The Biomaterials Silver Jubilee C o m p e n d i u m 175

Biomaterials
ELSEVIER Biomaterials 21 (2000) 2529-2543

Scaffolds in tissue engineering bone and cartilage

Dietmar W. Hutmacher
Laboratory for Biomedical Engineering, Institute of Engineering Science, Department of Orthopedic Surgery,
National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260, Singapore

Abstract

Musculoskeletal tissue, bone and cartilage are under extensive investigation in tissue engineering research. A number of
biodegradable and bioresorbable materials, as well as scaffold designs, have been experimentally and/or clinically studied. Ideally,
a scaffold should have the following characteristics: (i) three-dimensional and highly porous with an interconnected pore network for
cell growth and flow transport of nutrients and metabolic waste; (ii) biocompatible and bioresorbable with a controllable degradation
and resorption rate to match cell/tissue growth in vitro and/or in vivo; (iii) suitable surface chemistry for cell attachment, proliferation,
and differentation and (iv) mechanical properties to match those of the tissues at the site of implantation. This paper reviews research
on the tissue engineering of bone and cartilage from the polymeric scaffold point of view. 9 2000 Elsevier Science Ltd. All rights
reserved.

Keywords" Tissue engineering of bone and cartilage; Design and fabrication of 3-D scaffold; Biodegradable and bioresorbable polymers

1. Introduction The currently applied scaffold fabrication technologies,

Bone and cartilage generation by autogenous cell/tis-
sue transplantation is one of the most promising tech-
niques in orthopedic surgery and biomedical engineering
[1]. Treatment concepts based on those techniques
would eliminate problems of donor site scarcity, immune
rejection and pathogen transfer [-2]. Osteoblasts, chon-
drocytes and mesenchymal stem cells obtained from the
patient's hard and soft tissues can be expanded in culture
and seeded onto a scaffold that will slowly degrade and
resorb as the tissue structures grow in vitro and/or
in vivo [3]. The scaffold or three-dimensional (3-D) con-
struct provides the necessary support for cells to prolifer-
ate and maintain their differentiated function, and its
architecture defines the ultimate shape of the new bone
and cartilage. Several scaffold materials have been inves-
tigated for tissue engineering bone and cartilage includ-
ing hydroxyapatite (HA), poly(~-hydroxyesters), and
natural polymers such as collagen and chitin. Several
reviews have been published on the general properties
and design features of biodegradable and bioresorbable
polymers and scaffolds [4-12]. The aim of this paper is to
complete the information collected so far, with special
emphasis on the evaluation of the material and design
characteristics which are of specific interest in tissue
engineering the mesenchymal tissues bone and cartilage.
with special emphasis on the so-called solid-free form
fabrication technologies, will also be bench marked. Fi-
nally, the paper discusses the author's research on the
design and fabrication of 3-D scaffolds for tissue engi-
neering an osteochondral transplant.
2. Polymer-based scaffold materials
The meaning and definition of the words biodegrad-
able, bioerodable, bioresorbable and bioabsorbable
(Table 1)--which are often used misleadingly in the tissue
engineering literature--are of importance to discuss the
rationale, function as well as chemical and physical proper-
ties of polymer-based scaffolds. In this paper, the polymer
properties are based on the definitions given by Vert [13].
The tissue engineering program for bone and cartilage
in the author's multidisciplinary research curriculum has
been classified into six phases (Table 2). Each tissue
engineering phase must be understood in an integrated
manner across the research program--from the polymer
material properties, to the scaffold micro- and macro-
architecture, to the cell, to the tissue-engineered trans-
plant, to the host tissue. Hence, the research objectives in
each phase are cross-disciplinary and the sub-projects are
linked horizontally as well as vertically.

