Effects of Selective Phosphoric Acid Etching on Enamel

Using “No-wait” Self-etching Adhesives

Takaaki Satoa / Tomohiro Takagakib / Masaomi Ikedac / Toru Nikaidod / Michael F. Burrowe /
Junji Tagamif

Purpose: To evaluate the effect of selective phosphoric acid etching on enamel using “no-wait” self-etching adhesives.
Materials and Methods: Clearfil Universal Bond Quick (UBQ, Kuraray Noritake) or G-Premio BOND (GPB, GC) was
applied to ground human enamel surfaces. The adhesives were used in 3 modes: no-waiting self-etching mode
(UBQ-0 or GPB-0), 10-s self-etching mode (UBQ-10 or GPB-10), and the selective-etch mode with phosphoric acid
etching (UBQ-PA or GPB-PA). After an acid-base challenge, the morphological attributes of the interface were exam-
ined using scanning electron microscopy (SEM) to characterize the acid-base resistant zone (ABRZ). Microshear
bond strength (μSBS) testing to enamel and ultimate tensile strength (UTS) of the adhesive resin were carried out.
Results: ABRZ formation was confirmed in all groups. UBQ-PA and GPB-PA created thicker ABRZ with no funnel-
shaped erosion beneath the adhesive-enamel interface. μSBS of UBQ-PA was significantly higher than UBQ-0 and
UBQ-10 (p < 0.05). However, there were no statistically signficant differences in μSBS among GPB-0, GPB-10, and
GPB-PA. For the UTS, UBQ was significantly higher than GPB.
Conclusion: Selective phosphoric acid etching created a stable adhesive-enamel interface. The no-waiting self-etch-
ing concept adhesive led to a thinner ABRZ. The results of μSBS suggest that phosphoric acid etching effects on
enamel are material dependent.
Keywords: enamel, multimode, no-waiting self-etching concept, phosphoric acid, acid-base resistant zone, micro-
shear bond strength test.

J Adhes Dent 2018; 20: 407–415. Submitted for publication: 05.04.18; accepted for publication: 17.08.18
doi: 10.3290/j.jad.a41359

a Medical Staff Dentist, Cariology and Operative Dentistry, Graduate School of

Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo,
Japan. Participated in experimental design, performed the experiments, wrote
the manuscript.
b Assistant Professor, Cariology and Operative Dentistry, Graduate School of
Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo,
Japan. Idea, hypothesis, experimental design, proofread the manuscript.
c Junior Assistant Professor, Oral Prosthetic Engineering, Graduate School, Fac-
ulty of Dentistry, Tokyo Medical and Dental University, Tokyo, Japan. Per-
formed statistical analysis, contributed substantially to discussion.
d Lecturer, Cariology and Operative Dentistry Department, Graduate School of
Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo,
Japan. Idea, hypothesis, experimental design, proofread the manuscript.
e Clinical Professor in Prosthodontics, Faculty of Dentistry, The University of
Hong Kong, Hong Kong SAR, China. Contributed substantially to discussion,
proofread the manuscript.
f Professor, Cariology and Operative Dentistry Department, Graduate School
of Medical and Dental Sciences, Tokyo Medical and Dental University,
Tokyo, Japan. Supported the experiments, contributed substantially to dis-
cussion.
Correspondence: Tomohiro Takagaki, Cardiology and Operative Dentistry,
Graduate School of Medical and Dental Sciences, Tokyo Medical and
Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan.
Tel: +81-3-5803-5483. e-mail: takagaki.ope@tmd.ac.jp
S elf-etching adhesives (SEAs) are widely accepted, and
have been classified into two types: 2-step self-etching
adhesives (2-SEAs) and one-bottle self-etching adhesives
(1-SEAs). Recently, the multimode concept has been intro-
duced, where some 1-SEAs can be used either in self-etch-
ing mode, selective-etch mode, or etch-and-rinse
mode.3,18,24 This multimode capability enables the clinician
to apply the adhesive using the selective enamel etching
technique, which includes the advantages of the etch-and-
rinse technique on enamel.16
Recently, some 1-SEAs have been employed with the op-
tion of using the self-etching technique for bonding without
waiting, or after leaving the adhesive undisturbed for
10 s.27 Huang et al12 evaluated no-waiting self-etching ad-
hesives to dentin using the two alternative self-etching
modes (no-waiting or 10-s self-etch). They reported that suf-
ficient bond strength to dentin may be achieved in the no-
waiting self-etching mode. However, the no-waiting self-etch-
ing mode may not represent the best use of the
adhesive.12 On the other hand, few studies have focused
on enamel with no-waiting self-etching adhesives.

