dental materi als 2 6 ( 2 0 1 0 ) e94e99

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Relationship between bond strength tests and other
in vitro phenomena
Junji Tagami
a,b,
, Toru Nikaido
a
, Masatoshi Nakajima
a
, Yasushi Shimada
a
a
Cariology and Operative Dentistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University,
1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8549, Japan
b
Global Center of Excellence Program, International Research Center for Molecular Science in Tooth and Bone Diseases,
Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8549, Japan
a r t i c l e i n f o
Article history:
Received 24 November 2009
Accepted 24 November 2009
Keywords:
Bond strength
Micro-leakage
Nanoleakage
Bond durability
Super-Dentin
a b s t r a c t
Objectives. The objective of this manuscript is to outline the relationship between bond
strengthtests of adhesive materials andother invitrophenomenabyreviewing the literature
generated over the thirty years.
Methods. Information was gathered from nearly 30 published articles appearing in dental
literature. Studies were predominantly identied through a search of the PubMED database
with a few additional studies published in Japanese domestic journals.
Results. Studies were included that provided evidence for the relationship between results
of evaluations of the bond strengths and other in vitro phenomena such as micro-
leakage, nanoleakage, structural of bonding interface and mechanical properties of bonding
interface. An attempt was made to select articles that spanned the timeframe from approx-
imately 1980 to today to try to ensure that the classic literature as well as the latest
information was included.
Conclusions. Previous studies have indicated bond strength did not correlate with the results
of micro-leakage tests nor gap formation at the cavity margin. The nanoleakage test and
morphological and chemical characteristics of the bonding interface had potential to predict
the dentin bond durability.
Signicance. Bond strength test results and other in vitro phenomena was not evenly corre-
lated. According to the development of the materials, the most appropriate bond test and
analysis of the in vitro phenomena should be also developed.
2009 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.
1. Introduction
Adhesive restorations are widely distributed as the routine
procedures inoperative and restorative treatments. The bond-
ing performance of the adhesive materials is evaluated by
means of the bond test to enamel and dentin. In general, ten-
sile bond test and shear bond test have been applied. Because
of the improvement of the bonding performance of the mate-

Corresponding author at: Cariology and Operative Dentistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental
University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8549, Japan.
E-mail address: tagami.ope@tmd.ac.jp (J. Tagami).
rials, the micro-tensile bond test [1] and micro-shear bond
test [2] are becoming more commonly applied for testing the
recent adhesive materials. The main purpose of bond tests
is recognized as the comparative evaluation of the bonding
performance of materials.
Micro-leakage tests at the margins of restorations have
beenwidelyperformed, since the micro-leakage of the restora-
tion had been believed to be the signicant factor to cause
post-operative tooth sensitivity, marginal discoloration and
0109-5641/$ see front matter 2009 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.dental.2009.11.156
dental materi als 2 6 ( 2 0 1 0 ) e94e99 e95
recurrent caries. In the micro-leakage tests, various cavity
designs and lling techniques have been evaluated. The sep-
aration of restoration from cavity walls occurs as the result of
contraction stress of composite resin during polymerization,
which is higher than that of the bond strength.
However, even when micro-leakage is not observed at the
interface between bonding material and dentin, a channel
that accepts the penetration of water and other solutions was
found at the bonding interface. This nano-sized space is now
recognized as the nanoleakage space [3]. The nanoleakage test
is often applied in recent papers to compare the dentin bond-
ing performance of materials.
Morphological analysis of the bonding interface is also
performed in most of the papers on dentin bonding. These
micro-structural studies are sometimes combined with acidic
and/or basic treatment. Through the studies on micro-
structure of dentin bonding interface, the so-called hybrid
layer was pointed out [4].
The evaluation of adhesive materials is carried out by the
methods described above in most of the recent studies. The
recent materials are proven to exhibit extremely high bond
strengths when they are tested after 24h of bonding. However,
the durability of bonding, especially dentin bonding, remains
a major issue for most of the adhesive materials. The bond-
ing interface consisted of multiple structures such as bonding
resin, hybrid layer and dentin substrate. The characteristics of
these components are becoming more important for predict-
ing the future degradation of bonding.
The above-mentioned phenomenon should be discussed
with association to each other. The bond strength of the mate-
rials, particularly dentin bonding, is discussed in the present
paper particularly correlating it with the other in vitro phe-
nomenon.
