Research Article

Physical properties of the dentin-enamel junction region

ISAO URABE, DDS, MASATOSHI NAKAJIMA, DOS, PHD, HIDEHIKO SANO, DDS, PHD & JUNJI TAGAMI, DDS, PHD

ABSTRACT: Purpose: To determine the physical properties between enamel and dentin at the dentin-enamel junction
(DEJ) region of natural tooth structure. Materials and Methods: Ultimate tensile cohesive strengths of the DEJ region
of human and bovine teeth were measured using a microtensile test, and the nanohardness and Young's modulus from
deep enamel to superficial dentin of human teeth were measured using a nanoindentation tester. Results: Tbe mean
ultimate tensile cohesive strengths of bovine and human DEJ region were 47.7 MPa and 51.5 MPa, respectively. The
nanohardness, dynamic hardness and Young's modulus of the DEJ region showed moderate values between those of
enamel and dentin. Comparison of the cohesive strength of the human DEJ region with previous results of the bond
strength of resin bonding systems indicate that current resin bonding systems might reproduce the biological adhesion
of enamel to dentin in terms of the tensile strength. Moreover, the indentation properties of the DEJ region showed
higher values than those of the underlying dentin or of resin impregnated dentin. (Am J Dent 2000;13:129-135).

CLINICAL SIGNIFICANCE: The dentin-enamel (DEJ), which contains intermingled enamel and dentin, proved to be a
region that resists well to tensile stress and has intermediate hardness and elastic modulus between enamel and dentin.
Novel resin bonding systems have high tensile bond strength as well as tensile cohesive strength of the DEJ region.
Accordingly, these materials have sufficient bonding performance regarding bond strength. New principles for resto-
ration supported by adequate bond strength should be established.
CORRESPONDENCE: Dr. Isao Urabe, Cariology and Operative Dentistry, Department of Restorative Sciences, Graduate
School, Tokyo Medica} and Dental University, 1-5-4S, Yushima, Bunkyo-ku, Tokyo 113-8549, Japan. Fax: 81-3-5803-
0195. E-mail: urabe.opr@tmd.ac.jp

lntroduction The dentin-enamel junction (DEJ) is a unique biological

Improved resin bonding systems had succeeded in
adhering very strongly both to enamel and dentin. The
excellent sealing ability of resins bonded to acid-etched
enamel are well-known.1,2 The resin-dentin seal is much less
· rare to see enamel separate from dentin. lt has long been
uniform and the perfect resin-dentin bond continues to be a
challenge. Several investigators have suggested that there is
an inverse relationship between dentin bond strength and
microleakage using relatively large specimens.3,4 That is, at
bond strengths above 17-18 MPa measured in large bonded
specimens, there is little gap formation, while at lower bond
strengths larger gaps develop. Presumably, the high bond
strengths oppose the forces of polymerization contraction. 5,6
There may be an analogous situation between tensile bond
strength by the method of microtensile test7 and ideal dentin
bonding.
In the clinical situation, a new concept for the restoration
using these adhesive resin materials has been proposed and
applied for routine dental treatment. These principles of
cavity design are different from the principles established by
Black,8 since they presupposed that the non-adhesive re-
storative material could not bond to tooth substance ade-
quately. For example; in the case of adhesive restorations,
retention and resistance forms are not required for cavity
design. However, there is no standard for adequate bonding of
resin bonding systems in order to apply the principles for
adhesive restorations.
For the assurance of better adhesive restoration, it is im-
portant to examine the strength and structure of the teeth.
When considering the strength of the complex, including
restoration and the tooth substances, the strength of tooth
structures can be the true measure of the properties of restored
teeth.
interface between very hard, stiff enamel and much softer,
elastic dentin. During function, the more brittle enamel tends
to develop cracks, some of which extend through the full
thickness of enamel to the DEJ. However, clinically it is very
suspected that the structure of the DEJ may protect the tooth
from catastrophic fracture of enamel from dentin. Recently,
Lin & Douglas9 measured the fracture toughness of the DEJ
and suggested that structures on either side of the DEJ may
dissipate stress inhibiting further crack propagation. If the
resin bonding materials could adhere to dentin as well as the
enamel adheres to dentin in the DEJ region, the performance
of the resin bonding systems would be sufficient with regard
to the bond strength.
Therefore, to develop a better understanding of the
mechanical properties of the DEJ, this study measured the
ultimate tensile strength, the nanohardness and the Young's
modulus of the DEJ of bovine and human teeth.
Materials and Methods
Microtensile strength of the DEJ region - Extracted human
unerupted third molars and bovine incisors from mature
animals were used. After extraction, they were stored at 4ºC
in isotonic saline solution containing 0.2% sodium azide to
inhibit microbial growth.
Human teeth - The teeth were sectioned parallel to their long
axis into l mm thick slabs by means of a microtome saw
(Leitz 1600ª) (Fig. 1). The area of interest was the buccal
surface at its greatest height of contour. The pulpal side of
dentin was reduced by means of a steel round bur, and the
peripheral enamel was sanded with 600-grit waterproof
silicon carbide paper. Then, both the enamel and dentin sides
were acid-etched with 37% phosphoric acid (K-etchantb) for
 American Journal of Dentistry, Vol. 13, No. 3, June, 2000
130 Urabe et al

