prototyping (RP) is the process of creating three-
dimensional (3D) objects through repetitive deposition and
processing of material layers using computer-controlled
equipment. It is based on the 2D cross-sectional data
obtained from slicing a computer-aided design (CAD)
model of the object. Presently, there are numerous RP
technologies available including stereolithography appara-
tus (SLA) by 3D systems [2], selective laser sintering (SLS)
by DTM [2], laminated object manufacturing (LOM) by
*
Corresponding author. Tel.: +65-874-5105; fax: +65-777-3537.
E-mail address: mpedwh@nus.edu.sg (D.W. Hutmacher).
[1,2] and three-dimensional printing (3DP) by the Massa-
chusetts Institute of Technology (MIT) [1 –3]. As the scaf-
fold design is based on a computer software model, the
desired 3D interconnectivity and structure can be accurately
controlled by the highly repeatable computer-controlled
fabrication process.
The 3DP technology was developed at the Massachusetts
Institute of Technology (MIT) [3,4]. Basically, the 3DP is a
layered fabrication process, in which the sliced 2D profile of a
computer model is printed on a fresh layer of powder via
deposition of a suitable binder. Successive 2D profiles are
then printed on a freshly laid layer of powder until the whole
model is completed. The printed binder would join the
respective profiles of each layer together. The part is com-
pleted upon removal of the unbound powder and suitable
post-processing [5]. In the past, the technique was mainly
used to fabricate drug delivery devices [6]. Recently, this
capability of creating 3D structures attracted tissue engineers
to apply the technology to design and fabricate scaffolds.
The works of Kim et al. [7] have demonstrated the
potential of the 3DP technology when it is combined with
salt leaching technique in the fabrication of polymeric
scaffolds. The material used was copolymers of polylac-
tide – coglycolide (PLGA, 85L:15G) and a suitable solvent.
They fabricated cylindrical scaffolds (F 8
7 mm) and
managed to achieve interconnected porous channels of
about 800 Am and microporosities of 45– 150 Am by using

