Professional Documents
Culture Documents
W.M.Keck Center for 3D Innovation, The University of Texas at El Paso, El Paso, TX 79968, USA
School of Mechanical Engineering, Pusan National University, San 30 Jangjeon-dong, Geumjeong-gu, Busan 609-735, South Korea
Department of Mechatronics Engineering, Cheju National University, Jejudaehakno, Jeju-Si, Jeju-do 692-756, South Korea
d
Department of Polymer Science and Engineering, Pusan National University, San 30 Jangjeon-dong, Geumjeong-gu, Busan 609-735, South Korea
b
c
a r t i c l e
i n f o
Article history:
Received 19 December 2008
Received in revised form 28 April 2009
Accepted 1 May 2009
Keywords:
Poly(propylene fumarate) (PPF)
Microstereolithography (SL)
Scaffold
a b s t r a c t
Microstereolithography (SL) technology can fabricate three-dimensional (3D) tissue engineered scaffolds with controlled biochemical and mechanical micro-architectures. A SL system for tissue
engineering was developed using a Digital Micromirror Device (DMDTM ) for dynamic pattern generation and an ultraviolet (UV) lamp ltered at 365 nm for crosslinking the photoreactive polymer solution.
The SL system was designed with xy resolution of 2 m and a vertical (z) resolution of 1 m. To
demonstrate the use of SL in tissue engineering, poly(propylene fumarate) (PPF) was synthesized with
a molecular weight of 1200 Da. The viscosity of the PPF was reduced to 150 cP (at 50 C) by mixing
with diethyl fumarate (DEF) in the ratio of 7:3 (w/w). Finally, 2% (w/w) of bis(2,4,6-trimethylbenzoyl)
phenylphosphine oxide (BAPO) was added to the solution to serve as a photoinitiator. Cure depth experiments were performed to determine the curing characteristics of the synthesized PPF, and the resulting
system and prepolymer were used to construct a 3D porous scaffold with interconnected pores of
100 m. Scanning electron microscopy (SEM), and micro-computed tomography (CT) images of the
micro-architecture illustrate that the developed SL system is a promising technology for producing
biodegradable and biocompatible 3D micro-scaffolds with fully interconnected pores.
2009 Elsevier B.V. All rights reserved.
1. Introduction
Microstereolithography (SL) technology, which evolved from
conventional stereolithography, was suggested for producing
micro-scale complex structures. The rst types of SL systems
were based on the vector-based scanning SL method, referred to
as line-scan (Ikuta and Kirowatari, 1993; Takagi and Nakajima,
1993). These laser-based scanning methods also led to the development of nanostereolithography methods using two photon initiated
photopolymerization where several precisely detailed, threedimensional microstructures have been demonstrated (Maruo and
Kawata, 1998; Park et al., 2005; Yang et al., 2007). A number of SL
researchers continue using laser-based scanning systems today,
although much focus of current efforts is in mask-based SL systems, where entire layers are projected at a single time using a
physical mask instead of scanning the surface with a laser beam.
The mask-based SL method was suggested by Nakamoto and
Yamaguchi (1996). Further developments led to a dynamic mask
projection SL system in 1997 by Bertsch et al. where a Liquid Crystal Display (LCD) was used as a dynamic mask instead of the physical
Corresponding author. Tel.: +82 51 510 2327; fax: +82 51 514 0685.
E-mail address: sehlee@pusan.ac.kr (S.-H. Lee).
0924-0136/$ see front matter 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.jmatprotec.2009.05.004
J.-W. Choi et al. / Journal of Materials Processing Technology 209 (2009) 54945503
Table 1
The pore sizes used in the literature.
Literature
Cell/tissue
Osteoblasts
200400
Neovascularization
Fibroblast
Adult mammalian skin
Bone
5
515
20125
100350
Peripheral nerve
Fibrocartilage formation
Liver tissues
Human chondrocytes
60550
100300
45150
190290
scaffold, however, was limited because of the resolution of the system. Lee et al. (2007, 2008) fabricated porous PPF scaffolds with
micro-architecture using a scanning SL system and demonstrated
cell attachment on the fabricated scaffolds. However, the geometries demonstrated by Lee et al. were limited to simple shapes
and true three-dimensionality, particularly with 3D interconnecting channels that are difcult to be fabricated in SL. As a result, the
capability of SL to fabricate complex 3D micro-architectures has
not yet been widely explored, and optimization of these scaffold
geometries requires examination with regard to cell attachment,
proliferation, and differentiation.
