Dentin Bonding Durability of Two-step Self-etch Adhesives

with Improved of Degree of Conversion of Adhesive Resins

Kento Satoa / Keiichi Hosakab / Masahiro Takahashic / Masaomi Ikedad /Fucong Tiane /
Wataru Komadaf / Masatoshi Nakajimag / Richard Foxtonh / Yoshihiro Nishitanii /David H. Pashleyj /
Junji Tagamik

Purpose: To evaluate (1) the initial and long-term microtensile bond strengths of two-step self-etch adhesives with
different degrees of conversion (DC); (2) the elastic modulus of the respective adhesive resins; (3) the water sorp-
tion of the respective adhesive resins.
Materials and Methods: Two two-step self-etch adhesives, Clearfil SE Bond (CSE) and Clearfil SE Bond 2 (CSE2)
were used in this study. The DC was determined using ATR/FT-IR with a time-based spectrum analysis. Midcoronal
flat dentin surfaces of 24 human molars were prepared with 600-grit SiC paper for microtensile bond strength
(μTBS) testing. CSE and CSE2 were applied to the dentin surfaces according to the manufacturer’s instructions,
followed by composite buildups. The μTBS was measured after water storage for 24 h, 6 months, and 1 year. The
elastic modulus (before and after 1 month of water immersion) was determined by the three-point flexural bending
test and water sorption values by the water sorption test.
Results: CSE2 showed significantly higher DC than CSE. The μTBS of CSE2 was significantly higher than that of CSE
in all water storage periods. One-year water storage decreased the μTBS of CSE; however, it did not decrease that
of CSE2. Regarding the polymerized adhesive resins, the elastic modulus of CSE2 was significantly higher than that
of CSE before and after water immersion (p < 0.001), and the water sorption of CSE was higher than that of CSE2.
Conclusions: The higher DC of adhesive resins of two-step self-etch adhesives resists water aging and improves
the initial bond strengths and durability of the resin-dentin bond.
Keywords: two-step self-etch adhesive, degree of conversion, microtensile bond strength, water sorption, elastic
modulus.

J Adhes Dent 2017; 19: 31–37. Submitted for publication: 01.07.16; accepted for publication: 05.11.16
doi: 10.3290/j.jad.a37726

a g Junior Associate Professor, Cariology and Operative Dentistry, Department of

Graduate Student, Cariology and Operative Dentistry, Department of Oral
Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo
Medical and Dental University, Tokyo, Japan. Experimental design, performed
the experiments, wrote the manuscript.
b h
Assistant Professor, Cariology and Operative Dentistry, Department of Oral
Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo
Medical and Dental University, Tokyo, Japan. Experimental design, performed
the experiments, wrote the manuscript.
c Clinical Fellow, Cariology and Operative Dentistry, Department of Oral Health
Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical
and Dental University, Tokyo, Japan. Consulted on and performed statistical
evaluation, contributed substantially to discussion.
d Junior Associate Professor, Department of Oral Prosthetic Engineering, Gradu-
ate School, Tokyo Medical and Dental University, Tokyo, Japan. Consulted on
and performed statistical evaluation.
e Assistant Professor, Department of Cariology and Endodontology, Peking Uni-
versity School and Hospital of Stomatology, Beijing, China. Contributed sub-
stantially to discussion.
f Junior Associate Professor, Fixed Prosthodontics Unit, Department of Restora-
tive Sciences, Division of Oral Health Sciences, Graduate School, Tokyo
Medical and Dental University, Tokyo, Japan. Performed the experiments, con-
tributed substantially to discussion.
Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo
Medical and Dental University, Tokyo, Japan. Experimental design, co-wrote
and proofread the manuscript, contributed substantially to discussion.
Clinical Senior Lecturer, Department of Restorative Dentistry, King’s College
London Dental Institute, King’s College London, London Bridge, London, UK.
Proofread the manuscript, contributed substantially to discussion.
i Professor and Chair, Department of Restorative Dentistry and Endodontology,
Medical and Dental Sciences School, Kagoshima University, Kagoshima,
Japan. Proofread the manuscript, contributed substantially to discussion.
j Regents’ Professor, Department of Oral Biology, College of Dental Medicine,
Georgia Regents University, Augusta, Georgia, USA. Co-wrote and proofread
the manuscript, contributed substantially to discussion.
k Professor, Cariology and Operative Dentistry, Department of Oral Health
Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical
and Dental University, Tokyo, Japan. Idea, hypothesis, study design, wrote the
manuscript.
Correspondence: Assistant Professor Keiichi Hosaka, Cariology and Operative
Dentistry, Department of Oral Health Sciences, Graduate School of Medical and
Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-
ku, Tokyo 113-8549, Japan. Tel: +81-3-5803-5483; e-mail: hosaka.ope@tmd.ac.jp