2. Micro-leakage
Micro-leakage has been evaluated using dye solutions to
detect the space between the restoration and cavity walls
and contraction gaps have been evaluated as the gap width
at the cavity margin [5,6]. Though the methods are not the
same, these two phenomenon cannot be differentiated from
each other. The gap and the micro-leakage space are the
results of the separation between the restoration and cav-
ity wall. And the separation occurs when the bond strength
to the cavity wall is lower than the contraction stress of the
restoration.
Whenthe mechanismfor debonding at cavitywalls is taken
into account, bond strength and micro-leakage are considered
to be correlated. Actually, the gap width was reported to be
correlated with the shear bond strength [5,6]. In these studies,
the cavity was prepared in human dentin, and its depth and
diameter were 1.5mm and 3mm, respectively. In contrast, a
correlation between bond strength and marginal adaptation
could not be demonstrated when class V cavities were lled
with various bonding materials, procedures, and lling mate-
rials [7]. In this study, the gap was evaluated as the fraction of
sample numbers showing a cervical gap, not by the gap width.
Even when the same adhesive procedure was applied, correla-
tion was found between the gap test and shear bond strengths
at 5minor 24hafter bonding. The fractionof samples showing
a cervical gap correlated inversely with the Youngs module
values of the restoratives used to ll the cavities. Also, the
application of an intermediate unlled resin layer improved
the quality of marginal adaptation.
Other studies have shown that the mechanical proper-
ties of dental composites were highly correlated with bond
strengths to dentin or enamel [811]. On the other hand, the
bond strength at the dentin cavity oor with a stiffer compos-
ite was reportedtobe more affectedby the cavity conguration
than that of a less stiff composite [11]. The stiffness of com-
posites is considered to inuence differently bond tests and
micro-leakage tests.
Thus, the results of micro-leakage tests are inuenced by
cavity size and shape, bonding procedures, lling procedures
andmaterials. The bondstrengthandmicro-leakage shouldbe
discussed separately, since the bond strengthdoes not directly
result in the micro-leakage score. However, the evaluations of
the micro-leakage and gap are benecial for the evaluation of
the restoration procedures including the adhesive materials
and procedures, and lling materials and procedures, simu-
lating the clinical application.
3. Nanoleakage
The recent adhesive materials provide excellent bonding to
both enamel and dentin. Especially dentin bonding has been
extremely improved and the bonding interface is observed
by SEM to be perfectly sealed. The dentin hybridization was
reportedto be very important for bonding. However, the hybrid
layer is not uniformly created, leaving the demineralized
dentin without the impregnation of adhesive resin materials.
This area is thought to accept the penetration of substances
that are smaller than the space in the hybrid layer. Nanoleak-
age may be recognized as a phenomenon, which provides the
space or defect at the bonding interface to accept the penetra-
tion of solutions, ions and enzymes.
Durability of dentin bonding is often discussed with the
recent materials, however, dentin bonding has been the most
signicant issue for any adhesive material. Kiyomura [12]
reported the drastic decrease of the dentin bonding of Super
Bond after 3 months, 2 years, and 4 years. The initial bonding
after 24h was 18MPa, however, the bond strengths decreased
to 8MPa at 3 months, and 4MPa at 2 years and 5 years after
bonding. The degradation of the bonding material, which
was the MMA with 4-META activated by TBBO catalyst, was
considered to be the main cause of the decrease of bond
strength. The drastic decrease of the bond after 2 years and
5 years was considered to be caused by the degradation of
the hybrid layer as well as the interface between the hybrid
layer and underlying dentin. Regarding the adhesive with
self-etching primer, comparatively more stable bond dura-
bility was conrmed than that with the adhesive system
using the more aggressive acid etching [13]. The fracture
tended to occur at the bottom of the hybrid layer with the
bonding system using the acid etching after 1 year storage,
however, the fracture surface was covered by the bonding
resin throughout the whole time period with the self-etching
system.
e96 dental materi als 2 6 ( 2 0 1 0 ) e94e99
Fig. 1 Micro-tensile bond strengths of Clearl Liner Bond
2V (LB2V), Fluoro Bond (FB), Single Bond and One Step at
each period. From the left, 1 day, 3 months, 6 months and 9
months, respectively.