0142-9612/00/$- see front matter 9 2000 Elsevier Science Ltd. All rights reserved.
PII: S0 142-96 12(00)001 2 1-6
 176
2530 D. W. Hutmacher / Biomaterials 21 (2000) 2529-2543
Table 1
Definitions given by Vert
Biodegradable are solid polymeric materials and devices which break
down due to macromolecular degradation with dispersion in vivo but
no proof for the elimination from the body (this definition excludes
environmental, fungi or bacterial degradation). Biodegradable poly-
meric systems or devices can be attacked by biological elements so that
the integrity of the system, and in some cases but not necessarily, of the
macromolecules themselves, is affected and gives fragments or other
degradation by-products. Such fragments can move away from their
site of action but not necessarily from the body.
Bioresorbable are solid polymeric materials and devices which show
bulk degradation and further resorb in vivo; i.e. polymers which are
eliminated through natural pathways either because of simple filtration
of degradation by-products or after their metabolization. Bioresorption
is thus a concept which reflects total elimination of the initial foreign
material and of bulk degradation by-products (low molecular weight
compounds) with no residual side effects. The use of the word 'bio-
resorbable' assumes that elimination is shown conclusively.
Bioerodible are solid polymeric materials or devices, which show sur-
face degradation and further, resorb in vivo. Bioerosion is thus a con-
cept, too, which reflects total elimination of the initial foreign material
and of surface degradation by-products (low molecular weight com-
pounds) with no residual side effects.
Bioabsorbable are solid polymeric materials or devices, which can
dissolve in body fluids without any polymer chain cleavage or molecu-
lar mass decrease. For example, it is the case of slow dissolution of
water-soluble implants in body fluids. A bioabsorbable polymer can be
bioresorbable if the dispersed macromolecules are excreted.
Table 2
The research program for tissue engineering bone and cartilage classi-
fied into six phases
I--Fabrication of bioresorbable scaffold
II--Seeding of the osteoblasts/chondrocytes populations into the
polymeric scaffold in a static culture (petri dish)
III--Growth of premature tissue in a dynamic environment
(spinner flask)
IV--Growth of mature tissue in a physiologic environment (bioreactor)
V--Surgical transplantation
VI--Tissue-engineered transplant assimilation/remodeling
The first stage of tissue engineering bone or cartilage
begins with the design and fabrication of a porous 3-D
scaffold, the main topic of this review paper. In general,
the scaffold should be fabricated from a highly biocom-
patible material which does not have the potential to
elicit an immunological or clinically detectable primary
or secondary foreign body reaction [9]. Furthermore,
a polymer scaffold material has to be chosen that will
degrade and resorb at a controlled rate at the same time
as the specific tissue cells seeded into the 3-D construct
attach, spread and increase in quantity (number of
cells/per void volume) as well as in quality. Currently, the
design and fabrication of scaffolds in tissue engineering
research is driven by three material categories: I. Regula-
tory approved biodegradable and bioresorbable poly-
mers (Table 3), such as collagen, polyglycolide (PGA),
The Biomaterials Silver Jubilee Compendium
polylactides (PLLA, PDLA), polycaprolactone (PCL),
etc. II. A number of non-approved polymers, such as
polyorthoester (POE), polyanhydrides, etc. which are
also under investigation. III. The synthesis of entrepre-
neurial polymeric biomaterials, such as poly (lactic acid-
co-lysine), etc., which can selectively shepherd specific cell
phenotypes and guide the differentiation and prolifer-
ation into the targeted functional premature and/or ma-
ture tissue.
In general, polymers of the poly(~-hydroxy acids)
group undergo bulk degradation. The molecular weight
of the polymer commences to decrease on day one (PGA,
PDLA) or after a few weeks (PLLA) upon placement in
an aqueous media [12]. However, the mass loss does not
start until the molecular chains are reduced to a size
which allows them to freely diffuse out of the polymer
matrix [ 14]. This phenomenon described and analyzed in
detail by a number of research groups [15-18], results in
accelerated degradation and resorption kinetics until the
physical integrity of polymer matrix is compromised. The
mass loss is accompanied by a release gradient of acidic
by-products.
In vivo, massive release of acidic degradation and
resorption by-products results in inflammatory reac-
tions, as reported in the bioresorbable device literature
[19-22]. If the capacity of the surrounding tissue to
eliminate the by-products is low, due to the poor vas-
cularization or low metabolic activity, the chemical com-
position of the by-products may lead to local temporary
disturbances. One example of this is the increase of osmo-
tic pressure or pH manifested through local fluid
accumulation or transient sinus formation from fiber
reinforced polyglycolide pins applied in orthopedic sur-
gery [21]. Potential problems of biocompatibility in
tissue engineering bone and cartilage, by applying degra-
dable, erodable, and resorbable polymer scaffolds, may
also be related to biodegradability and bioresorbability.
Therefore, it is important that the 3-D scaffold/cell con-
struct is exposed at all times to sufficient quantities of
neutral culture media, especially during the period where
the mass loss of the polymer matrix occurs.
The incorporation of a tricalciumphosphate (TCP)
[23], hydroxyapatite (HA) [24] and basic salts [15] into
a polymer matrix produces a hybrid/composite material.
These inorganic fillers allow to tailor the desired degra-
dation and resorption kinetics of the polymer matrix.
A composite material would also improve biocompatibil-
ity and hard tissue integration in a way that ceramic
particles, which are embedded into the polymer matrix,
allow for increased initial flash spread of serum proteins
compared to the more hydrophobic polymer surface [9].
In addition, the basic resorption products of HA or TCP
would buffer the acidic resorption by-products of the
aliphatic polyester and may thereby help to avoid the
formation of an unfavorable environment for the cells
due to a decreased pH [15,23,24].
 t~

Table 3 t,,,,~ o
Properties of bioresorbable and bioerodable polymers

Polymer Comparison of Degradation and Molecular weight Mass loss References References Area of Products with
mechanical properties resorption process loss/loss of (in month) a (scaffolds) (medical device) application regulatory
of bioerodable and via hydrolysis mechanical properties approval
bioresorbable polymers (in month) a
,~,~o