CJ
e
..l
'
Tlmm
human 3rd molar bovine incisor

A D trimming

1mm

dentin enamel
B E tensile test

t cyanoacrylate
cement

resin composite
! 1 •··-·- .,,

Fig. 1. Regional location of interest and the method of sample preparation for tensile testing. A: Block containing
enamel and dentin cut from human third molar and bovine incisor. B: Both enamel and dentin sides covered with
resin composite. C: Preparation of series slices approximately 1 mm thick. D: Specimens trimmed with diamond
burs approximately 1 mm wide at its narrowest dimensions. E: Trimmed specimens fixed to stainless steel grips with
cyanoacrylate cement.

Bovine teeth - Bovine incisors were prepared exactly like the

human teeth. The area of interest was the mid-coronal portion
of the labial enamel (Fig. 1).
l•
Testing - The apparatus that was used to measure the micro-
-1 1- tensile strength was a Bencor-Multi-T" testing apparatus in a

HII e
A B
D
Fig. 2. Regional location of interest and the method of sample preparation for
nanoindentation testing. A: Coronal portion of human third molar. B: 2 mm
thick slabs were prepared longitudinally. C: Slabs embedded in epoxy resin
and polished to a mirror-like surface. D: The cylinder was fixed with
modeling compound to set the polished surface parallel to the slide glass in
preparation for testing.
40 s, rinsed with water, bonded with a light-cured adhesive
bonding system (Clearfil PhotoBondb), and then covered with
a resin-based composite (Clearfil AP-Xb). Approximately 1
mm thick slices (Fig. 1) were made through the block
perpendicular to the interface of dentin and enamel (DEJ).
These slices were then trimmed and shaped, by means of
Super-fine diamond burs (c-16ff) in a high-speed handpiece,
into a gentle curve along the DEJ from both sides, (Fig. 1) to
form a square cross-section, approximately 1 mm wide at its
narrowest dimensions.
universal testing machine (Model AGS00Bc). The widest
portions of the specimens were fixed to flat stainless steel
"grips" by use of a small amount of cyanoacrylate cement
(Zapitb), with care taken to avoid any spread of cyanoacrylate
onto the test region of the specimen (Fig. l ). Tensile strength
testing was carried out at a crosshead speed of 1 mm/min. The
apparent ultimate tensile strength of the specimens was
calculated by dividing the load at failure by the cross-
sectional area of the specimen, and expressed in MPa. After
calculation of descriptive statistics, comparisons of the
differences between the ultimate tensile strength of human
and bovine teeth were tested for statistical significance using
Student's unpaired t-test.
SEM observation of the fractured surfaces - After measure-
ments of the ultimate tensile strength were made, the speci-
mens of human teeth were mounted on SEM stubs, dried
overnight in a desiccator, and then coated with gold. Then,
SEM observation of the fractured surface was carried out to
investigate where the failure occurred and to try to discern the
path of the fracture plane.
Nanohardness and Young modulus measurements - The
procedures of sample preparation for a nanoindentation test-
ing are shown in Fig. 2. Extracted unerupted human third
molars were used. Discs about 2 mm thick were cut in cross
section from the mid-coronal portion and then embedded in
American Journal of Dentistry, Vol. 13, No. 3, June, 2000
Electoromagnetic coil
Base
Motor
Motor
Base
l
Fig. 3. Principles of the nanoindentation tester.
epoxy resin (Epon 815B ) . After 24 hrs of polymerization