Vol 20, No 5, 2018 407

Sato et al
Table 1 Composition of the adhesives used in the study
Material pH Manufacturer Composition
CLEARFIL Universal 2.3 Kuraray, 10-MDP, bis-GMA, HEMA, hydrophilic amide monomer, colloidal silica,
Bond Quick Noritake Dental ethanol, dl-camphorquinone, initiators, accelerators, water, sodium fluoride
G-Premio Bond 1.5 GC 10-MDP, 4-MET, methacrylate acid ester, distilled water,
10-methacryoyloxydecyl dihydrogen thiophosphate, acetone, photoinitiators,
silica
Bis-GMA: bisphenol A diglycidylmethacrylate; HEMA: hydroxyethyl methacrylate; 10-MDP: 10-methacryloyloxydecyl dihydrogen phosphate; 4-MET: (4-[2-(methac-
ryloyloxy)ethoxycarbonyl]phthalic acid).
1-SEAs are regarded as ultra-mild self-etching adhe-
sives;19 however, the bonding mechanism to enamel relies
on micromechanical interlocking provided by the resin tags
in acid-etched enamel. Furthermore, some studies con-
cluded that phosphoric acid etching prior to the application
of self-etching adhesives (selective phosphoric acid etching)
improves enamel bond strength.3,8,32 The mild etching abil-
ity has raised some concerns with regard to the ability of
these 1-SEAs to form a stable bond and marginal sealing of
the restoration without selective phosphoric acid etching.35
Tsuchiya et al34 first observed ultramorphological
changes at the interface between adhesive and dentin after
an acid-base challenge by using scanning electron micros-
copy (SEM). They found a new zone resistant to acid-base
attack, the so-called acid-base resistant zone (ABRZ) adja-
cent to the dentin hybrid layer in self-etching adhesives. The
ABRZ was thought to play an important role in the preven-
tion of recurrent caries, sealing of restoration margins, and
promotion of restoration durability.21,22 Li et al15 found that
a similar zone, “enamel ABRZ”, formed at the adhesive-
enamel interface with 2-SEAs containing 10-methacryloyl-
oxydecyl dihydrogen phosphate (10-MDP). It has been re-
ported that 2-SEAs containing 10-MDP created a stable
enamel ABRZ, whereas the 1-SEAs containing 10-MDP cre-
ated thinner ABRZs with funnel-shaped erosion, indicating a
weak area vulnerable to acid-base attack beneath the adhe-
sive-enamel interface. However, this erosion did not occur
in 1-SEAs with selective phosphoric acid etching.28
The purpose of this study was to evaluate the effect of
selective phosphoric acid etching on enamel using no-wait-
ing self-etching adhesives. The null hypotheses of this study
were that 1) the extension of application time improves the
adhesive-enamel interface, 2) selective phosphoric acid
etching has no effect on the adhesive-enamel interface
when the no-waiting self-etching adhesive is used, and 3) all
no-waiting self-etching adhesives show the same trend.
MATERIALS AND METHODS
Materials
Non-carious human third molars and premolars were col-
lected after obtaining the individuals’ informed consent ac-
cording to a protocol approved by the Institutional Review
408
Lot No.
T160128
1603091
Board of Tokyo Medical and Dental University (D2013-022).
The teeth were stored at 4°C in 0.01% thymol solution, and
used within 1 month following extraction.
Two kinds of 1-SEAs were used in this study (Table 1),
Clearfil Universal Bond Quick (UBQ, Kuraray Noritake; Tokyo,
Japan) and G-Premio BOND (GPB, GC; Tokyo, Japan). Both
adhesives can be applied according to the no-waiting self-
etching concept. UBQ contains 10-MDP, while GPB contains
10-MDP and (4-[2-(methacryloyloxy)ethoxycarbonyl]phthalic
acid) (4-MET) as functional monomers. For GPB, no-waiting
self-etching mode is recommended in the Japanese version
of the manufacturer’s instruction, and 10-s self-etching
mode is recommended in the international version.
Specimens were randomly divided into six groups. For
the no-waiting self-etching mode, the adhesive was applied
for less than 1 s and immediately air dried with no time
lapse between steps (UBQ-0 or GPB-0). For the 10-s self-
etching mode, the adhesive was applied on the enamel sur-
face for 10 s (UBQ-10 or GPB-10). For the selective-etch
mode, 37% phosphoric acid (K-etchant Gel, Kuraray Nori-
take; Tokyo, Japan) was applied for 10 s, then rinsed and
dried. Each adhesive was applied and air dried without wait-
ing (UBQ-PA or GPB-PA). An LED light-curing unit (Valo LED
Curing Light, Ultradent; South Jordan, UT, USA) was used in
high power mode (1400 mW/cm2) to light cure the adhe-
sive for 10 s.
Microshear Bond Strength (μSBS) Test
Sample preparation was carried out as shown in Fig 1. A
2-mm-thick cuspal enamel slice was sectioned using a low-
speed diamond saw (Isomet, Buehler; Lake Bluff, IL, USA).
In order to standardize the smear layer, the enamel sur-
faces were wet polished with 600-grit SiC paper for 1 min.
The slices were randomly divided into six groups. Prior to
polymerization of the adhesive resin, a tygon tube (R-3603,
Professional Plastics; Fullerton, CA, USA) with an internal
diameter of 0.79 mm and a height of 0.5 mm was placed
on each bonded area. After the adhesive was light cured, a
hybrid resin composite, Clearfil AP-X (shade A2, Kuraray
Noritake), was inserted into the tube and polymerized for
40 s. After light curing, the tygon tube was removed, and
the composite rods checked for defects. All bonded speci-
mens were stored in 37°C distilled water for 24 h before
testing. The specimens were attached to the testing device
The Journal of Adhesive Dentistry
 Sato et al