Poly(L-lactide) + + + Bulk erosion 9-15 36-48 46, 48, 49, 19, 20, 24 Orthopedic FixSorb System
60-64, 70-72 Surgery, Oral and (screws, nails, pins)
Maxillofacial Neofix (screws,
Surgery nails, pins)
Poly(L-lactide-co-D, + + Bulk erosion 5-6 12-18 22, 23 Oral and ResorPin, Leadfix
L-lactide) 70/30 Maxillofacial MacroSorb System
Surgery, Orthopedic (screws and plates,
Surgery mesh, nails, pins)
PolyPin
Poly(L-lactide-co- + + Bulk erosion 1-2 3-4 28, 63 Suture Periodontal Vicryl Suture,
glycolide) 10/90 Surgery, Surgery, Vicryl Mesh
Polyglycolide + + + Bulk erosion 0.5-1 3-4 6, 30-35, 41, 65 Orthopedic Surgery Biofix
Poly(D,L-lactide) + Bulk erosion 1-2 5-6 6, 31, 34, 49, 60 t..j

Poly(D,L-lactide- + Bulk erosion 1-2 4-5 53-56, 60, 70

co-glycolide) 85/15
Poly(D,L-lactide- + Bulk erosion 1-2 4-5 49, 61
co-glycolide) 75/25 t~
t-.a
Poly(D,L-lactide- + + Bulk erosion 1-2 3-4 28 I
co-glycolide) 50/50
4~
Polycaprolactone + Bulk and surface 9-12 24-36 29 Drug delivery Capranor
erosion
Polyorthoester + + Surface erosion 4-6 12-18
Polyanhydrides + + Surface erosion 4-6 12-18 59 Animal experiments

a Molecular weight and mass loss vary depending on factors such as chemical structure and composition; presence of ionic groups and of side group defects; configuration of the structure molecular

weight and molecular weight distribution (polydispersity); presence of low molecular weight components (monomers, oligomers, solvents, softeners, drugs, growth factors, etc.); production and
manufacturing procedures and their process parameters, implant design, sterilization method, morphology (amorphous versus semi-crystalline, presence of microstructures and stress within the
components), tempering, storage, implant site. + + + , good; + + , average; +, poor.

-.q

t.J "q
t.a.a
 178 The Biomaterials Silver Jubilee Compendium
2532 D. W. Hutmacher / Biomaterials 21 (2000) 2529-2543

Control of the hydrodynamic and biochemical envi- the scaffold matrix must serve an additional function; it
ronment is essential for the successful in vitro engineering must provide sufficient temporary mechanical support to
of 3-D scaffold/tissue constructs for potential clinical use withstand in vivo stresses and loading. In Strategy I re-
[25]. Computer-controlled bioreactors that continuously search programs, the material must be selected and/or
supply physiological nutrients and gases, serve to regu- designed with a degradation and resorption rate such
late the required cell/tissue culture conditions for a long that the strength of the scaffold is retained until the tissue
period of time. After the in vitro culturing of the 3-D engineered transplant is fully remodeled by the host
scaffold/tissue construct, the degree of remodeling and tissue and can assume its structural role.
cell/tissue replacement of the bone/cartilage transplant Bone is able to remodel in vivo under so-called physio-
by the host tissue has to been taken into consideration logical loading [27]. It is a requirement that the degrada-
[26]. Cell and tissue remodeling is important for achiev- tion and resorption kinetics have to be controlled in such
ing stable mechanical conditions and vascularization at a way that the bioresorbable scaffold retains its physical
the host site. Hence, the 3-D scaffold/tissue construct properties for at least 6 months (4 months for cell cultur-
should maintain sufficient structural integrity during the ing and 2 months in situ). Thereafter, it will start losing its
in vitro and/or in vivo growth and remodeling process. mechanical properties and should be metabolized by the
The degree of remodeling depends on the host anatomy body without a foreign body reaction after 12-18 months
and physiology [26]. The polymer selection from a ma- (Fig. 1). The mechanical properties of the bioresorbable
terial science point of view is based on two different 3-D scaffold/tissue construct at the time of implantation
strategies in regard to the overall function of the scaffold. should match that of the host tissue as closely as possible
[7]. It should posses sufficient strength and stiffness to
2.1. Strategy I function for a period until in vivo tissue ingrowth has
replaced the slowly vanishing scaffold matrix.
In the first strategy (Fig. 1), the physical scaffold struc- Thompson et al. [28] studied a poly(D,L-lactide-co-
ture supports the polymer/cell/tissue construct from the glycolide) matrix under cyclic compressive loading.
time of cell seeding up to the point where the hard tissue They concluded that changes in surface deformation and
transplant is remodeled by the host tissue. In the case of morphology suggest that the compressive loading ini-
load-bearing tissue such as articular cartilage and bone, tially collapses and stiffens the polymer matrix. The de-

I Hydration
II Hydration and Degradation
III Degradation and Mass Loss
IV Resorption and Metabolisation
I I I V Metabolisation
I II 1 III I IV 1 V
100% i
i
I i
i
I
!
l i
,' Mdle~zular w~ight ios~ ,
i , I i
, ' ~D Scat~fold t
=.-.K,.r-.~,
, \ ,
~, .
, \/
. , . ~ .
,
- . ~ . - , . - , olOSS Ol
lvlass ~ ,, - .
' i\
. . . .
i [ x~ I
i i '1 i

b,oresor~able
,
I
/\.
'

3I~,, scaffol
~ ,

Remodelin", g of tissue
'
.. . .. . . . \]......I- ,,
", ,' / henn g~
~ mt ee;s:ed
e r e d ~trans
; ; ; P ~P lant b
;intn bo
50% l , I i ~ 11 I i / / i _