of the resin, the surfaces were polished with waterproof
silicon carbide papers (grit #600, 800, 1000) and diamond
pastes (particle size 6, 3, 1, 0.25 µm). Sample molds
were set on heated modeling compound, which served to
stabilize the specimen surface and orient them parallel to
the stage of the nanoindentation tester (ENT-11OOh). Ali
measurements were made on dry specimens.
The instrument used for this experiment was a depth
sensing computer controlled instrument with a three-sided
pyramidal diamond probe, as detailed in Figure 3.
The instrument was in an isolation enclosure with a tem-
perature controller on an ALD anti-vibration isolator in or-
der to prevent from influences of environmental conditions
such as the room temperature, floor-vibration and noise. Z-
axis displacement is measured by a capacitive
displacement. Loading control system was powered by
electromagnetic force and minimum load is 10 mgf. The
position of indentation can be programmed observing with
a CCD camera and 100 positions can be measured in each
measurement. For the precise measurement, the interval of
each indentation should be five times the size of
indentation to avoid the corruption of abutment.
Dentin-enamel junction 131
Sample stage
XYZstage
J
After setting a sample on the stage of the testing machine,
the instrument was programmed to create the position of
indentations. There were 10 indentations at 5 µm intervals
across the DEJ from deep enamel to mantle dentin in the
middle third of the buccal surface (Fig. 4). The indentations in
dentin were set on the intertubular dentin. The load on the
indenter was 400 mgf. After the performing the indentation,
the nanohardness and Young's modulus were calculated by an
attached computer.
Fig. 5 shows a load vs. displacement curve in the meas-
urement process. Values of nanohardness (NH), dynamic
hardness (DH) and Young's modulus (E*) were calculated
according to equations 1, 2 and 3, respectively:
NH = 3.7926 x 10 -2 p/h22 (1)
OH= 3.7926 x 10 -2 p/h1 2 (2)
E*= 1.8129 x 10 -3 1/h1 dP/dh (3)
where p is applied load, h1 is maximum (plastic) displace-
ment (µm), h2 is the permanent deformation depth (µm) and
dP/dh is the tangential slope at the maximum load in
unloading segment (Fig. 5). Each value was expressed in
kg/mm2 which was then converted to GPa. Indentation tests
were carried out on six specimens. After the indentation tests,
the geometry of the indentation marks were confirmed using a
laser scanning microscope (1LM15W i).
 American Journal of Dentistry, Vol. 13, No. J, June, 2000
132 Urabe et al

Enamel Dentin
:::::::::::::::::::::::::::::::::::::::::::::: :::::::::::::::::::::::

lOµ r:fl
j \ j j \ \ij jij j j j \ j \ \ j j \i j j j j j j j j j jj \{
,, '': : : : :-: -:-: -:-:-:-: : : :-:- -::- -: :- :-:-:- -::--: :-:-:-:-:-:-:-:-:
::. :.:::. y. :.:.:::. . :.::. :::::. ...............
y·.:.::::::::: :::::.y.:::: ::::!
#4 .· .·.·.·.·.·.·.· .·. ·.·.·.·.·.·.·.·.·.·.· .·.·.·.·.·.·.·. ·.·.
:-:.:-:-:-:-:-:-:->:-:-:.:-:

12H JIT(/ \ \Mi 3

◆ : - :- : - :- : - : - : - : ♦ ♦ :- :- : - : - : - : -: ♦ :- :- : - : - :-

#1 #3

> JU: } \/{ : : :

DEJ
Fig. 4. Position of indentations.

Table l. Comparison of tensile strength of bovine and human DEJ (MPa).

p Bovine Human

Mean 47.7 51.5

unloading Standard deviation 10.8 13.0
Minimum 29.2 37.8
Maximum 62.9 73.2
Load Number of specimens 10 10

Table 2. Mean values of hardness and Young's modulus of enamel, DEJ area
and dentin (GPa).