The biodegradable and biocompatible scaffold for tissue engineering requires a desired tissue shape, and a porous and
interconnected micro-architecture, which provides a pathway for
transport of nutrients and metabolic waste to and from a cell and/or
tissue. In addition, porous scaffolds should have high surface area
for the initial cell attachment, and a specic micro-scale pore size
depending on cell/tissue types (Hutmacher, 2001; Yang et al., 2001).
Table 1 shows the reported pore sizes according to the cell/tissue
types used in various research. As shown in Table 1, the pore size
depends on the desired cell/tissue type ranging from 5 m to hundreds of microns. This shows the optimal size of the pore requires
investigation using scaffolds with controlled micro-architectures.
These scaffolds can further be used for examination of optimum
pore geometry with respect to cell/tissue type.
In this work, to produce biodegradable and biocompatible scaffolds having a controlled micro-architecture, the development of
a DMDTM (Digital Micromirror Device)-based microstereolithography (SL) system, PPF synthesis and characterization, and
fabrication of 3D PPF micro-scaffolds using medical imaging data
were performed. PPF was synthesized using DEF (diethyl fumarate)
and PG (propylene glycol) along with the hydroquinone and zinc
chloride. The DMD-based SL system was developed with the
lateral resolution of 2 m, and vertical resolution of 1 m.
The synthesized PPF was mixed with the DEF to reduce the PPF
viscosity for use in the SL system. In order to characterize
PPF/DEF, curing tests for penetration depth and critical energy were
performed. Finally, the fabricated microstructures using medical
imaging data were examined by optical microscopy, Scanning electron microscopy (SEM), and micro-computed tomography (CT) to
verify the capability of the developed SL system for producing
micro-architectures with interconnecting micro-pores. The following sections describe these experiments and their results in more
detail.
5495
5496
J.-W. Choi et al. / Journal of Materials Processing Technology 209 (2009) 54945503
Table 2
The original and optimized distances between the optics.
Objective lens
N.A. 0.13
N.A. 0.3
ldt (mm)
lto (mm)
Original distance
Optimized distance
Original distance
Optimized distance
120
120
115.2
115.2
150
150
203.3
186.5
50/115.2 (0.434)
20/115.2 (0.174)
Fig. 3. Building process: (a) irradiation and curing; (b) deep dip traversing so the neighboring solution can ow on top of the part; (c) traversing upward to the desired
position (1 layer thickness); (d) waiting until the solution surface becomes uniform.
J.-W. Choi et al. / Journal of Materials Processing Technology 209 (2009) 54945503
5497
Fig. 4. Fabricated microstructures using the developed SL system: (a) micro-wineglass (Choi, 2007); (b) micro-cup (Choi, 2007); (c) micro-bishops (Ha et al., 2008); (d)
micro-springs (Choi et al., 2009).
Table 3
Fabrication conditions for the microstructures in Fig. 4.
Model
Total layer
number
Layer thickness
(m)
Objective
lens
(a)
(b)
(c)
(d)
300
200
134
300
5
12
20
4
0.3
0.13
0.3
0.3
Material
a
SI40 was purchased from 3D systems (USA), and suitable for conventional SL
system.
b
IBXA represents isobornyl acrylate, and purchased from Woorim Chem Tech Co.,
South Korea.
c
HDDA represents 1,6-hexanediol diacrylate, and purchased from Miwon Commercial Co., South Korea.
d
BED represents bisphenol-A-ethoxylated (4) diacrylate, and purchased from
Hannong Chemicals Co., South Korea.
e
DMPA represents dimethoxy-2-phenylacetophenone, and purchased from
Fisher Scientic Korea, Seoul, South Korea.
5498
J.-W. Choi et al. / Journal of Materials Processing Technology 209 (2009) 54945503
the viscosity of the solution without changing the desired biomaterial properties. DEF, which is a main material for PPF synthesis,
was used as the diluent as it has been reported that DEF does not
signicantly alter the biomaterial properties of PPF. In addition, DEF
has a carbon double bond, so it participates in crosslinking (Fisher
et al., 2002b). In this work, DEF was added to PPF with the ratio of
3:7 (w/w), and the ratio was obtained from the work of Fisher et al.
(2002b). Fisher et al. (2002b) showed that the mixture of DEF/PPF
(3:7 by wt.%), where the molecular weight of the synthesized PPF
was 1260 Da, had the highest elastic modulus and fracture strength.