', I I I "~[', ] ~ k e l e t ~ l tissue in vi

i , ' , ~ I / /

, i ' i ,'.\ I f,
',I !, ',i i! I X".\
I ~ ~I I / 'i
| ! ! I i i

i i 9 i ~ i

0%
! i i I !

i i I i

i0 1 2 ~ 6 9 12 15 24
, ,,
,' ', Numb, er of weeks
I ! !
i
, B C D ', F 'o
I I I
iT M f-~, ~ -,q

A E

Fig. 1. Graphical illustration of the complex interdependence of molecular weight loss and mass loss of a strategy 1 3D scaffold matrix plotted against
the time frame for tissue engineering a cartilage/bone transplant. (A) Fabrication of bioresorbable scaffold; (B) seeding of the osteoblast/cartilage
populations into the polymeric scaffold in a static culture (petri dish); (C) growth of premature tissue in a dynamic environment (spinner flask);
(D) growth of mature tissue in a physiologic environment (bioreactor); (E) surgical transplantation; (F) tissue-engineered transplant assimila-
tion/remodeling.
 The Biomaterials Silver Jubilee Compendium 179
D.W. Hutmacher / Biomaterials 21 (2000) 2529-2543 2533

crease in molecular weight is slowed down due to the
reduction of surface area from hydrolysis, until the
matrix architecture no longer accommodates the mech-
anical loading and begins to lose its integrity. Conclus-
ively, in Strategy I the scaffold architecture has to
withstand mechanical loading in vitro and in vivo.
2.2. Strategy H
For Strategy II (Fig. 2), the intrinsic mechanical prop-
erties of the scaffold architecture templates the cell prolif-
eration and differentiation only up to the phases where
the premature bone or cartilage is placed in a bio-
reactor. The degradation and resorption kinetics of the
scaffold are designed to allow the seeded cells to prolifer-
ate and secrete their own extracellular matrix in the static
and dynamic cell seeding phase (weeks 1-12), while the
polymer scaffold gradually vanishes leaving sufficient
space for new cell and tissue growth. The physical sup-
port by the 3-D scaffold is maintained until the engineer-
ed tissue has sufficient mechanical integrity to support
itself.
Different research groups [29-33] have shown in
a number of studies that a nonwoven mesh made of
polyglycolide fibers offers degradation and resorption
kinetics for Strategy II. However, the challenge for the
grown cell/tissue construct is to have similar mechanical
properties to the host bone and cartilage. Ma and Langer
[34] showed that cartilage which was cultured for
7 month in a bioreactor reached 40% of the mechanical
properties of natural cartilage. The next phase of those
research programs will be to evaluate how these tissue-
engineered cartilage transplants assimilate and remodel
in vivo [35]. In an in vivo model, one of the major
problems from a biomechanical and clinical view point, is
the primary mechanical stabilization of cartilage trans-
plants [28]. This aspect will be discussed below, under
scaffold design, in more detail.
3. Scaffold design
Skeletal tissue, such as bone and cartilage, is usually
organized into 3-D structures in the body [36]. For the
repair and generation of hard and ductile tissue, such as
bone, scaffolds need to have a high elastic modulus in
order to be retained in the space they were designated for;
and also provide the tissue with adequate space for
growth [37]. If the 3-D scaffold is used as a temporary
load-bearing device (Strategy II), the mechanical proper-
ties would maintain that load for the required time with-
out showing symptoms of fatigue or failure. Therefore,
one of the basic problems from a scaffold design point of
view, is that to achieve significant strength the scaffold
material must have sufficiently high interatomic and in-
termolecular bonding, but must have at the same time
a physical and chemical structure which allows for hy-
drolytic attack and breakdown.

[ Hydration Hydration .] Bulk Erosion .] Bulk Erosion .] Metabolisation

[ i Degradation [ Mass loss i Metabolisation
.
9 | .

I I ',
i
I I I
9 . i 9 9

00% i_ . . . . . . ,:

: Molelzula \ 9construct r e n i o d e l i n g a n d
I w;igl~t los; ! /ii ~ //;\ ~ell/tissue repla[~ement by the
: ] ' i / I i ~ / 9i host tissue
I9 ,, !9/ 1 ! ~ \',: ":

50~176i i / Mass l~ ~f'~3 D / s c a f f ~ I

9 , 9 construct in:vitro : :
I)% :i
I "i I ', '1 I Ii
I~ i
v , 3 , 6 12 ',15
i !
'
I
,
I
Number of mo~hs
IB C', D ', F ', i