0928-4931/02/$ - see front matter D 2002 Elsevier Science B.V. All rights reserved.
PII: S 0 9 2 8 - 4 9 3 1 ( 0 2 ) 0 0 0 1 2 - 7
50 C.X.F. Lam et al. / Materials Science and Engineering C 20 (2002) 49–56
salt leaching. Their experiment on seeding hepatocytes on
these scaffolds yielded successful attachment of large num-
bers of hepatocytes on the scaffolds.
Zeltinger et al. [8] studied the cellular reactions to pore
size, and void fractions were investigated based on 3DP-
fabricated scaffolds. In the experiment, scaffolds of varying
pore sizes (38 – 150 Am) and void fractions (75% and 90%)
were fabricated using the 3DP technology and salt leaching
technique. Poly(L-lactide) (L-PLA) and chloroform were
used to construct the scaffolds, 10 mm in diameter and 2
mm in height. Cell proliferation was observed on the
scaffolds, but results varied between cell types and the
experimental parameters.
All the studies executed by the MIT groups used ali-
phatic polymers as the matrix material; hence, the reported
studies required the use of organic solvents. The aim of our
group is to explore the feasibility of using 3D printing in
combination with natural polymers and water-based binders.
2. Materials and methods
2.1. Materials
Recently, a 3D printer was developed and commercial-
ized by Zcorp (Z402), which uses natural polymers as well
as plaster of paris in combination with a water-based ink.
This commercial machine was used for the study. A suitable
biomaterial, which had to be sourced as the standard
material, and binder for the Z402 3D printer were meant
for non-tissue engineering purposes. The properties of this
biomaterial would require that it operate well within the
parameters and constraints of the printer. A three-powder
blend was formulated for the experiments. It consisted of 50
wt.% cornstarch (Max’s, Holland; f 10 Am), 30 wt.%
dextran (Sigma, USA; > 100 Am) and 20 wt.% gelatin (Nitta
Gelatin, Japan; >100 Am). Starch is a polysaccharide pro-
duced by higher plants as energy storage [9] and is composed
of two polymers of D-glucose: amylose and amylopectin.
Amylose is a lightly branched polymer, while amylopectin
is highly branched with an overall tree-like structure.
Dextran is a polysaccharide secreted by certain strains of
bacteria and slimes [9]. Gelatin is made from the naturally
occurring collagen in animals. It is a polypeptide and
contains 18 different amino acids. Distilled water was used
as the binder in combination with a blue dye to ease
processing and investigations.
2.2. Design and fabrication
In this study, scaffolds were designed using a CAD
software (Unigraphics v. 15.0.3). Four different porous
cylindrical scaffolds (F 12.5
12.5 mm) and a solid
cylinder that served as control were modeled. The pores
were either cylindrical ( F 2.5 mm) or rectangular
(2.5
2.5 mm). The designs were exported in a rapid
prototyping format (.stl file format) and uploaded to the
3DP software where the model was sliced by a slicing
algorithm. These 2D sliced layers were then built by the
printer layer by layer till the final object was physically
formed. The designs are shown in Table 1.
Post-processing was necessary to enhance the strength of
the scaffolds and also to increase its resistance against water
uptake. The scaffolds were dried at 100 jC for 1 h after
printing. The scaffolds were strong enough to be manually
handled and the unbound powders were removed without
damaging the integrity of the scaffolds. Two different
batches of scaffolds were prepared. The first batch was
infiltrated with different amounts of copolymer solution,
which was made by dissolving the copolymer of 75%
poly(L-lactide) acid and 25% polycaprolactone (Gunze,
Japan) in dichloromethane (CH2Cl2) (J.T. Baker), to form
a concentration of 2%. Groups L1, L2 and L3 were
infiltrated with 3.5, 5.2 and 6.9 ml, respectively. They were
left to evaporate and dry at room temperature (27 jC). The
second batch of scaffolds was infiltrated by the same
method, but after drying, the scaffolds were soaked in
deionised water for 10 min and dried at 100 jC.
2.3. Analysis and testing
2.3.1. Water absorption tests
The infiltrated scaffolds were soaked in water for 10 min
and evaluated for the amount of water absorbed. An exten-
sive 3-day water absorption test was also conducted.
2.3.2. Scanning electron microscopy (SEM)
Microstructural characterisation and particle size analysis
of the materials and the scaffolds were conducted using an
SEM (JEOL JSM-5800 LV). Uninfiltrated scaffolds, infil-
trated scaffolds before chemical bonding and infiltrated
scaffolds after chemical bonding were examined under high
vacuum conditions at 15 kV.
2.3.3. Differential scanning calorimetry (DSC)
Differential scanning calorimetry (DSC 2910, TA Instru-
ments) was used to analyse the thermal transition of the
powders, copolymer and the fabricated scaffolds. The three
Table 1
Fabricated scaffolds (overall dimensions: F 12.5
12.5 mm)
 C.X.F. Lam et al. / Materials Science and Engineering C 20 (2002) 49–56
samples studied were: the three-powder blend (powder only),