Vol 19, No 1, 2017 31

Sato et al
C urrently, the self-etch adhesives (SEAs) are widely used
in clinical practice.1,24 SEAs are based on the use of
acidic functional monomers that can simultaneously demin-
eralize and prime dentin. Self-etch adhesives eliminate the
rinsing phase, which not only lessens clinical application
time, but also significantly reduces the technique sensitivity
and risk of making errors during application.4 SEAs can be
classified into two-step adhesives (2-SEAs) and one-step
adhesives (1-SEAs). Previous in vitro studies have reported
that 2-SEAs show higher bond strengths to tooth tissues
than do 1-SEAs.12,27 Two-step SEAs, which involve the ap-
plication of an additional layer of solvent-free hydrophobic
resin, create stronger adhesive layers than do 1-SEAs,
which contain hydrophilic monomers, water, and volatile sol-
vents.20
However, even relatively hydrophobic adhesive resins of
2-SEAs absorb water. This decreases their mechanical prop-
erties,14,15 because water can penetrate into nanometer-
sized spaces between polymer chains or cluster around
polar functional groups that are capable of hydrogen bond-
ing, due to its small molecular size and high molar concen-
tration.30 Extrinsic and intrinsic water diffusion into the ad-
hesive layer and the resin-dentin interface hastens
hydrolytic degradation of resin components within the hybrid
layer and/or adhesive layer and leads to plasticization of
the adhesive,13,34 loss of interfacial adhesive-dentin bond
strengths,31 and hydrolytic degradation in the resin-dentin
interface.20 In particular, the relatively thick adhesive layer
of 2-SEAs is potentially susceptible to extrinsic water ab-
sorption in the restorations.32 The adhesive layer overlying
the hybrid layer may help preserve the integrity of the hy-
bridized dentin by sealing it from water,1,14 and by acting as
a stress-absorbing layer. Therefore, if the adhesive layer
that couples composite to hybridized dentin becomes less
stiff over time by water absorption, it may adversely affect
stress distribution across the bonded interface, possibly
resulting in either plastic deformation or debonding under
repeated loading,18,23 which results in inferior durability of
the resin-dentin bond. As the mechanical behavior of
2-SEAs exposed to water for extended periods of time is
affected,8 improved mechanical properties of the adhesive
layer are necessary to ensure longevity of the resin-dentin
bond.13,14
Since it is generally accepted that better mechanical
properties have been attributed to higher percentages of
monomer-to-polymer conversion in resin-based materials,2,9
the degree of conversion (DC) of monomer into polymer
(percentage of C=C bonds becoming C-C bonds as part of
the polymer chain) plays an important role in the strength
and stability of dentin bonding.5 Additionally, incomplete
polymerization of adhesive resins results in enhanced water
permeability of the adhesive layer and can accelerate hydro-
lysis, leading to bond deterioration.11 Therefore, it is antici-
pated that a higher DC of the adhesive resin may lead to a
more desirable load transfer across the bonded interface
over time and superior resin-dentin bond durabil-
ity.9,17,19,28,33 To improve the polymerization efficacy and
increase the DC of the adhesive resins, previous studies
32
have examined the effects of direct/indirect light inten-
sity,28 light-curing distance,39 light-curing time,39 and types
of light-curing units.7,39 On the other hand, optimizing the
ingredients of adhesives is another approach, because the