Microshear bond strength test

24 h storage in
Cut
water 37°C
SEM

#600 SiC paper Bonding,

composite buildup, Failure mode
light curing observation

Ultimate tensile strength test

Air blowing, Universal testing machine; EZ-SX

light curing Crosshead speed of 1.0 mm/min

24 h storage
with silica gel

Silicone mold Beam specimen

Width: 1 mm
Height: 1 mm
Length: 10 mm

Fig 1 Sample preparation for microshear bond strength testing and ultimate tensile strength testing.

(EZ-SX, Shimadzu; Kyoto, Japan) and subjected to micro-
shear bond testing at a crosshead speed of 1 mm/min, as
previously described.23,29
The mean bond strengths were calculated, then statisti-
cally analyzed in PASW Statistics version 18 (IBM; Armonk,
NY, USA) using two-way ANOVA and t-tests with Bonferroni
correction at a significance level of α = 0.05 (n = 15).
The failure mode was determined using SEM (JSM-
IT100, JEOL; Tokyo, Japan). Failure modes were divided into
four categories: bond-enamel interface (B/E); resin-compos-
ite-bond interface (R/B); cohesive failure in bond (B); cohe-
sive failure in enamel (E). The area of each failure type (%)
was calculated and statistically analyzed using Dunn’s test
with Bonferroni correction at a significance level of
α = 0.05.
Ultimate Tensile Strength (UTS) Test
GPB and UBQ were poured into beam-shaped silicone
molds (Fusion II, putty type, GC) with dimensions of 10 mm
length × 1 mm width × 1 mm height (Fig 1). The solvents of
each adhesive were evaporated with a 3-way dental air sy-
ringe for 30 s held at a distance of 15 cm with an air pres-
sure of 3.8 kgf/cm2. After covering the top of the surface
with thin transparent strips (Hawe Striproll, Transparent
Striproll No.687, Kerr; Orange, CA, USA) and a glass slab,
the light guide was placed directly on the surface and light
cured for 1.5 min from each side.11,17 Twelve specimens
were manufactured for each material. After polymerization,
all specimens were kept in a desiccator with silica gel for
24 h, then subjected to the UTS test using a universal test-
ing machine at a crosshead speed of 1.0 mm/min until fail-
ure. The UTS of the adhesives was calculated as:
UTS = F/A, where F is maximum stress (N) and A is the
cross-sectional area of the specimen (mm2). As for μSBS,
the UTS was expressed in MPa. Statistical analysis was
carried out using the t-test (α = 0.05).
SEM Observation of ABRZ
Sample preparation is illustrated in Fig 2. Roots were re-
moved and crowns were sectioned mesiodistally using an
Isomet low-speed saw (Buehler). In order to prepare the
mid-coronal buccal or lingual enamel surfaces for bonding,
the specimens were embedded in epoxy resin (EpoxiCure,
Buehler) with buccal or lingual surfaces facing out, and then
ground with wet 600-grit SiC paper until flat enamel sur-
faces were exposed. The specimens were randomly divided
into six groups. An LED light-curing unit was used to poly-
merize the bonding agent after application. Clearfil Majesty
LV (shade A2, Kuraray Noritake) was applied on top of the
cured adhesive (approximately 2 mm thick) and cured for
20 s. After storage in 37°C distilled water for 24 h, each
specimen was sectioned perpendicular to the bonding inter-
face into two halves and embedded in the epoxy resin. The
specimens were then polished with SiC papers from 600- to
1200-grit, then subjected to an acid-base challenge to ex-
amine the acid-base resistance of the interfacial structure.
For the acid challenge, each specimen was stored in buff-
ered demineralizing solution (pH 4.5, 2.2 mmol/l CaCl2,
2.2 mmol/l NaH2PO4, and 50 mmol/l acetic acid) for 4.5 h.
For the base challenge, the specimens were immersed in