PLLA – PCL copolymer (polymer only) and the infiltrated
and soaked scaffold matrix. They were scanned from room
temperature (25 jC) up to 250 jC at a rate of 10 jC/min.
2.3.4. Mechanical tests
Compression tests were conducted with an Instron 4302
material testing system using a 1-kN load-cell (Canton, MA,
USA). Scaffolds were tested in accordance with ASTM
Standard D695-96 and were conducted under both dry and
wet conditions. The scaffolds were soaked for 4 h at 37 jC in
phosphate-buffered saline to simulate in vivo conditions.
Three sets of specimens were tested: uninfiltrated (dry),
infiltrated (dry) and infiltrated (wet). In each set, there were
five specimens of each design. Both the compressive stiffness
(E) and initial compressive yield strength were evaluated
from the stress – strain graph plotted from the load displace-
ment graph obtained from the compression tests (cross-sec-
tional area was determined from the measurement of the
scaffold dimensions, e.g. area for the ‘‘rod_censolid’’ scaffold
would be the area of four F 2.5-mm ‘‘rods’’ deducted from
the main circular scaffold surface area). Stiffness was eval-
uated from the initial ‘‘linear-elastic’’ region of the stress –
strain graph and initial yield strength was taken at the first
yield point on the stress – strain graph in MPa.
2.3.5. Porosity analysis
Macroporosity is achieved by the macropores created in
the design. For powdered materials, the microporosity is
created by space between the individual granules of powder.
Both porosities have to be considered and were calculated
by the general formula:
ðV a  V t Þ
Porosity, P ¼ , enzymatic and acid hydrolysis [16]. Although there was
Va
where Va = apparent volume and Vt = true volume.
Macroporosity was calculated from the actual dimen-
sions and volume of the scaffolds. Both the design porosity
(from CAD) and actual porosity were measured. Va was
taken to be the volume of the solid cylinder and Vt, the
measured or calculated volume. As for calculating the
microporosity, the Va referred to the Vt from the macro-
porosity measurement for each of the respective designs,
while Vt was measured using a gas pycnometer (Accupyc
1330, Micromeritics Instrument) in pure helium to deter-
mine the bulk volume of the scaffolds.
Table 2
Mass evaluation of the water absorption test for the various infiltration volumes
Coat Dry (g) First Water
10 min (g) absorbed (g)
L1 0.282 0.555 0.273
L3 0.309 0.532 0.223
51
3. Results and discussions
3.1. Effect of post-processing on scaffold integrity
Reis et al. [10] showed the potential application of starch
as a biomaterial. As individual components, dextran and
gelatin are biocompatible [11]; hence, a combination of
these three materials would be a useful biomaterial. Using
water as the binder means that the problem of a toxic
fabrication environment is eliminated and the problem of
residual solvent in the constructs is solved. The other
advantages of using a water-based binder include the
possibility to incorporate biological agents (e.g. growth
factors) or even living cells [12].
The scaffolds underwent dimensional changes during the
fabrication and post-processing procedures. On the average,
the shrinkage ranged from 2% to 12% depending on the
type of design. The greatest shrinkage was observed from
the circular pore designs.
Treatment at 100 jC for 1 h after printing served to
maintain the integrity and increase the strength of the
scaffolds. Starch is made up predominantly of amylose
and amylopectin, encapsulated in granules. Starch granules
do not dissolve in water, but merely form a suspension. As
granular starch – water suspensions were heated, the starch
granules swell and burst or gelatinise to form a viscous
paste [13]. The amylose partial phase separates from
amylopectin after gelatinisation and forms a continuous
phase that surrounds other swollen granules [14]. As the
starch dispersion cools down, the amylose phase separates,
leading to gel formation and rapid recrystallisation or
‘‘retrogradation’’ [15]. After retrogradation takes place,
the starch has a semicrystalline structure. This double
helical crystalline structure is highly resistant to both
no suspension formed in this study, there was sufficient
moisture and heat for gelatinisation to occur. The work of
Stapley et al. [17] revealed that gelatinisation depended on
two main factors of water and heat. In a starch suspension,
gelatinisation temperature could be about 60 jC. Starch
with its initial water content of about 12% would also
gelatinise, however, at a higher temperature. The end
products also made the removal of unbound powder easier,
as the scaffolds became harder and stiffer after the post-
processing. After heat treatment, the three-blend powdered
particles were sintered together by necking connections
between the particles. Table 1 shows the fabricated scaf-
folds after infiltration.
Percent Dry (g) Second Water Percent
absorbed 10 min (g) absorbed (g) absorbed
97.11 0.283 0.499 0.216 76.59
72.43 0.310 0.497 0.187 60.32
52 C.X.F. Lam et al. / Materials Science and Engineering C 20 (2002) 49–56