! w"i'~

'E

Fig. 2. Graphical illustration of the complex interdependence of molecular weight loss and mass loss of a strategy II 3D scaffold matrix plotted against
the time frame for tissue engineering a cartilage/bone transplant. (A) Fabrication of bioresorbable scaffold; (B) seeding of the osteoblast/cartilage
populations into the polymeric scaffold in a static culture (petri dish); (C) growth of premature tissue in a dynamic environment (spinner flask);
(D) growth of mature tissue in a physiologic environment (bioreactor); (E) surgical transplantation; (F) tissue-engineered transplant assimila-
tion/remodeling.
 180
2534 D. W. Hutmacher / Biomaterials 21 (2000) 2529-2543
Ingber and his group [38,39] design their scaffolds
based on a concept which they named tensegrity.
Geodesical 3-D constructs were designed by applying
tensegrity so that the entire scaffold structure evenly
distributes and balances mechanical stresses. The walls,
layers or struts that make up the interconnecting scaffold
framework are connected into triangles, pentagons or
hexagons, each of which can bear tension or compres-
sion. However, the mechanical rational of the tensegrity
design concept has been known for centuries in the area
of civil engineering. Gothic architects used a stone skel-
eton structure of stone columns and ribs supported by
arches and buttresses to build cathedrals (Fig. 3).
Another point, which has to be focused on is the
diffusion of nutrients into the 3-D scaffold. Although, an
interconnected macropore-structure of 300-500 ~tm en-
hances the diffusion rates to and from the center of
a scaffold, transportation of the nutrients and by-prod-
ucts is not sufficient for large scaffold volumes. A fluid-
dynamic microenvironment provided by a bioreactor can
mimic the interstitial fluid conditions present in natural
bone and cartilage in a macroporous scaffold architec-
ture. Bioreactors permit in vitro culture of larger and
better organized 3-D cell communities than can be
achieved using standard tissue culture techniques [40].
Fig. 3. The Cologne cathedral was built in the 18th and 19th century by
gothic architects who used a stone skeleton structure of stone columns
and ribs supported by arches and buttresses.
The Biomaterials Silver Jubilee Compendium
For tissue engineering a bone transplant, the creation
of a vascularized bed ensures the survival and function of
seeded cells, which have access to the vascular system for
nutrition, gas exchange, and elimination of by-products
[41]. The vascularization of a scaffold may be compro-
mised by purely relying on capillary ingrowth into the
interconnecting pore network from the host tissue.
In situ, the distance between blood vessels and mesen-
chymal cells are not larger then 100 gm [42]. Therefore,
the time frame has to be taken into account for the
capillary system to distribute through larger scaffold
volume. It may also be possible to control the degree and
rate of vascularization by incorporating angiogenic and
anti-angiogenic factors in the degrading matrix of the
scaffold.
From a biomechanical and clinical point of view, the
tissue-engineered bone or cartilage transplant should
allow for a mechanically secure and stable fixation on or
to the host tissue [29]. For bone, the currently available
medical devices, such as pins, screws, and plates might be
used. However, the integration of a device-like part into
the 3-D scaffold design can be advantageous. The ration-
ale for such an innovative design concept will be for the
tissue engineering of an osteochondral bone transplant is
described below.
4. Scaffold fabrication
A number of fabrication technologies have been
applied to process biodegradable and bioresorbable
materials into 3-D polymeric scaffolds of high porosity
and surface area. The conventional techniques for scaf-
fold fabrication include fiber bonding, solvent casting,
particulate leaching, membrane lamination and melt
molding (Table 4). Several papers have reviewed the past
and current research on scaffold fabrication techniques
[43-45]. However, none of those papers has directly
compared the 3-D scaffold-processing technologies for
the tissue engineering community. From a scaffold design
and function view point each processing methodology
has its pro and cons. It is the aim of this paper to
aggregate the compiled information and to present this
data in a comprehensive form. Table 4 summarizes the
key characteristics and parameters of the techniques cur-
rently used. The aim of this part of the review paper is to
assist research teams with their choice for a specific 3-D
scaffold-processing technology by providing the informa-
tion needed to determin the critical parameters. As dis-
cussed above, at present the challenge in tissue
engineering bone and cartilage is not only to design, but
also to fabricate reproducible bioresorbable 3-D scaf-
folds, which are able to function for a certain period of
time under load-bearing conditions.
Solvent casting, in combination with particle leaching,
works only for thin membranes or 3-D specimens with
 e~
t,,~~

Table 4
Currently applied 3D scaffold fabrication technologies
c,z
Fabrication technology Processing Material properties Scaffold design and Achievable pore Porosity Architecture Reference
required for reproducibility size in Jam in %
processing
t...~ o
Solvent casting in Casting Soluble User, material and 30-300 20-50 Spherical pores, salt particles remain 47
t~
combination with technique sensitive in matrix
particular leaching
Membrane lamination Solvent bonding Soluble User, material and 30-300 <85 Irregular pore structure 48
technique sensitive
Fabrication of non-woven Carding, Needling, Fibres Machine controlled 20-100 <95 Insufficient mechanical properties 30-35, 41,
Plate pressing 63-65
Melt moulding Moulding Thermoplastic Machine controlled 50-500 < 80 r~

Extrusion in combination Extrusion Thermoplastic Machine controlled < 100 < 84 Spherical pores, salt particles remain 49
with particular leaching through dies in matrix
Emulsion freeze drying Casting Soluble User, material and < 200 <97 High volume of inter-connected 54-56
technique sensitive micropore structure
Thermally induced Casting Soluble User, material and < 200 <97 High volume of inter-connected 58-61
phase separation technique sensitive micropore structure t..a