Enamel DEJ Dentin

h2 h1
Displacement Nanohardness 4.48 2.37 0.70
Dynamic hardness 2.90 1.58 O.55
fig. 5. Schematic representation of a load vs. displacement curve. Young's modulus 87.68 53.18 24.04

Results
Microtensile strength of the DEJ region - The means and
standard deviations of the microtensile cohesive strengths of
bovine and human DEJ region are given in Table l. The
microtensile strengths of bovine and human teeth were 4 7.7
MPa and 51.5 MPa, respectively. There was no statistically
significant difference between the ultimate tensile strength of
the two groups (P> 0.05).
SEM observation of fractured surfaces - All 10 specimens
fractured in the vicinity of the DEJ. Three types of fracture
patterns were observed as illustrated in Fig. 6. Five of these
specimens fractured 100% in dentin, and the fracture points
were about 10 µm below from DEJ perpendicular to the ori-
entation of the dentin tubules (Figs. 6A, 7). Only one speci-
men fractured 100% in enamel, and the fracture surface was
uneven, but seemed to follow the orientation of the enamel
rods (Figs. 6C, 8).
Four specimens fractured both in dentin and enamel, and
the failures occurred only in the DEJ area. Furthermore, the
fracture surfaces were observed to extend from enamel into
dentin along the orientations of enamel rods and dentin tu-
bules at the DEJ, and the crack propagation in dentin side,
however tended to be parallel to the DEJ (Figs. 6B, 9).
Nanohardness and Young's modulus measurements - Fig. 10
shows successive static and dynamic hardness values meas-
ured on either side of the DEJ. The mean values of nano-
hardness in enamel and dentin area were 4.48 ± 0.44 and 0.70
± 0.12 GPa, respectively (Table 2). The mean values for
dynamic hardness in enamel and dentin area were 2.90 ± 0.23
and 0.55 ± 0.09 GPa, respectively (Table 2).
The mean values for Young's modulus of elasticity in
enamel and dentin area were 87.68 ± 5.94 GPa and 24.04 ±
3.92 GPa, respectively (Fig. 11). The mean depth of the
indentations of enamel, DEJ region and dentin were 0.22,
0.30 and 0.51 µm respectively. All of these values indicate
that the DEJ region has intermediate hardness and elastic
modulus between those of enamel and dentin (Table 2).
Discussion
Microscopically, the DEJ is not a two-dimensional inter-
phase but is a highly complex, three-dimensional interface
about 20-50 µm wide10 resting on a unique form of dentin
called mantle dentin. 11 The dentin side of the DEJ is com-
posed of a series of concavities with average diameters of
about 20 µm.12 The borders of these concavities form circular
ridges that rise up about 10 µm and are about 1-2 µm wide.
The enamel surface forms a series of convexities that extend
 American Journal of Dentistry, Vol. 13. No. 3. June, 2000
Enamel
DEJ region
/(\
1
Dentin
A B
Fig. 6. Fracture patterns in microtensile test. A: in dentin. B: in both enamel and dentin C: in enamel.
Fig. 7. Pattern of fracture of DEJ specimens in dentin, but parallel to DEJ. SEM x 1500. Fig.
Fig. 8. Failure within enamel during tensile testing. SEM x1500.
Fig. 9. Complex fracture pattern through both enamel and dentin. (D: Dentin. E: Enamel.)
into the dentin concavities.12 Further complicating this
structure is the presence of enamel tufts that periodically
extend from the enamel convexities up into the enamel for a
distance of 50-100 µm in a fan-like shape. Enamel tufts are
hypomineralized regions of retained enamel proteins. Less
frequent are the enamel spindles which represent odontoblasts
which forced their way into developing enamel, especially
over cusps tips.10 Finally, there are enamel lamellae which
are protein-filled defects that extend variable distances from
the enamel surface, sometimes reaching the DEJ. These are
regarded as developmental defects but are difficult to
distinguish from cracks that develop in enamel with function.
At the junction between enamel and dentin, there is a
change in apatite crystal size and orientation.14 In enamel, the
apatite crystallites tend to be rather large and are bonded to
their neighboring prismatic crystals or to interprismatic
protein-crystal complexes.14 In dentin, the apatite crystals are
much smaller15,16 and are bound to each other or to collagen
fibrils.17 There is an intermingling of the two apatite crystal
types at the DEJ. 17,18 The bond energies of these crystal-to-
Dentin-enamel junction 133
. .·..•·.····...........··
······.. .,........ ...\ ... ...·....···.·.·..._/·..../ ·!
a pprox.
:[, ...',-.:.;.l".1-1-:1:1
= ·.n,i .
10 µm
e
.)./.;r-J?;!=?!L
- fi,: :J ít.
crystal or crystal-to-collagen associations are not known but
must contribute to the cohesive strength of the structure.
Previous studies on the physical properties of tooth
structure have reported that the cohesive strength of enamel in
tension is 10-20 MPa,19,20 and that of dentin is 94-105 MPa
21,22
using a microtensile test. The same microtensile test was
applied in this study, because it is suitable to evaluate the
tensile strength of a limited area such as DEJ. Supposedly, in
the comparison of these data, the ultimate tensile strength of
human DEJ (51.5 MPa) is higher than enamel but lower than
dentin. As for the cohesive strength of the DEJ, Pioch &
Staehle23 reported that the ultimate shear strength of the DEJ
was 39 MPa comparable to that of dentin. Besides, using a
micropunch method to measure the shear strength, Smith &
Cooper24 reported shear strength of 93 MPa in enamel, 138
MPa at the DEJ and 132 MPa in superficial dentin.
SEM observations revealed that the fracture never occurred in
the interface between enamel and dentin. That is, the direction
of crack propagation was always across the DEJ but never
within it. Similar fractures occurred in a previous study of the
shear strength of the DEJ23 in human teeth.