DC of adhesive resins depends on photo-initiators,38 sol-
vents,2 filler loading, functional monomers,19 and methac-
rylate monomers, such as bis-GMA and TEG-DMA.29
Regarding the effect of DC on the resin-dentin bond du-
rability, a moderate correlation was reported between DC
and μTBS reduction of five 1-SEAs and one etch-and-rinse
adhesive after 5000 thermal cycles.6 On the other hand,
increasing the DC by prolonging the light-curing time in-
creased the nanoleakage, but did not increase the durabil-
ity of resin-dentin bonds of 1-SEAs.11 To the extent of the
authors’ knowledge, the influence of the DC of adhesive
resins in 2-SEAs on the durability of bonding when the
same self-etch primer is used has never been investi-
gated. Therefore, the objective of this study was to evalu-
ate the effect of DC on initial and long-term bonding effi-
cacy of 2-SEAs to human dentin, elastic modulus, and
water sorption. The null hypothesis tested was that the DC
of 2-SEAs has no effect on the initial and long-term μTBS
to dentin.
MATERIALS AND METHODS
Two 2-SEAs, Clearfil SE Bond (CSE; Kuraray Noritake; Tokyo
Japan) and Clearfil SE Bond 2 (CSE2; Kuraray Noritake)
were used in this study. Their compositions are shown in
Table 1. CSE and CSE2 both contain camphorquinone (CQ)
as a photo-initiator. However, CSE2 contains an additional
photo-initiator for enhancing the catalyst system and im-
proving the DC of an adhesive resin. The other ingredients
of these adhesives are same.
Measurement of Degree of Conversion (DC)
The polymerization process of the adhesive resins for CSE
or CSE2 was monitored by using a Fourier-transform infra-
red spectrometer equipped with an ATR attachment (Nicolet
6700 FT-IR, Thermo Fisher Scientific; Waltham, MA, USA) at
a resolution of 8 cm-1. The ATR diamond crystal permitted
scanning between 650 and approximately 4000 cm-1. One
drop of the adhesive resins of CSE or CSE2 was placed on
the diamond crystal of the ATR attachment. That is, the
primer was not tested. Photo-activation was performed
using an LED light-curing unit (Pencure 2000, Morita; Tokyo,
Japan) for 10 s. The distance between the light-guide tip
and the top of the diamond crystal was 10 mm (Fig 1).
Time-based spectral acquisition software (Spectrum Time
Base, Perkin-Elmer; Waltham, MA, USA) was used for con-
tinuous and automatic collection of spectra during polymer-
ization at a rate of one spectrum every 0.2 s. Real-time in-
frared spectra were continuously recorded for 40 s after
light activation began. Three separate measurements for
each adhesive were conducted. The light output of the light-
curing unit was 600 mW/cm2 as determined with a radiom-
eter (Demetron LED Radiometer, SDS/Kerr; Orange, CA,
The Journal of Adhesive Dentistry
 Sato et al

Table 1 Chemical composition and application mode of the materials tested

Material Ingredients Procedure

Adhesives Clearfil SE Bond Primer 10-MDP, HEMA, hydrophilic aliphatic dimethacrylates, Apply primer and leave it for
(CSE, Kuraray dl-camphorquinone, water, accelerators, dyes 20 s, then evaporate the water
Noritake; Tokyo with a mild air stream.
Japan)
Clearfil SE Bond 2 Adhesive CSE: 10-MDP, HEMA, bis-GMA, hydrophobic aliphatic Apply adhesive and use a light
(CSE2, Kuraray resin methacrylates, colloidal silica, dl-camphorquinone, air stream to make the adhesive
Noritake) initiators, accelerators film as uniform as possible.
CSE2: 10-MDP, HEMA, bis-GMA, colloidal silica, Light cure adhesive resin for
hydrophobic aliphatic dimethacrylates, dl- 10 s.
camphorquinone, new initiators, accelerators