Supercritical-fluid Casting Amorphous Material and < 100 10-30 High volume of non interconnected 53
technology technique sensitive micropore structure
Supercritical-fluid Casting Amorphous Material and Micropores < 50 <97 Low volume of non-interconnected 54
t-.a
technology in technique sensitive macropores < 400 micropore structure combined t~
t-,a
combination with with interconnected
t..a
particle leaching macropore structure t~
4~
3-D printing in and without Solid free form Soluble Machine and 45-150 < 60 100% interconnected macropore 69-72 t~

combination of particle
leaching
Fused deposition modelling
fabrication computer controlled (triangles, pentagons, honey comb, etc.),
design and fabrication layer by layer,
by use of water-based binder incorpo-
ration of biological agents into
matrix possible
Solid free form Thermoplastic Machine and > 150 < 80 100% interconnected macropore structure 29
fabrication computer controlled (triangles, pentagons, honey comb, etc.),
design and fabrication layer by layer

c~
t,3
 182 The Biomaterials Silver Jubilee C o m p e n d i u m
2536 D.W. Hutmacher/ Biomaterials 21 (2000) 2529-2543

very thin wall sections: otherwise, it is not possible to fabrication of a truly interconnecting pore structure de-
remove the soluble particles from within the polymer pends on the processing method and parameters as well
matrix [46]. Mikos et al. [47], using the above-described as on the used equipment [57,58].
technology, fabricated porous sheets and laminated them Several groups [57-60] studied thermally induced
to 3-D structures. Chloroform was used on the attach- phase separation technology to process polymeric 3-D
ment interface for the lamination process. This fabrica- scaffolds. This technique has been used previously to
tion technology is time consuming because only thin fabricate synthetic membranes for non-medical applica-
membranes can be used. Another disadvantage is that the tions. The method has been extensively applied in the
layering of porous sheets allows only a limited number of field of drug delivery to fabricate microspheres, which
interconnected pore networks. Solvent-casted polymer- allows the incorporation of pharmaceutical and biolo-
salt composites have also been extruded into a tubular gical agents, such as bone morphogenetic proteins
geometry [48]. The disadvantages of the above technolo- (BMPs) into the polymer matrix. In general, the micro-
gies include the extensive use of highly toxic solvents, and macro-structure is controlled by varying the polymer
time required for solvent evaporation (days-to-weeks), material, polymer concentration, quenching temper-
the labor intensive fabrication process, the limitation to ature, and solvents. However, current research shows
thin structures, residual particles in the polymer matrix, that the method, similar then emulsion freeze-drying
irregularly shaped pores, and insufficient interconnectivity. technique, is user and technique sensitive and that the
The supercritical fluid-gassing process has been known processing parameters have to be well controlled. Nam
for many years in the non-medical polymer industry [49] and Park [57] as well as Zhang and Ma [58] fabricated
as well as in the pharmaceutical community [50]. This polymer and polymer/HA specimens with a porosity of
technology is used to produce foams and other highly
porous products. The polymers which can be used for
this technology have to have an high amorphous frac-
tion. The polymer granules are plasticized due to the
employment of a gas, such as nitrogen or carbon dioxide,
at high pressures. The diffusion and dissolution of the gas
into the polymer matrix results in a reduction of the
viscosity, which allows the processing of the amorphous
bioresorbable polyesters in a temperature range of
30-40~ [51]. The supercritical fluid-gassing technology
allows the incorporation of heat sensitive pharmaceut-
icals and biological agents. However, on average only
10-30% of the pores are interconnected [51,52]. Harris
et al. [53] combined this technology with particulate
leaching to gain a highly interconnected void network.
The researchers could control porosity and pore size by
varying the particle/polymer ratio and particle size.
Whang et al. [54,55] developed a protocol for the
fabrication of aliphatic polyester-based scaffolds by using Fig. 4. Polymerdisks with a diameter of 500 ~tm and 40 gm thickness
the emulsion freeze-drying method. Scaffolds with poros- allow to stack a 3D scaffold with a porosity of 98%.
ity greater than 90%, median pore sizes ranging from 15
to 35 gm with larger pores greater than 200 gm were
fabricated. The scaffold pore architecture was highly in-
terconnected which is necessary for tissue ingrowth and
regeneration [54]. Based on their results from an animal
experiment, the interdisciplinary group proposed a scaf-
fold design concept which results in in vivo bone regen-
eration based on hematoma stabilization [56]. The
authors compare their in vivo bone engineering concept
to the induction phase of fracture healing. The osteop-
rogenitor cells which are in the blood of the osseous
wound are embedded in the scaffold microarchitecture
via the hematoma. The multipotent cells differentiate to
osteoblasts due to the presence of growth factors which
are released by the host bone. However, the emulsion Fig. 5. Schematicillustration of the fused deposition modeling(FDM)
freeze-drying method is user and technique sensitive. The process.
 The B i o m a t e r i a l s Silver J u b i l e e C o m p e n d i u m 183
D. W. Hutmacher/ Biomaterials 21 (2000) 2529-2543 2537

up to 9 5 % . At present, only p o r e sizes of up to 100 g m used to tissue e n g i n e e r b o n e a n d cartilage [62-64]. Excel-

can be r e p r o d u c i b l y f a b r i c a t e d by t h e r m a l l y i n d u c e d
phase-separation technology.
A n u m b e r of textile t e c h n o l o g i e s h a v e the p o t e n t i a l to
design a n d fabricate highly p o r o u s scaffolds [61]. Yet,
only so-called n o n - w o v e n mesh-like designs h a v e been
lent results in tissue e n g i n e e r i n g cartilage h a v e been
achieved by using n o n - w o v e n m e s h e s c o m p o s e d of poly-
m e r fibers of P G A , P G A / P D L A , a n d P G A / P L L A . This
w o r k has been reviewed by F r e e d et al. [65] a n d will n o t
be discussed here. In general, n o n - w o v e n c o n s t r u c t s can