Fig. 1. SEM image showing the uniformly coated microstructure for the L3 group copolymer-infiltrated scaffold using 33
magnification.

3.2. Effect of water absorption
Table 2 reveals that, as the volume of the copolymer
used gets larger, the resistance to water absorption becomes
better. After the first 10 min, the infiltrated scaffolds in
group L3 (72.43%) were more resistant than those infil-
trated by L1 and L2. The second 10 min of soaking was
conducted after the scaffolds were soaked in water for 10
min and dried at 100 jC. The results had improved, as it
was observed that the percentage of water absorption of
scaffolds infiltrated by L3 had dropped from 72.43% to
60.32%.
The L3 scaffolds were subjected to a 3-day water-soak-
ing test; this was to evaluate the integrity of the scaffolds for
long hours in a liquid system. The scaffolds lasted the 3
days without much visible surface degradation. Qualita-
tively, the L3 scaffolds were observed to be uniformly
sturdy and strong throughout the test. The SEM image of
the cross-section of an L3 scaffold (Fig. 1) showed an
‘‘undegraded’’ uniform interior. This was attributed to the
uniform infiltration of the L3 specimen.
By using a low initial concentration of 2%, the viscosity
was low enough to enable the copolymer to thoroughly
penetrate the microstructure of the scaffold. As evaporation

Fig. 2. DSC—temp. vs. heat flow for three samples.

 C.X.F. Lam et al. / Materials Science and Engineering C 20 (2002) 49–56 53

Fig. 3. Compressive stiffness of dry scaffolds.

took place, the concentration and viscosity of the copolymer
slowly increased. By diffusion, this copolymer coats the
microstructure of the scaffold both externally and internally.
This infiltration mechanism continuously coated the matrix
with an increasing concentration of the copolymer until
complete evaporation of the solvent.
From the DSC graph in Fig. 2, it can be seen that the
thermal signature of the powder blend had a peak at about
124 jC, while the PLLA – PCL copolymer had a peak at
134 jC. It is known from the literature that the Tm for PLLA
and PCL are about 180 and 60 jC, respectively. According
to Tsuji et al. [18], the peak observed could be ascribed to
the melting of crystallites of LLA-unit sequences in the
copolymer, but not those of CL-unit sequences because of
their different melting temperatures. The peak of the copoly-
mer was observed to be nonsymmetrical and this was
similar to the results of Tsuji et al. [18]. The third thermal
signature was that of the infiltrated scaffold after soaking,
with a peak at 135 jC. This meant that both the peaks from
the powder blend and copolymer had totally disappeared or
merged. Whichever the case, one final peak after the post-
processing indicated that some form of chemical bonding
had taken place. It also showed good mixability between the
materials in forming new bonds. As also shown in Table 2,
the scaffolds were more resistant to water absorption after
the first 10 min of soaking and drying at 100 jC.
3.3. Mechanical properties of scaffolds
Properties, such as stiffness and yield, were evaluated
from the results of compression tests. Fig. 3 showed that the
solid cylinder design had the highest initial stiffness for both
the uninfiltrated and infiltrated scaffolds. While the other
porous designs showed varying degrees of stiffness, a

Fig. 4. Initial yield strength of dry scaffold.

54 C.X.F. Lam et al. / Materials Science and Engineering C 20 (2002) 49–56

Fig. 5. (a) Stiffness of soaked scaffold. (b) Stiffness of soaked scaffold, ‘‘rod_censolid’’.

significant increase in stiffness was observed after initial
infiltration. Also, the ‘‘rod_censolid’’ scaffold of the L3
group resulted in significantly higher stiffness. This dem-
onstrated the effectiveness of the infiltration method in
improving the water resistance and the mechanical proper-
ties of the scaffolds.
The highest initial compressive yield strength for unin-
filtrated and infiltrated scaffolds was also from the solid
cylinder design as can be seen from Fig. 4. However, the
two rather similar yield strengths seemed to suggest that
infiltration did not affect the yield strength for a solid
design. The solid design also displayed cracking and crum-
bling of the external wall, in contrast to the collapse of the
scaffold cross-section as observed from the porous designs.
It was noted that for the porous designs, there was a
significant increase in the yield strength after infiltration.