Composite Clearfil AP-X Bis-GMA, TEG-DMA, camphorquinone,photo-initiators, pigments, Light cure for 40 s.
(Kuraray Noritake) silanated barium glass, silanated silica

10-MDP: 10-methacryloyloxydecyl dihydrogen phosphate; HEMA: 2-hydroxyethyl methacrylate; bis-GMA: bis-phenol A glycidylmethacrylate; TEG-DMA: triethylene
glycol dimethacrylate.

USA). The DC was assessed according to the changes in

absorbance ratio between the peak at 1637 cm-1 (methac- Light tip
rylate C=C stretching) and that at 1608 cm-1 (phenyl C=C
stretching) before and after light curing. The intensities of Adhesive resins
these two bands were integrated based on band height (CSE, CSE2)
methodology, and the change of the band ratio profile with
10 mm Diamond crystal
1637 cm-1/1608 cm-1 was monitored. The degree of con-
version (DC) was calculated by the following equation:

DC (%) = [1– (absorbancecured 1637 cm-1 / absorbancecured

1608 cm-1) / (absorbanceuncured 1637 cm-1 / absorbanceuncured
1608 cm-1)] x 100

Microensile Bond Strength (μTBS) Test

Twenty-four freshly extracted caries-free human molars were
used in this study, according to the protocol approved by
the Human Research Ethics Committee, Tokyo Medical and
Dental University, Japan (No. 725). The occlusal enamel
was removed 90 degrees to the long axis of the tooth by
means of a model trimmer under running water to obtain
midcoronal dentin. The dentin surfaces were prepared with
600-grit SiC paper under running water for 60 s to create a
standardized smear layer. The prepared teeth were then
randomly divided into two groups. The primers of two
2-SEAs (CSE, CSE2) were applied to the dentin surfaces of
each group for 20 s, then the water was evaporated with a
gentle air stream. After that, the bonding agent of CSE and
CSE2 was also applied using a light air stream to make the
adhesive film as uniform as possible, and then irradiated
with a light-curing unit (Pencure 2000, Morita) for 10 s at a
light output > 600 mW/cm2. After light curing the adhesive
resin, a composite, Clearfil AP-X (Shade-A2, Kuraray Nori-
take), was built up incrementally to a height of 5.0 mm to
ensure sufficient bulk for the μTBS test. Each increment
was light cured for 20 s. After storage in 37°C tap water for
24 h, 10 beams (surface area: 0.7 x 0.7 mm) were fabri-
cated from the central part of each bonded tooth using a
low-speed diamond saw (Isomet, Buehler; Lake Bluff, IL,
USA) under water spray coolant. The 10 beams of all
bonded teeth were then randomly divided into three groups.
Infrared beam
Fig 1 Schematic of the polymerized specimens of the adhesive
resins to measure the DC using ATR-FTIR.
All of the beams from each group (3 or 4) were individually
bonded to a tensile testing jig using cyanoacrylate glue
(Zapit, Dental Ventures of America; San Ramon, CA, USA)
mounted in a tabletop testing machine (EZ Test, Shimadzu;
Kyoto, Japan) and subjected to the μTBS test at a cross-
head speed of 1.0 mm/min after water storage at 37°C for
24 h, 6 months, or 1 year. The μTBS was determined by
calculation of the mean of three or four beams from the
same tooth (n = 12). After the μTBS test, all of the frac-
tured specimens were observed using scanning electron
microscopy (SEM; JSM-5310, JEOL; Tokyo, Japan) to char-
acterize their failure modes. The failure modes were classi-
fied as: (1) C-D, cohesive failure in dentin; (2) C-A, cohesive
failure in adhesive resin; (3) C-R, cohesive failure in com-
posite; or (4) I-F, interfacial failure. The area ratios of each
failure mode on the dentin sides were expressed as per-
centages of the overall fractured surface on the SEM micro-
graphs by digitizing and subjecting them to quantitative
image analysis using SemAfore software (Insinöörito-
imisto, J. Rimppi; Ojakkala, Finland).