The viscosity of the PPF/DEF as a function of temperature is shown
in Fig. 6 and was measured using the same viscometer as above. The
PPF/DEF prepolymer was prepared by stirring for 12 h along with
the 2% (w/w) of BAPO as a photoinitiator. In Fig. 6, the viscosity of the
PPF/DEF represents 700 cP at room temperature. This viscosity is
relatively high compared to the desired value of 200 cP. However,
the viscosity of the solution was logarithmically reduced by elevating the temperature, and the viscosity was 150 cP at 50 C. This
value is suitable for SL (Choi, 2007), so a hot plate was installed
under the vat to maintain the temperature of the solution at 50 C.
3.4. Cure depth experiment
A photocurable solution is crosslinked by connecting a
monomer, oligomer or polymer chain, where they have carbon double bonds that can be broken down by a radical. The photons from
the projected light break down the photoinitiator into a radical
along the penetration direction of the light. The radicals then bond
the neighboring monomer, oligomer, or polymer by breaking carbon double bonds. Therefore, in terms of 3D micro-fabrication, the
penetration depth of the light and critical energy at photoinitiation
are important and need to be controlled.
To examine the penetration depth and critical energy, curing
depth experiments were conducted using the PPF/DEF prepolymer.
The energy delivered on the solution surface (Emax ) penetrates into
the solution. The energy inside the solution at the depth z (E(z)) is
dened by BeerLambert law as described in Eq. (1), where Dp is
the penetration depth of the solution (Jacobs, 1993). By introducing
the critical energy (Ec ) into Eq. (1), the curing depth (Cd ) can be
dened as in Eq. (2), where Ec is the energy at the gel point. The
gel point is the point at which solidication begins. Therefore, two
important characteristics of the photocurable solution are Ec and
Dp . These can be experimentally determined through measuring
of the curing depth according to the exposure energy. From the
determined values of Ec and Dp , the exposure energy and stacking
thickness can be chosen (Choi, 2007; Choi et al., 2009). In addition,
it is important to note that the smaller the curing depth, the ability
to fabricate down-facing and complex microstructures is improved:
E(z) = Emax e(z/Dp )
Cd = z(Emax ) = Dp ln
E
max
Ec
(1)
(2)
J.-W. Choi et al. / Journal of Materials Processing Technology 209 (2009) 54945503
5499
Fig. 8. Fabricated crossbeam specimens with exposure times of: (a) 2 s; (b) 3 s; (c) 4 s; (d) 5 s exposure time.
used after the deep dip process, and the SL system was equipped
with the objective lens with the N.A. of 0.3.
Fig. 11 shows the SEM image of the fabricated PPF/DEF kidney
scaffold with a volume of 1400 m 820 m 700 m and pore
size of 100 m. The scaffold was built with 169 layers and the
layer thickness of 4 m. The exterior of the scaffold appears to have
reproduced the 3D geometry quite well, including the freeform
outer shape of the kidney as well as the rectangular vertical pores.
However, the fabricated pore size (100 m) is smaller than the
designed size (120 m) due to material shrinkage. Furthermore,
micro-CT imaging (CT 80, SCANCO Medical AG Co., Switzerland)
was used to show cross-sectional images, internal pore architectures, and interconnectivity. The reconstructed image from the
micro-CT data, shown in Fig. 12, illustrates that the rectangular
pores have good interconnectivity in the vertical direction. However, the horizontal channels could not be found in any of the
micro-CT imaging planes. This could be due to overcure on the
down-facing surfaces, crosslinking of uncured PPF/DEF prepolymer
in the channels due to successive scanning (i.e., similar to overcure,
UV penetration into the uncured trapped material in the channels),
and/or the inability to remove uncured PPF/DEF from the channels
when rinsing and subsequently curing this uncured material in the
post-curing process.
4.3. Discussion
5500
J.-W. Choi et al. / Journal of Materials Processing Technology 209 (2009) 54945503
Fig. 10. Reverse engineering process to reconstruct human kidney model from CT data: (a) CT imaging data; (b) extracted point cloud; (c) cleaned point cloud; (d) crosssectional point series; (e) cross-sectional B-Spline curves; (f) lofted surface; (g) solid geometry model; (h) scaffold model; (i) binary image series used to fabricate the scaffold
in SL.
Fig. 12. Reconstructed micro-CT image of PPF/DEF scaffold: (a) reconstructed image; (b) horizontal cross-sectional image (the scale bar is 100 m).
J.-W. Choi et al. / Journal of Materials Processing Technology 209 (2009) 54945503
Fig. 13. SEM image of fabricated kidney scaffold using the commercial resin.