Table 3
Microporosity of fabricated scaffolds of all designs
Designs Va (m3) Vt (m3), pycnometer Microporosity
6 7
Uninfiltrated Cylinder 1.312
10 5.374
10 0.590
Bar_cenholl 4.88
10  7 2.689
10  7 0.449
Bar_censolid 6.365
10  7 3.646
10  7 0.427
Rod_cenholl 7.064
10  7 3.810
10  7 0.461
Rod_censolid 7.861
10  7 3.929
10  7 0.500
L1 Cylinder 1.312
10  6 5.938
10  7 0.547
Bar_cenholl 4.88
10  7 3.244
10  7 0.335
Bar_censolid 6.365
10  7 4.103
10  7 0.355
Rod_cenholl 7.064
10  7 3.961
10  7 0.439
Rod_censolid 7.861
10  7 4.523
10  7 0.425
L3 Rod_censolid 7.861
10  7 4.507
10  7 0.427
 C.X.F. Lam et al. / Materials Science and Engineering C 20 (2002) 49–56
Fig. 6. Stiffness – porosity relations (dry).
This reinforced the theory that infiltration improved the
mechanical properties of the scaffolds.
The compressive stiffness of soaked (4 h in saline
solution) and infiltrated scaffolds is shown in Fig. 5a and
b. The solid cylinder design showed the highest stiffness
with porous scaffolds having close stiffness readings rang-
ing from 0.059 to 0.102 MPa (Fig. 5a). However, in Fig. 5b,
it was noted that the L3 group has high stiffness after
soaking.
3.4. Porosity analysis
Based on various designs, different porosities were
obtained. It was observed that the fabricated scaffold poros-
ity corresponded to the designed porosities, with some slight
differences due to fabrication side effects such as shrinkage.
Microporosity is unique in powdered materials due to the
voids formed between the granules or particles in the bulk
material. Physically, the voids containing the trapped air can
make way to provide space for the swelling effect when the
scaffolds get wet. Also, it may increase the degradation rate
of the scaffolds due to the increased surface area. While the
comparison between the uninfiltrated and infiltrated scaf-
folds for macroporosity was meaningless, it would be
beneficial to know how the microporosity for the bulk
material was affected after infiltration and also by the
different infiltration methods.
It was observed from Table 3 that the microporosity
decreased after infiltration because the copolymer had
infiltrated the micropores and voids. However, total elimi-
nation of micropores would not be possible as the dichloro-
methane would eventually vaporise and leave some voids
behind.
The graph in Fig. 6 demonstrates the linear and inversely
proportional relationship between the compressive stiffness
and porosity. This mechanical strength and porosity rela-
tionship would be useful in aiding the researcher in deter-
55
mining an optimal scaffold design with the required porosity
and strength.
4. Conclusions
In this study, a new approach to design and fabricate
scaffolds for tissue engineering using the three-dimensional
printing (3DP) in combination with natural polymer and
water-based binder was investigated. Scaffolds were char-
acterised for their physical and chemical properties. The
analysis and tests used in this study demonstrated that
porous 3D scaffolds created by a new blend of materials
through 3DP were achievable. A set of possible fabrication
methodology and post-processing methods for a functional
and interconnected scaffold was also derived. However,
biocompatibility of the set of materials used needs to be
studied in the future. The features and properties of the
present porous scaffolds have shown that they might be
suitable for tissue engineering applications. Different scaf-
fold designs can be applied to produce scaffolds with
repeatable 3D geometry, highly interconnected porous net-
works and suitable mechanical properties.
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