Vol 19, No 1, 2017 33

Sato et al
Water Sorption Test
Just the adhesive resins of CSE and CSE2 (no primers)
were poured into round silicone molds (8.0 mm in diameter
and 1.5 mm thick) and covered by a thin, transparent Mylar
strip. The adhesive resin in these molds was irradiated for
180 s with an LED light-curing unit (Pencure 2000, Morita)
at a light output > 600 mW/cm2 to fabricated polymerized
resin disks (n = 8). The thickness and diameter of the spec-
imens were measured using a digital caliper (Mitsutoyo;
Kanagawa, Japan), rounded to the nearest 0.01 mm. These
measurements were used to calculate the volume (V) of
each specimen. Water sorption was determined according
to the ISO specification 4049 (2000), except for the speci-
men dimensions.34 Immediately after polymerization and
dry storage for 24 h in a container filled with anhydrous sil-
ica gel, the disks were repeatedly weighed at intervals of
24 h until a constant mass (M1) was obtained.
After measuring the initial dry mass (M1), the disks were
individually immersed in tap water at 37°C for 24 h. After
water storage for 24 h, the disks were gently wiped with
absorbent paper, weighed, and put back in tap water. This
regime was repeated and continued until a constant wet
mass (M2) was obtained. Water sorption (WS) was calcu-
lated using the following formula:
WS = (M2 − M1)/V
where M1 is the initial dry constant mass (μg) before water
immersion, M2 is the constant wet mass (μg) after water
immersion, and V is the volume of the specimen in mm3.
Water sorption values were expressed in g/mm3.
Three-point Flexural Test
The adhesive resins of CSE and CSE2 without mixing with
primers were poured into silicone molds (5 x 18 x 2 mm)
and covered by a transparent thin Mylar strip. The adhesive
resin in these molds was irradiated for 180 s with an LED
light-curing unit (Pencure 2000, Morita) with a light out-
put > 600 mW/cm2 to fabricate sixteen polymerized resin
beams. Immediately after polymerization and dry storage in
a container filled with anhydrous silica gel for 24 h, the six-
teen specimens from each adhesive resin were divided into
two groups. Half of specimens (dry group) were subjected to
a three-point flexural test for measuring modulus of elastic-
ity (E) (n = 8). The rest of the specimens (wet group) were
subjected to a three-point flexural test after storage in tap
water for 1 month, as 1 month is necessary in the water
sorption test for the weight of specimens to reach a pla-
teau. The three-point bending test was performed by cen-
trally loading the beams using a universal testing machine
(Autograph AGS-H, Shimadzu) at a displacement rate of
1 mm/min, sufficient to induce a 3% strain. The compres-
sive force necessary to induce a 3% strain in the beam was
measured. Load displacement values were converted to
stress and strain. The modulus of elasticity values was cal-
culated as the slope of the linear portion of the stress-
strain curve from the following formula:
E = FL3/4Dbh3
34
where F is the force (N), L is the span length (10 mm), b is the
width of the plate (5.0 mm ± 0.1 mm), D is the vertical deflec-
tion (mm) of the plates, and h is the thickness (2.0 mm). The
modulus of elasticity was expressed in MPa. The strain (ε)
produced by three-point bending was calculated as:
ε = 6hd/L
where h is the thickness of the plate (mm), d is the dis-
placement of the plate (mm), L is the span length of the
plate between the supports (5.0 mm).
Differences in the moduli of elasticity of the specimens
with and without immersion of specimens in water were
calculated as percent decrease of the modulus of elasticity
(%ΔE), as follows:
ZE= - {(Ewet-Edry)/Edry} x 100
where Ewet was the modulus of elasticity of wet-group spec-
imens and Edry was the modulus of elasticity of dry-group
specimens.
Statistical Analysis
The maximum DC and water sorption data were analyzed
using Student’s t-test (α = 0.05). Normal distributions of
elastic modulus and μTBS were indicated by the Shapiro-
Wilk test (p > 0.05). Elastic modulus was analyzed using
two-way ANOVA (α = 0.05), and the μTBS values were sta-
tistically analyzed using two-way ANOVA and the t-test with
Bonferroni correction for multiple comparisons to test the
effect of the adhesives and the periods of water storage
(24 h, 6 months, 1 year) (α = 0.05).
RESULTS
Figure 2 shows the change of DC of the two test adhesive
resins over 40 s after the light activation began. Student’s
t-test revealed that the maximum DC of the adhesive resin
of CSE2 was significantly higher than that of CSE
(p < 0.001) (Table 2). Table 2 shows the mean μTBS of the
two 2-SEAs to dentin as a function of the water-storage pe-
riod. There were no pre-test failures during specimen prep-
aration. Two-way ANOVA revealed that μTBS was influenced
by the adhesives (p < 0.001) and water storage periods
(p < 0.001). In addition, there was a significant interaction
between the adhesives and water storage periods
(p < 0.001). CSE2 showed significantly higher μTBS than
CSE after each water storage period of 24 h, 6 months, and
1 year (p < 0.001). In CSE specimens, the μTBS after water
storage for 1 year was significantly lower than the μTBS
after water storage for 24 h (p < 0.001). However, there
were no significant differences between the μTBS after stor-
age for 24 h, 6 months, and 1 year (p = 0.25, 0.94, and
1.00, respectively) in CSE2 specimens. The failure mode
percentages are summarized in Fig 3. C-A failures were only
seen in CSE specimens and increased over time, while C-A
failures were never seen in CSE2 specimens at any time
period. CSE2 specimens show similar failure mode distribu-
tion among all the time periods. The water sorption values
The Journal of Adhesive Dentistry
 Sato et al