5501
Fig. 14. Fabricated PPF/DEF cubic scaffold with more pores: (a) SEM image; (b) horizontal cross-sectional CT image.
5502
J.-W. Choi et al. / Journal of Materials Processing Technology 209 (2009) 54945503
Choi, J.W., Ha, Y.M., Won, M.H., Choi, K.H., Lee, S.H., 2005. Fabrication of 3dimensional microstructures using dynamic image projection. In: Proceedings of
International Conference on Precision Engineering and Micro/Nano Technology
in Asia (ASPEN 2005), Shenzhen, China, pp. 472476.
Choi, J.W., Ha, Y.M., Lee, S.H., Choi, K.H., 2006. Design of microstereolithography
system based on dynamic image projection for fabrication of three-dimensional
microstructures. J. Mech. Sci. Technol. 20 (12), 20942104.
Choi, J.W., 2007. Development of projection-based microstereolithography apparatus adapted to large surface and microstructure fabrication for human
body application. Ph.D. Dissertation. Pusan National University, Busan, South
Korea.
Choi, J.W., Wicker, R.B., Cho, S.H., Ha, C.S., Lee, S.H., 2009. Cure depth control for
complex 3D microstructure fabrication in dynamic mask projection microstereolithography. Rapid Prototyping J. 15 (1), 5970.
Cooke, M.N., Fisher, J.P., Dean, D., Rimnac, C., Mikos, A.G., 2002. Use of stereolithography to manufacture critical-sized 3D biodegradable scaffolds for bone ingrowth.
J. Biomed. Mater. Res. B 64B, 6569.
Farsari, M., Claret-Tournier, F., Huang, S., Chatwin, C.R., Budgett, D.M., Birch, P.M.,
Young, R.C.D., Richardson, J.D., 2000. A novel high-accuracy microstereolithography method employing an adaptive electro-optic mask. J. Mater. Process.
Technol. 107, 167172.
Fisher, J.P., Holland, T.A., Dean, D., Engel, P.S., Mikos, A.G., 2001. Synthesis and properties of photocross-linked poly(propylene fumarate) scaffolds. J. Biomater. Sci.
Polym. Ed. 12 (6), 673687.
Fisher, J.P., Vehof, J.W.M., Dean, D., Waerden, J.P., Holland, T.A., Mikos, A.G., Jansen,
J.A., 2002a. Soft and hard tissue response to photocrosslinked poly(propylene
fumarate) scaffolds in a rabbit model. J. Biomed. Mater. Res. 59, 547
556.
Fisher, J.P., Dean, D., Mikos, A.G., 2002b. Photocrosslinking characteristics and
mechanical properties of diethyl fumarate/poly(propylene fumarate) biomaterials. Biomaterials 23, 43334343.
Ha, C.S., Jung, S.J., Kim, E.S., Kim, W.S., Lee, S.J., Lee, S.J., 1996. Properties of UV-curable
polyurethane acrylates using nonyellowing polyisocyanate for oor coating. J.
Appl. Polym. Sci. 62, 10111021.
Ha, Y.M., Choi, J.W., Lee, S.H., 2008. Mass production of 3-D microstructures using
projection microstereolithography. J. Mech. Sci. Technol. 22 (3), 514521.
Hadlock, T., Sundback, C., Hunter, D., Cheney, M., Vacanti, J.P., 2000. A polymer foam
conduit seeded with Schwann cells promotes guided peripheral nerve regeneration. Tissue Eng. 6 (2), 119127.
Han, L.H., Mapili, G., Chen, S., Roy, K., 2008. Projection microfabrication of threedimensional scaffolds for tissue engineering. J. Manuf. Sci. Eng. Trans. ASME 130
(2), 021005-1-4.
Hutmacher, D.W., 2001. Scaffold design and fabrication technologies for engineering
tissuesstate of the art and future perspectives. J. Biomater. Sci. Polym. Ed. 12
(1), 107124.
Ikuta, K., Kirowatari, K., 1993. Real three dimensional micro fabrication using
stereo lithography and metal molding. In: Proceedings of 6th IEEE Workshop
on Micro Electro Mechanical Systems (MEMS93), New York, NY, USA, pp.
4247.
Jacobs, P.F., 1993. Rapid prototyping and manufacturing: fundamentals of stereolithography. Soc. Manuf. Eng..
Jung, S.J., Lee, S.J., Cho, W.J., Ha, C.S., 1998. Synthesis and properties of UV-curable
waterborne unsaturated polyester for wood coating. J. Appl. Polym. Sci. 69,
695708.