134 Urabe et al
0Pa
6
Dynamic lwdness
Nanohardnesa
4
t··········!·········+·' ····· f..\\\
OL-...J..---!:--5-;, O -¿- 1- -}t i:W-::m
20 15 10 10
Enamel Dcntin
Distance from lbc DEJ
Fig. 10. Nanohardness and dynamic hardness values measured across the
dentin-enamel junction.
One reason for the lack of interfacial failure of the DEJ
can be attributed to the complexity of the DEJ structure and
how that structure may modify crack propagation. The inter-
face between enamel and dentin is not a distinct two-dimen-
sional structure but is actually a three-dimensional interphase
or zone, which has a scalloped shape with a wave-like form.
In addition, there are enamel spindles, lamellae and tufts
running across the DEJ. These findings indicated that the
region around the interface of enamel and dentin has an un-
usual and unique structure. Whittaker 12 reported that the DEJ
was more scalloped in the occlusal portion of molars and that
it became less scalloped on the buccal region and was almost
flat in the cervical region. Goel et al25 modeled the DEJ by
DEJ by finite element analysis to determine the influence of
occlusal forces of the stress distribution of the DEJ. They
found that the highest stress concentrations in the DEJ
occurred at the cervical-third where the DEJ is relatively
nonscalloped12 and where the enamel is very thin.
In this investigation, we could not measure the exact bond
strength of the interface between enamel and dentin because
most of the failures occurred in very superficial dentin rather
than within the DEJ. However, the results indicate that the
tensile bond strength of dentin to enamel is above 50 MPa.
Namely, DEJ might be thought to be a strong junctional
region not easily broken.
Our values for the Young's modulus of dentin (24.0 ± 3.9
GPa) and enamel (87.7 ± 5.9 GPa) are similar to those
reported by others.26-31The value of nanohardness and
Young's modulus of enamel within 5 µm of the DEJ were
equal to or higher than those of dentin. One possible expla-
nation of this high stiffness of the DEJ is that the crystals of
hydroxyapatite from dentin and enamel are intermingled in
this area.10,12.18,32 Beyond that, Lin.& Douglas 32 reported that
the DEJ can be regarded as a fibril-reinforced bond which is
less highly mineralized than either enamel or bulk dentin.
They note that collagen fibers at the junctional area of the
DEJ are oriented at right angle to the mean plane of the
scalloping, and link enamel to dentin. Similarly, Baume33
stated that the orientation of collagen fibers in mantle dentin
is not random but oriented at right angles to the DEJ and that
American Journal of Dentistry, Vol. 13, No. 3, June, 2000
0Pa
1()() ...
80
f··-··+·-...t ···· t
60
40
I···········,...........¡...........¡ ............
20
1S 20
o 20 15 10
Enlmcl
5 O 5 10
Dentin
15 20 25
um
Dislance from lbc DEJ
Fig. 11. Young's modulus values measured across the dentin-enamel junction.
the collagen fibrils are aggregated into large (ca. 5 µm) col-
lagen fibers that extend into enamel. The collagen fibers in
the vicinity of DEJ were considered to serve to blunt or de-
flect crack propagation, thereby increasing the fracture
toughness of the DEJ.9,34
It is of interest to compare the tensile bond strengths of the
newest resin bonding system with the cohesive strength of
human DEJ region. Pereira et al 35 reported that the mi-
crotensile strength of Clearfil Liner Bond IIb and One-Stepj
adhesive resins to dentin are about 42 MPa and 48 MPa,
respectively. In addition, it was reported that the microtensile
strength of the prototype of Clearfil Liner Bond 2V (KB-