Failure mode
80
10 s light irradiation CSE 24h
70
Degree of conversion (%)

60 CSE 6m
50
CSE 1y C-D
40
C-A
30 C-R
CSE2 24h
I-F
20 CSE2 CSE

10 CSE2 6m

0
0 5 10 15 20 25 30 35 40 CSE2 1y

Time (s) 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100%

Fig 2 The DC (%) vs time (s). Horizontal bar shows light-curing du- Fig 3 Percentage of failure modes of two 2-SEAs. C-D: cohesive
ration in s. CSE: Clearfil SE Bond; CSE2: Clearfil SE Bond 2. failure in dentin, C-A: cohesive failure in adhesive; C-R: cohesive fail-
ure in composite; I-F: interfacial failure. m: months; y: year.

Table 2 The degree of conversion (DC) and microtensile bond strengths (μTBS) of two 2-SEAs tested in this study

Materials DC (%), n = 3 μTBS (MPa), n = 12

24 h 6 months 1 year
CSE 56.0 ± 0.1A 71.5 ± 7.2Aa 66.6 ± 2.6Aab 53.1 ± 4.2Ab

CSE2 66.8 ± 0.2B 80.6 ± 5.1Ba 78.7 ± 5.5Ba 76.2 ± 4.7Ba

Values are mean ± SD. Different superscript letters indicate statistically significant differences in rows (lowercase) and columns (capital) (p < 0.05). CSE:
Clearfil SE Bond; CSE2: Clearfil SE Bond 2.  