Vol 20, No 5, 2018 409

Sato et al
SEM observation of ABRZ
Cut Embed
Acid-base challenge Polishing
Diamond paste
(1/4 μm)
Demineralization solution (pH 4.5), 4.5 h
6% NaOCl, 20 min with ultrasonication
Fig 2 Sample preparation for SEM observation of ABRZ on enamel.
6% NaOCl for 20 min with ultrasonic agitation to remove
any debris on the demineralized structures or enamel pro-
teins, and finally rinsed with running water for 30 s. Follow-
ing this, a 4-META/MMA-TBB resin (Super Bond C&B, Sun
Medical; Moriyama, Japan) was applied without acid etching
the surface, in order to prevent wear or fracture of the re-
maining structure during polishing. The specimens were
then cut perpendicular to the adhesive-enamel interface,
and polished with diamond pastes (Struers; Ballerup, Den-
mark) down to 0.25-μm particle size. Argon-ion etching (EIS-
200ER, Elionix; Tokyo, Japan) was used for the polished
surfaces to bring the adhesive-enamel interface into sharp
relief, with an accelerating voltage of 1 kV and an ion cur-
rent density of 1.5 mA/cm2 for 30 s. The specimens were
finally gold sputter-coated, and the morphological structure
of the interface after the acid-base challenge was observed
using SEM with an accelerating voltage of 20 kV.
SEM Observation of the Enamel Surface after
Application of Adhesive
The enamel surfaces of six premolars were wet polished
with 600-grit SiC paper until flat enamel surfaces were ex-
posed, then treated with each bonding regimen as de-
scribed above. Without light curing, the surface was ultra-
sonically rinsed with absolute acetone for 1 min in order to
dissolve the adhesive and dehydrate the specimens for
SEM observation. After air drying for 10 s, samples were
sputter-coated with gold and examined using SEM to ob-
serve the surface structure and etching pattern.
410
Bonding, composite buildup,
light curing Cut and embed
24 h storage
in water 37°C
#600 SiC paper #1200 SiC paper
SEM; JSM-IT100
Ar-ion etching
(EIS-220Et, Elionix)
RESULTS
μSBS Test
The μSBS and failure modes are shown in Tables 2 and 3.
UBQ-PA provided significantly higher μSBS than did UBQ-0
(p < 0.001) and UBQ-10 (p = 0.008). However, there were
no statistically significant differences between GPB-0, GPB-
10, and GPB-PA (p > 0.05). There was a significant differ-
ence between UBQ-PA and GPB-PA (p = 0.004).
The fracture at the bond-enamel interface (B/E) was the
most frequent failure mode in all groups, with more than
70% of total bonding area. Then, GPB bonded under selec-
tive-etch mode showed cohesive failure in the bond (B) of
approximately 1/4 of the total bonding area, whereas <14%
of such fractures was detected in others (p = 0.080). In
contrast, 12.7% of the total bonding area in UBQ-PA failed
cohesively in enamel, which was statistically significantly
different from GPB-PA (p = 0.010).
UTS Test
The UTS values are shown in Table 4. The t-test showed
that UBQ had significantly higher UTS than did GPB
(p < 0.001).
SEM Observation of ABRZ
The interfacial morphologies of each group after acid-base
challenge are shown in Fig 3. ABRZ and an outer lesion
(OL), created by mineral loss due to the acid-base chal-
lenge, were observed in all groups. The depth of the OL
ranged from 15 to 20 μm in all groups. ABRZ in UBQ-PA
The Journal of Adhesive Dentistry
 Sato et al