Fig. 2 The area of silver penetration at the bonding
interface at each evaluation period with the 2-step
self-etching adhesives. L: Clearl Liner Bond 2V, F: Fluoro
Bond.
Thus, the bond durability, especially dentin bond durability
has been recognized to be strongly inuenced by the dura-
bility of the components at the bonding interface, such as
the hybrid layer and bonding resin layer. From this point of
view, the evaluation of the nanoleakage channel in the hybrid
layer is believed to be very effective to speculate on the bond
durability of the materials.
The nanoleakage was quantitatively evaluated with the
correlation to the micro-tensile bond strengths of various
adhesive materials at 1 day, 3 months, 6 months and 9 months
after bonding [14,15]. The materials tested were Clearl Liner
Bond 2V, Fluoro Bond, Single Bond and One Step. The results
ware shown in Figs. 13.
Fig. 3 The area of silver penetration at the bonding
interface at each evaluation period in the adhesive resins
with phosphoric acid etching.
For three adhesive systems, Clearl Liner Bond 2V (LB2V),
Single Bond (SB) and One Step (OS), the bond strengths
gradually decreased over time, although there were no sta-
tistically signicant differences in the Fluoro Bond (FB) bond
strength among the four time periods tested (p>0.05). Silver
penetration in specimens bonded with LB2V and FB gradually
increased over time. Regression analysis showed a higher
correlation between bond strength and silver penetration at
9 months for specimens bonded with LB2V (R
2
=0.844) than
at shorter time periods. It was speculated that hydrolytic
degradation within the hybrid layer gradually increased due
to water penetration through nanoleakage channels, resulting
in lower bond strengths and interfacial failure after as little
as 9 months.
The tensile bond strength of both materials gradually
decreased over time. Specimens bonded with OS showed less
silver nanoleakage at one day compared to 3, 6 and 9 months
(p<0.05), but there were no signicant differences betweenthe
nanoleakage measured at 3, 6 and 9 months. In contrast, for
specimens bonded with SB, there were no statistically signi-
cant differences inthe silver nanoleakage among the four time
periods tested (p>0.05). No correlation was observed between
bond strength and nanoleakage for either bonding system.
Nanoleakage occurred in both adhesive systems, and bond
strengths gradually decreased over time. However, there was
no correlation between bond strength and nanoleakage for
either adhesive system with phosphoric acid etching. This is
probably because the nanoleakage score of these adhesives
had already been comparatively higher for the specimens in
the 24h storage group.
Actually the adhesives with phosphoric acid etching
tended to exhibit more nanoleakage than the 2-step self-
etching adhesive [16]. Fig. 4 shows the bonding interface
between Single Bond and dentin. Clear silver penetration was
conrmed in the adhesive and hybrid layer under the FE-SEM
and EDS. On the other hand, the 2-step self-etching adhesive
exhibited much less silver penetration at the bonding inter-
face (Fig. 5).
As shownabove, the nanoleakage phenomenonis observed
remarkably with the bonding systems using phosphoric acid
etching. When the etching time was prolonged, nanoleakage
increased, however, the initial bonding of Single Bond was
not inuenced [17]. However, the nanoleakage is considered
to have a signicant relationship with the durability of dentin
bonding, i.e. the initial nanoleakage may be an indicator of the
durability of the dentin bonding.
4. Structural analysis of bonding interface
Numerous studies have reported on the structure at the bond-
ing interface, especially dentin and adhesives. A hybrid layer
was conrmed under the SEMwith acidic and basic treatment
[4], then the ultra-structure of the hybrid layer was revealed
with various treatments under the SEM and TEM [18,19]. The
structural analysis is still informative to discuss the bonding
performance, particularly the bond durability.
The recent studies revealed a dentin layer adjacent to the
hybrid layer, which shows more resistance against acidic and
basic attack than underlying normal dentin. This acidbase
dental materi als 2 6 ( 2 0 1 0 ) e94e99 e97
Figs. 4 (left nd 5 (right))}] FE-SEM image and EDS analysis of the bonding interface between dentin, and Single Bond (Fig. 4)
and Clearl SE Bond (Fig. 5).
resistant dentin is clearly distinguished from the so-called
hybrid layer [2022]. For disclosing the acidbase resistant
zone, the bonded specimens were placed in an articial dem-
ineralizing solution (pH 4.5) for 20min and in 5% NaOCl
for 20min. The specimens were sectioned, polished, and
argon-ion etched for 7min for SEM examination of the
dentinadhesive interface.