Table 3 Water sorption, modulus of elasticity, and reduction rate of elastic modulus of two 2-SEAs tested in this
study

Materials Water sorption Modulus of elasticity (MPa), n = 8 Reduction of modulus

(μg/mm3) (n=8) of elasticity (%)
Dry Wet
CSE 72.8 ± 1.7A 1345 ± 93Aa 1058 ± 98Ab 21.4

CSE2 65.0 ± 3.1B 1568 ± 191Ba 1330 ± 212Bb 15.2

Values are mean ± SD. Different superscript letters in each column (capital) or row (lowercase) indicate statistically significant differences among storage periods
(p < 0.05). Reduction of modulus of elasticity (%): [{modulus of elasticity (wet group)-modulus of elasticity (dry group)}/modulus of elasticity (dry group)] x 100.
CSE: Clearfil SE Bond; CSE2: Clearfil SE Bond 2.

and moduli of elasticity (dry vs wet) of the polymerized ad-
hesive resin of the two 2-SEAs are shown in Table 3. Stu-
dent’s t-test revealed that water sorption of CSE2 was sig-
nificantly lower than that of CSE (p < 0.05). Two-way ANOVA
showed that modulus of elasticity was influenced by adhe-
sive (p < 0.001) and storage condition (p < 0.001). In addi-
tion, there was no significant interaction between the adhe-
sives and storage conditions (p = 0.67). CSE2 showed a
significantly higher modulus of elasticity than CSE at each
storage condition (p < 0.001). The elastic modulus of both
adhesive resins was reduced by water storage (p < 0.001).
The percent reduction in modulus of elasticity was 21.4%
(CSE) and 15.2% (CSE2).
DISCUSSION
In this study, the DC of CSE2 was significantly higher than
CSE. Regarding bond strengths, there was a significant in-
teraction (materials x water storage time) (p < 0.001), and
there were no significant differences between 24 h,
6 months, and 1 year (p = 0.25, 0.94, and 1.00, respect-