1300b) to human dentin was approximately 58 MPa.36 These
values can be directly compared with the results of the present
study, because the testing method (i.e., the microtensile test)
was the same among the papers. In terms of their ultimate
tensile strength, it might be expected that these resin bonding
systems can reproduce the tensile properties of the DEJ.
Besides, it is noteworthy that the failure mode of the DEJ was
different from that of resin-dentin or resin-enamel bonding
systems, which occurred mainly at the adhesive interface
between resin and tooth substrates.
The stiffness of these resins is probably similar to that of
other bonding systems, which vary from 0.8-2 GPa22, which is
much lower than that of enamel or dentin. Van Meerbeek et
al27 reported that the Young's modulus of the hybrid layer
between resin and dentin was lower than that of dentin. In
contrast, the stiffness of the dentin-enamel junction area
measured in the current study showed moderate values for the
Young's modulus between enamel and dentin. Therefore, it is
possible that the interface between the dentin and the resin
bonding system is less stiff than the DEJ. These factors are
considered to be the reason for the difference in fracture
pattern between bonding and DEJ.
It is still unknown if the difference of fracture pattern
comparing resin-dentin interface and DEJ is advantageous or
disadvantageous. However, regarding tensile strength, recent
resin bonding systems can be considered to bond efficiently to
dentin.
From these results, it can be concluded that the DEJ serves
as an intermediate structure that resists tensile stress well. As
American Journal of Dentistry, Vol. 13, No . 3, June.2000
mentioned above, the bond strength of resin bonding systems
to dentin seemed to be comparable to that between enamel
and dentin. Namely, bond strength of the resin bonding
systems were equal to the cohesive strength of the DEJ
structure, which has long been thought to protect the tooth
from the catastrophic fracture of brittle enamel from dentin.
If the resin bonding systems adhere to tooth substrate suf-
ficiently, the filled resin-based composite (RBC) and tooth
structure can be considered as a monolithic structure. Ausiello
et al37 investigated fracture resistance of endodontically-
treated premolars, adhesively restored with various materials.
They noted that the fracture resistance of endodontically
treated teeth when restored with RBCs and dentin bonding
systems was not significantly different from that of sound
teeth.
a. Leica Instruments, Heidelberg, Germany.
b. Kuraray Co., Ltd., Osaka, Japan.
c. GC Ltd., Tokyo, Japan.
d. Danville Engineering lnc., Danville, CA, USA.
e. Shimadzu Corp., Kyoto, Japan.
f. DVA Corp., Corona, CA, USA.
g. Nisshin EM Co., Ltd., Tokyo, Japan.
h. Elionix Corp., Tokyo, Japan.
i. Lasertec, Yokohama, Japan.
j. Siseo lnc., Schaumburg, IL, USA.
Dr. Urabe is on the hospital staff, Dr. Nakajima is Instructor and Dr. Tagami
is Professor and Chairman, Cariology and Operative Dentistry, Department of
Restorative Sciences, Graduate School, Tokyo Medical and Dental
University, Tokyo, Japan. Dr. Sano is Professor and Chairman, Department
of Operative Dentistry, Hokkaido University Dental School , Hokkaido,
Japan.
Ackno wledgements The authors wish to express their sincere gratitude to Dr.
D.H. Pashley, Medical College of Georgia, for discussions and encour-
agement. The authors also thank Mr. Hideo Suzuki and Mr. Takahiko
Uematsu of Elionix, lnc. for technical assistance in the operation of the
nanoindentation tester.
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