Fig. 6 shows the interface between Clearl SE Bond and
dentin after the acidbase challenge. The acidbase resistant
zone (ABRZ) is clearly observed between the hybrid layer and
dentin. Under the TEM, the ABRZ is also conrmed (Fig. 7),
however, in the case of the adhesives with phosphoric acid
etching, the ABRZ could not be formed (Fig. 8) [23]. As demon-
strated previously, the penetration of the bonding resin into
the phosphoric acid etched dentin was not sufcient, and
the effect of the adhesive material could not observed. Since
the ABRZ was more chemically and mechanically stable than
Fig. 6 Bonding interface between Clearl SE Bond and
dentin, after the acidbase challenge from the surface.
Acidbase resistant zone (ABRZ) is observed between
dentin (D) and hybrid layer (H).
normal dentin, it was termed as the Super-Dentin by the
authors [24]. The Super-Dentin is believed to be very effec-
tive for preventing recurrent caries around the margins of
restorations.
These observations may be discussed correlating with the
dentin bond durability of each material. Especially when the
self-etching material with uoride release was evaluated, the
thicker and more mineral dense ABRZ was observed than
that without uoride release. Also the uoride releasing adhe-
sive was reported to exhibit more dentin bond durability
than those without uoride release [25]. Though the mech-
anism of creating the ABRZ is not clear, the reinforcement
of the underlying dentin may inuence the dentin bond
durability.
The morphological research does not directly correlate
with the results of bond strength testing, since the bond
strength value is also regulated by the chemical reaction
between adhesive resin monomers and hydroxyapatite. The
chemical bonding ability and the stability of the salts are
reported to be very variable among the monomers [26,27]. Like
the bond test, the ultra-structure at the bond surface must be
the result of bonding inuenced by many factors. The chemi-
cal reaction of the monomers is thought to be reected in the
interfacial structure. The appropriate conditioning procedures
to disclose the differences of morphologies derived by these
factors should be developed.
5. Mechanical properties of bonding
interface
When the components at the bonding interface, such as lling
material, bonding resin, hybrid layer and underlying dentin
are bonded or connected strongly enough to each other, the
bondstrengthis determinedby the mechanical strengthof the
components. The weakest part should be fractured during the
e98 dental materi als 2 6 ( 2 0 1 0 ) e94e99
Figs. 7 (left nd 8 (right))}] TEMs of the bonding interface between dentin and Clearl SE Bond (Fig. 7) and Single Bond (Fig.
8).
testing. From this point of view, evaluation of the mechanical
strengths of interfacial components is considered to correlate
with the bond strength. The hardness and Youngs modulus of
the successive layers across a resindentin bonding area were
determined [28].
Recent adhesive resin materials, especially self-etching
adhesives that are categorized as one step adhesives, contain
hydrophilic monomers, water and other solvents. Because of
their components, the water absorption and solubility of the
bonding resin itself is considered to be a signicant factor
in maintaining the stable mechanical properties of the bond
layer [2931]. That must correlate with the bond durability,
as is already reported by many researchers. According to the
decrease of the mechanical properties of the bonding resin,
bond strength also decreased and more fracture was observed
at the bonding resin itself [13].
The situation must be the same when bonding to enamel.
Enamel bonding is also dependent on the mechanical
strengths of the components constituting the interface, as
well as the chemical bonding and micromechanical bonding
between the components. All these factors are extremely vari-
able among the materials.
6. Summary
The bond strengths of the adhesive resin materials are deter-
mined by many factors. The bonding among the components
of the interface, whichis obtained by chemical bonding and/or
micromechanical bonding, must be a signicant factor. The
mechanical strength of each component is also a very signi-
cant factor to inuence the bond strength. Eachadhesive resin
material has its own composition of adhesive resin monomer,
catalyst, other resin monomers and solvent. Phosphoric acid
etching and the self-etching technology are extremely dif-
ferent. As the in vitro studies of bonding materials, the
most appropriate measure should be selected to evaluate and
compare the performance of the materials, preferably with
consideration for the clinical situation. It must be benecial
also for improving the materials.
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