Vol 19, No 1, 2017 35

Sato et al
ively) in CSE2 specimens, while in CSE specimens, the
μTBS after water storage for 1 year was significantly lower
than the μTBS after water storage for 24 h (p < 0.001).
Therefore, the null hypothesis that DC of 2-SEAs has no
effect on the initial and long-term μTBS of 2-SEAs to dentin
is rejected. In the literature, some studies found a correla-
tion between DC and the bond strengths of both self-etch
adhesives and etch-and-rinse adhesives,11 but another
study failed to find any correlation.25 This indicates that
the different compositions of various adhesives have a
major impact on these results, because bond strength is
multifactorial.
From the results of the DC measurements, it was found
that that the DC of CSE2 was significantly higher than that
of CSE (p < 0.001). In this study, an LED light-curing unit
was used because of the increasing use of LED lights by
clinicians. The DC of CSE significantly decreased when ir-
radiation was carried out using three different LED curing
lights instead of a quartz-tungsten-halogen curing light
(QTH), because of its narrower spectrum of wavelengths
which include the absorption range of the photo-initiators
used in the adhesive (QTH: 74.1%; LEDs: ca 64.7%-
65.5%).7 The DC of CSE (56.0%) evaluated in this study is
lower, but this would be due to the different light irradiation
duration. Both adhesive resins of CSE and CSE2 contain
CQ, which is the most widely and successfully used photo-
initiator in dental restorative resins.16 Moreover, a newly
developed photo-initiator has been added to CSE2. The sig-
nificantly higher bond strengths at any water storage period
and the DC of CSE2 compared to CSE may be due to the
additional photo-initiator of the adhesive resin of CSE2. On
the other hand, the residual component of the primer on
the dentin surface applied prior to the application of adhe-
sive resin may affect the DC of the adhesive resins, and
further research is needed.26
The adhesives containing 10-methacryloyloxydecyl dihy-
drogen phosphate (10-MDP) may chemically bind to the hy-
droxyapatite of dental hard tissues, providing higher me-
chanical strength37 and bonding stability40 due to the
presence of nanolayering of 10-MDP-calcium salts in the
resin-dentin interface. However, some researchers reported
that the presence of nanolayering is unlikely to contribute
to the overall resin-dentin durability.35 In fact, some in vitro
studies have shown that the μTBS of CSE, which contains
10-MDP, to dentin decreases after long-term water stor-
age.3 The μTBS of CSE2 after both 24 h and 1 year was
significantly higher than that of CSE. Furthermore, there
was a significant interaction (materials x water storage
time) (p < 0.001), which revealed that the resin-dentin bond
created by CSE2 had superior durability compared to CSE.
We speculate that – compared to CSE9,17 – a well-polymer-
ized, stiffer adhesive layer and hybrid layer created by CSE2
should lead to better load transfer across the bonded inter-
face over time, and resist higher tensile stresses under
load, due to its higher mechanical properties. For different
commercially available 2-SEAs, a significant correlation was
found between the mechanical properties of adhesive res-
ins and their resulting μTBS.33 Polymerization of the resin
36
monomers both within the adhesive layer and in the under-
lying demineralized dentin would play an important role of
the resin-dentin bond, since previous studies have also re-
ported that the DC of adhesive resins and the hybrid layer
are correlated with the resin-dentin μTBS.11 It is speculated
that a higher DC in adhesive resins may improve the quali-
ties of not only the adhesive layer but also hybrid layer.
The water sorption of CSE2 (72.8 ± 1.7 μg/mm3) was
found to be significantly lower than that of CSE
(85.8 ± 9.9 μg/mm3) (p = 0.03, Table 3). Moreover, the re-
sults of the three-point bending test showed that the elastic
modulus of CSE2 was significantly higher than that of CSE
before and after water storage (p < 0.001). These findings are
in accordance with previous studies showing that higher DCs
of adhesive resins are correlated to lower water sorption21
and better mechanical properties.9,10 We speculate that
CSE2 creates more cross linking of the polymerized adhesive
resins, resulting in less water sorption and higher elastic
modulus compared to CSE. The reason why the water-stor-
age–induced percentage reduction in elastic modulus for
CSE2 (15.2%) was lower than that of CSE (21.4%) may also
be because of the greater hydrophobicity of CSE2, since the
percentage reduction in the mechanical properties of adhe-
sives are related to their water sorption values.13 Interest-
ingly, the elastic modulus of CSE2 even after water storage
was higher than that of CSE before water storage, which
means that the polymerized adhesive resin of CSE2 with a
higher DC leads to higher mechanical properties than in CSE,
with its lower DC regardless of moisture conditions, and can
contribute to the mechanical stability of the resin-dentin bond.
Failure mode observations using SEM provided important
information regarding which layer of the bonded assembly is
the major site of crack initiation and propagation. Cohesive
failures within the adhesive layer were only seen in the CSE
specimens, while it was not seen in the CSE2 specimens.
This finding may also result from the additional photo-initia-
tor in CSE2.
CONCLUSION
The higher DC of adhesive resins of 2-SEA resists water
aging and improves the initial bond strengths and durability
of resin-dentin bonding. This may indicate the potential that
manufacturers can reach with 2-SEAs, if they use neat adhe-
sives as a sealer for 2-SEAs and improve their hydrophilic/
hydrophobic photo-initiator/accelerators. As a previous study
found that the bond strength data after 5-year in vitro aging
are more clinically relevant and may predict the long-term
clinical performance,36 the 5-year durability of resin-dentin
bonds of CSE2 should be explored in future studies.
ACKNOWLEDGMENTS
This work was supported by Grant-in-Aid for Scientific Research (C)
#26462873 and Grant-in-Aid for Scientific Research (C) #15K11108
from the Ministry of Education, Culture, Sports, Science and Technol-
ogy of Japan.
The Journal of Adhesive Dentistry

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Clinical relevance: The higher DC of Clearfil SE Bond 2
may facilitate better clinical performance.
37