Lee, J.W., Lan, P.X., Kim, B., Lim, G., Cho, D.W., 2007. 3D scaffold fabrication with
PPFDEF using micro-stereolithography. Microelectron. Eng. 84, 17021705.
Lee, J.W., Lan, P.X., Kim, B., Lim, G., Cho, D.W., 2008. Fabrication and characteristic
analysis of a poly(propylene fumarate) scaffold using micro-stereolithography
technology. J. Biomed. Mater. Res. B 87B (1), 19.
Leong, K.F., Cheah, C.M., Chua, C.K., 2003. Solid freeform fabrication of threedimensional scaffolds for engineering replacement tissues and organs.
Biomaterials 24, 23632378.
Limaye, A.S., Rosen, D.W., 2007. Process planning method for mask projection microstereolithography. Rapid Prototyping J. 13 (2), 7684.
Maruo, S., Kawata, S., 1998. Two-photon-absorbed near-infrared photopolymerization for three-dimensional microfabrication. J. Microelectromech. Syst. 7 (4),
411415.
Menon, A.R.R., Aigbodion, A.I., Pillai, C.K.S., Mathew, N.M., Bhagawan, S.S., 2002. Processability characteristics and physicmechanical properties of natural rubber
modied with cashewnut shell liquid and cashewnut shell liquid-formaldehyde
resin. Eur. Polym. J. 38, 163168.
Nakamoto, T., Yamaguchi, K., 1996. Consideration on the producing of high
aspect ratio micro parts using UV sensitive photopolymer. In: Proceedings
of 7th International Symposium on Micro Machine and Human Science, pp.
5358.
Park, S.H., Lee, S.H., Yang, D.Y., 2005. Subregional slicing method to increase threedimensional nanofabrication efciency in two-photon polymerization. Appl.
Phys. Lett. 87, 154108.
Peter, S.J., Kim, P., Yasko, A.W., Yaszemski, M.J., Mikos, A.G., 1999. Crosslinking characteristics of an injectable poly(propylene fumarate)/-tricalcium phosphate paste
and mechanical properties of the crosslinked composite for use as a biodegradable bone cement. J. Biomed. Mater. Res. 44, 314321.
Shung, A.K., Timmer, M.D., Jo, S., Engel, P.S., Mikos, A.G., 2002. Kinetics of
poly(propylene fumarate) synthesis by step polymerization of diethyl fumarate
and propylene glycol using zinc chloride as a catalyst. J. Biomater. Sci. Polym. Ed.
13 (1), 95108.
J.-W. Choi et al. / Journal of Materials Processing Technology 209 (2009) 54945503
Sun, C., Fang, N., Wu, D.M., Zhang, X., 2005. Projection micro-stereolithography
using digital micro-mirror dynamic mask. Sens. Actuator A: Phys. 121 (1),
113120.
Takagi, T., Nakajima, N., 1993. Photoforming applied to ne machining. In: Proceedings of 4th International Symposium on Micro Machine and Human Science
(MHS93), pp. 173178.
Tienen, T.G., Heijkants, R.G.J.C., Buma, P., Groot, J.H., Pennings, A.J., Veth, R.P.H., 2002.
Tissue ingrowth and degradation of two biodegradable porous polymers with
different porosities and pore sizes. Biomaterials 23, 17311738.
Varadan, V.K., Jiang, X., Varadan, V.V., 2001. Microstereolithography and Other Fabrication Techniques for 3D MEMS. John Wiley & Sons Ltd.
5503
Whang, K., Healy, K.E., Elenz, D.R., Nam, E.K., Tsai, D.C., Thomas, C.H., Nuber, G.W., Glorieux, F.H., Travers, R., Sprague, S.M., 1999. Engineering bone regeneration with
bioabsorable scaffolds with novel microarchitecture. Tissue Eng. 5 (1), 3551.
Wu, D.M., 2005. Micro fabrication of 3D structures and characterization of molecular
machine. Ph.D. Dissertation. UCLA, Los Angeles.
Yang, D.Y., Park, S.H., Lim, T.W., Kong, H.J., Yi, S.W., Yang, H.K., Lee, K.S., 2007. Ultraprecise microreproduction of a three-dimensional artistic sculpture by multipath
scanning method in two-photon photopolymerization. Appl. Phys. Lett. 90,
079903.
Yang, S., Leong, K.F., Du, Z., Chua, C.K., 2001. The design of scaffolds for use in tissue
engineering. Part I. Traditional factors. Tissue Eng. 7 (6), 679689.