Fig. 6. (a) 3D scaffold systems of various porosity and pore geometry fabricated by FDM. Magnification x 7.5, scale bar represents 1 mm. (i)-(iii)
lay-down pattern: 0/90~ nozzle tip: 0.016"; porosity: 50, 68, 75%; (iv)-(vi)0/90~ 0.010"; 50, 68, 75%; (vii)-(viii)0/60/120~ 0.016"; 68, 75%; (ix) 0/60/120~
0.010"; 80%; (x)-(xii) 0/60/120~ 0.010"; 50, 68, 75%. (b) Left: cross-sectional view of freeze-fractured PCL scaffold with lay-down pattern
0/72/144/36/108 ~ Right: plan view of same specimen. (c) Left: top view of PCL-HA scaffold with lay-down pattern 0/60/120 ~ The material
composition consists of 80% PCL and 20% HA by weight. Right: a close-up view of the same specimen at a higher magnification, shows that HA
particles are at the scaffold surface. (d) Freeze-fracture cross-sectional surface ( x 23) of a bioerodable scaffold designed for a bone/cartilage interface.
A 0/60/120 ~ and a 0/90 ~lay-down pattern of the roads shown in the upper and lower portion respectively of the SEM picture. Despite being fabricated
with different lay-down patterns, the porosity (67%) of both portions could be made identical. However, by applying FDM the porosity of each
portion of the 3D scaffold can be varied according to the road spacing for individual layers.
 184 The Biomaterials Silver Jubilee Compendium
2538 D.W. Hutmacher / Biomaterials 21 (2000) 2529-2543

Fig. 6. (Continued).

be only used for Strategy II since their physical proper-
ties do not allow load-bearing applications.
All the above-described technologies except the mem-
brane-lamination method, do not allow the fabrication of
a 3-D scaffold with a varying multiple layer design. Such
a matrix architecture is advantageous in instances where
tissue engineers want to grow a bi- or multiple tissue
interface, e.g. an articular cartilage/bone transplant.
Rapid prototyping technologies as well as so-called 'wa-
fer stacking systems' [66] (Fig. 4) have the potential to
design a 3-D construct in a multi-layer design within the
same gross architectural structure.
In engineering literature [67] Rapid prototyping Tech-
nologies (RP) also called Solid Free Form fabrication
(SFF) methods are defined as a set of manufacturing
processes that are capable of producing complex-free
 The Biomaterials Silver Jubilee Compendium 185
D.W. Hutmacher / Biomaterials 21 (2000) 2529-2543 2539

form parts directly from a computer-aided design

(CAD) model of an object without part specific toolin
g or knowledge. Unlike machining processes such as
milling, drilling, etc., which are subtractive in nature, RP
systems join together liquid, powder and sheet materials
to form parts. Layer by layer, RP machines fabricate
plastic, wood, ceramic and metal objects using thin hori-
zontal cross sections directly from a computer-generated
model.
Rapid prototyping technologies, such as 3-D printing
(3-DP) and fused deposition modeling (FDM) allow the
development of manufacturing processes to create por-
ous scaffolds that mimic the microstructure of living
tissue. Three-dimensional printing--developed at the
Massachusetts Institute of Technology [68]--is also
a rapid prototyping technology which has been used to
process bioresorbable scaffolds for tissue engineering ap-
plications [69,70]. The technology is based on the print-
ing of a binder through a print head nozzle onto
a powder bed, with no tooling required. The part is built
sequentially in layers. The binder is delivered to the
powder bed producing the first layer, the bed is then
lowered to a fixed distance, powder is deposited and
spread evenly across the bed, and a second layer is built.
This is repeated until the entire part, e.g. a porous scaf-
fold, is fabricated. Following treatment, the object is
retrieved from the powder bed and excess unbound pow-
der is removed. The speed, flow rate and even drop
position can be computer controlled to produce complex
3-D objects [63]. This printing technique permits CAD)
and custom-made fabrication of bioresorbable hybrid
scaffold systems. The entire process is performed under
room-temperature conditions. Hence, this technology
has great potential in tissue engineering applications.
Biological agents, such as cells, growth factors, etc., can
be incorporated into a porous scaffold without inactiva-
tion if non-toxic binders, e.g. water can be used [71].
Unfortunately, aliphatic polyesters can be only dissolved
in highly toxic solvents, such as chloroform, methylene
chloride, etc. To date, only bioresorbable scaffolds with-
out biological agents within the polymer matrix and in
combination with particle leaching have been processed
by 3-D printing. In addition, the mechanical properties
Fig. 7. Schematic illustration of the surgical placement of a tissue
and accuracy of the specimen manufactured by 3-D engineered a bone/cartilage interphase. The dental cylinder implant-
printing have to be significantly improved [72]. like design of the bony part of the scaffold allows to apply a press-fit
The FDM process forms 3-D objects from a CAD file between the host bone and the cylindrical portion of the scaffold.
as well as digital data produced by an imaging source
such as computer tomography (CT) or magnetic reson-
ance imaging (MRI) [73]. The process begins with the In effect, the process draws the designed model (scaffold)
design of a conceptual geometric model on a CAD work- one layer at a time.
station. The design is imported into a software, which Thermoplastic polymer material, 1.78 mm in diameter,
mathematically slices the conceptual model into horizon- feeds into the temperature-controlled FDM extrusion
tal layers. Toolpaths are generated before the data is head where it is heated to a semi-liquid state. The head
downloaded to the FDM hardware. The FDM extrusion extrudes and deposits the material in ultra-thin layers
head (Fig. 5) operates in the X and Y axes while the onto a fixture less base. The head directs the material
platform lowers in the Z-axis for each new layer to form. precisely into place. The material solidifies, laminating to
 186 The Biomaterials Silver Jubilee C o m p e n d i u m
2540 D.W. Hutmacher/ Biomaterials 21 (2000) 2529-2543