Table 2 Means and standard deviations of microshear bond strength (MPa)

UBQ-0 UBQ-10 UBQ-PA GPB-0 GPB-10 GPB-PA

21.4 ± 7.2a 23.9 ± 8.1b 32.0 ± 8.2a,b,c 24.7 ± 7.5 27.7 ± 7.7 24.4 ± 7.5c

Same superscript letters indicate significant differences (two-way ANOVA and t-tests with Bonferroni correction, α = 0.05).

Table 3 Failure mode (area %) Table 4 Mean values of ultimate tensile strength (MPa)

UBQ-0 UBQ-10 UBQ-PA GPB-0 GPB-10 GPB-PA UBQ GPB

B/E 84.7 79.3 72.7 86.0 84.7 70.7 22.8 ± 4.3a 8.8 ± 2.7a

R/B 4.7 6.7 2.6 0.0 1.3 2.0 Same superscript letters indicate significant differences (t-test, α = 0.05).

B 9.3 14.0 12.0 13.3 14.0 27.3

E 1.3 0.0a 12.7a,b,c 0.7 0.0b 0.0c

B/E: bond-enamel interface; R/B: resin-composite/bond interface; B: co-

hesive failure in bond; E: cohesive failure in enamel. Same superscript
letters indicate significant differences (Dunn’s test with Bonferroni correc-
tion, α = 0.05)

Fig 3 Ultrastructure of adhesive-enamel interface after acid-base challenge (3500X magnification). a: UBQ-0; b: UBQ-10; c: UBQ-PA; d: GPB-
0; e: GPB-10; f: GPB-PA. OL and ABRZ (between white triangles) were observed in all groups. ABRZ in c and f were thicker than a, b, c, and d.
The funnel-shaped erosion (white arrows) was detected in a, b, c, and d. The top area of erosion was wider than bottom area in d and e.
B: bonding, CR: composite resin, D: dentin, OL: outer lesion.

and GPB-PA (Figs 3c and 3f) were thicker than in the other
groups. In UBQ-0, UBQ-10, GPB-0 and GPB-10, funnel-
shaped erosion or defects along the interface were de-
tected beyond the OL, where enamel was dissolved and
detached from the bonding layer. In contrast to UBQ-0 and
UBQ-10 (Figs 3a and 3b), the top area of erosion was
wider than the bottom area in GPB-0 and GPB-10 (Figs 3d
and 3e). On the other hand, no gap or defect was formed
beneath bonding interface in UBQ-PA and GPB-PA (Figs 3c
and 3f).
SEM Observation of the Enamel Surface after
Application of Adhesive
Figures 4 and 5 show the enamel surface after treatment.
Where phosphoric acid etching was not used, a smear layer
covered the entire surface, and scratches from SiC paper

Vol 20, No 5, 2018 411

Sato et al

a b c

d e f

Fig 4 SEM observation of the enamel surface after application of adhesive (5000X magnification). a: UBQ-0; b: UBQ-10; c: UBQ-PA; d: GPB-
0; e: GPB-10; f: GPB-PA. The entire surface was covered by a smear layer, and polishing scratches are clearly visible in UBQ-0 and UBQ-10.
GPB-0 shows a shallow, non-uniform etching pattern. The honeycomb pattern is evident on surfaces of UBQ-PA and GPB-PA.

a b c

d e f

Fig 5 SEM observation of the enamel surface after application of adhesive (10,000X magnification). a: UBQ-0; b: UBQ-10; c: UBQ-PA; d:
GPB-0; e: GPB-10; f: GPB-PA. Enamel crystallites were identified on the surface. The entire surface was covered by a smear layer in UBQ-0,
UBQ-10 and GPB-0. For GPB-10, exposed crystallites within enamel prisms were observed, but a honeycomb pattern was not apparent. In
UBQ-PA and GPB-PA, spatial repetition of regular patterns created a multitude of deep pits, forming a honeycomb structure.

were observed regardless of application time (Figs 4a, 4b, solution of enamel prisms (Figs 4c and 4f). For GPB-10
4d, and 4e). Surfaces etched with phosphoric acid clearly (Fig 5e), exposed crystallites within enamel prisms were
exhibited the honeycomb pattern caused by preferential dis- observed, but a honeycomb pattern was not apparent.