the preceding layer. Parts are fabricated in layers, where
a layer is built by extruding a small bead of material, or
road, in a particular lay-down pattern, such that the layer
is covered with the adjacent roads. After a layer is com-
pleted, the height of the extrusion head is increased and
the subsequent layers are built to construct the part. In
the past, non-medical and medical F D M users could
only use a few non-resorbable polymeric materials, such
as polyamide, ABS, and other resins. The broadening of
the research and application scope of F D M results in
a need/demand to evaluate the critical factors for using
the new polymeric and composite materials on F D M
systems [74].
At present, the author's multidisciplinary group has
been able to evaluate the parameters to process a number
of potential scaffold materials, such as P C L and
P C L / H A by F D M . To our knowledge, we are the first
group to report the processing of bioresorbable scaffolds

Fig. 8. (a) Schematic illustration of a perfusion culture chamber for tissue engineering a bone/cartilage interphase; (b) schematic sketch of the
bioreactor concept for the tissue engineering of bone and cartilage simultaneously, in the one device like scaffold architecture.
 The Biomaterials Silver Jubilee C o m p e n d i u m
D.W. Hutmacher/ Biomaterials 21 (2000) 2529-2543
(Fig. 6a-d) for tissue engineering applications using
FDM.
5. Tissue engineering of a articular cartilage-bone
interface
Articular cartilage repair, regeneration and generation
have been reviewed by Buckwalter and M a n k i n [75] as
well as N e w m a n [76]. Both the reports have concluded
that in the last two decades, clinical and basic scientific
investigations have shown that the i m p l a n t a t i o n of
artificial matrices, growth factors, perichondrium, and
periosteum, can stimulate the f o r m a t i o n of cartilaginous
tissue in o s t e o c h o n d r a l and chondral defects in synovial
fluids. However, the available evidence indicates that the
results vary considerably a m o n g the individuals, and that
the tissues formed using these t r e a t m e n t regimes do not
duplicate the composition, structure, and mechanical
properties of n o r m a l articular cartilage [77-81]. In addi-
tion, none of those matrix designs could be securely fixed
under load-bearing conditions to the o s t e o c h o n d r a l bone
which is a conditio sine qua non from a biomechanical
and clinical point of view.
The author's multidisciplinary g r o u p has concep-
tualized a rationale for tissue engineering a load-bearing
o s t e o c h o n d r a l t r a n s p l a n t (Fig. 7). The scaffold design,
material and fabrication technology, as well as the
bioreactor design (Fig. 8a and b), enables the tissue
engineering of bone and cartilage simultaneously, in the
one scaffold architecture. The device-like design of the
bony scaffold part allows a secure b o n e - t o - b o n e fixation
of the articular b o n e - c a r t i l a g e interface. The concept has
been discussed in detail elsewhere E82].
6. Conclusions
The application of regulatory a p p r o v e d biomaterials
to design and fabricate 3-D scaffolds has strongly
s u p p o r t e d the drive for the establishment of tissue
engineering research. Reviewing the experimental and
clinical studies, it can be concluded that the ideal
scaffold and matrix material for tissue engineering
bone and cartilage has not yet been developed. In
general, the tissue engineers do not i m p l e m e n t inno-
vative scaffold fixation features into their design concept.
However, from a clinical point of view, the secure
and user friendly transplant fixation is a conditio sine
qua non. The shape of a m e s e n c h y m a l tissue such as
a bone and articular cartilage is often critical to its
function. Ideally, a polymeric scaffold material should
permit application of a solid-free form fabrication tech-
nology, so that a porous scaffold with any desired 3-D
g e o m e t r y can be designed and fabricated by using CT
and M R I data.
187
2541
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