412 The Journal of Adhesive Dentistry


DISCUSSION
Enamel is the hardest tissue in the human body, being
composed of 92–96% inorganic matter or mineral phase,
and 4% of organic material and plasma by weight. The min-
eral phase consists primarily of calcium phosphate in the
form of hydroxyapatite crystals, which are carbonated or
fluoridated.10 The enamel microstructure consists of crys-
tals arranged in prisms or rods which run approximately per-
pendicular to the dentin-enamel junction.4 The interfacial
area between prisms is protein rich, and termed interpris-
matic enamel.7 In the prismless enamel layer, the hydroxy-
apatite crystals are parallel to each other and perpendicular
to the enamel surface.4 Self-etching adhesives are less
acidic than etch-and-rinse adhesives that use phosphoric
acid etching. To improve the bonding performance to
enamel, selective enamel etching has been regarded as a
strategy for optimizing enamel bond strength of mild univer-
sal adhesives. On the other hand, when phosphoric acid
etching was applied to the parallel prismatic enamel, the
enamel surface appeared to be over-etched, resulting in
reduced bond strengths.30 The depth of enamel demineral-
ization might be influenced by the orientation of enamel
prisms.5 In order to avoid confusion, crosscut enamel sur-
faces were used in this study.
The no-waiting self-etching adhesive provides the clin-
ician with the alternatives of using the self-etching tech-
nique for bonding to dentin without waiting for the adhe-
sive to interact with the bonding substrate, or after leaving
the adhesive undisturbed for 10 s.27 Since self-etching
adhesives contain acidic functional monomers and the pH
of these solutions is low, the thickness of the demineral-
ized layer might be affected by the application time. An
increase in adhesive application time significantly influ-
ences the enamel bond strength of 1-SEA.33 SEM observa-
tion of the enamel surface revealed that scratches still
remain on the enamel surface without phosphoric acid
treatment. The surface of UBQ-0 showed little difference
to UBQ-10 (Figs 5a and 5b). In contrast, exposed crystal-
lites within enamel prisms were observed in GPB-10
(Fig 5e). The results of the μSBS test showed that there
were no significant differences between UBQ-0 and UBQ-
10, or GPB-0 and GPB-10. However, in both UBQ and GPB,
μSBS of specimens treated with 10-s application tended
to be higher than that of the no-waiting self-etching tech-
nique. A recent study reported that although reducing ap-
plication time increases user friendliness, using the con-
ventional 10-s application time improved bonding
performance when compared with the no-waiting self-etch-
ing technique.12 The pH of GPB is 1.5. This low pH may
contribute to exposing crystallites in GPB-10, which might
improve the bonding performance due to mechanical inter-
locking. Meanwhile, little difference was observed be-
tween UBQ-0 and UBQ-10. UBQ contains a new hydrophilic
amide monomer, which may penetrate easily into moist
tooth substrates. Therefore, monomer penetration may be
enhanced by 10-s application, yielding a chemical reaction
with the tooth substrate.
Vol 20, No 5, 2018
Sato et al
UBQ-PA showed significantly higher μSBS than did UBQ-0
or UBQ-10. A previous study reported that selective phos-
phoric acid etching improved the bond strength of universal
adhesives.26 SEM observations of the enamel surface re-
vealed that phosphoric acid etching treatment created the
honeycomb pattern on enamel surface. These phenomena
demonstrate that phosphoric acid etching promoted enamel
demineralization into deeper areas than was possible with
self-etching adhesives, thus increasing the micromechani-
cal interlocking of adhesives into the etched surface
through selective demineralization of the enamel
prisms.2,20,26 In this study, the failure mode of UBQ-PA
showed a high tendency to cause enamel fracture (E).
These features may explain why UBQ-PA showed signifi-
cantly higher μSBS than the other groups (p < 0.001). On
the other hand, there were no significant differences in
μSBS between GPB-0, GPB-10, and GPB-PA. Cohesive fail-
ure in the bond (B) occurred most frequently in GPB-PA
(27.3%), whereas the rate in UBQ-PA was 12.0%. Moreover,
the UTS test indicated that bond strength in GPB was sig-
nificantly lower than in UBQ. The UTS test method is consid-
ered a further valuable method for evaluating the mechani-
cal properties of bonding resins, resin composites, and
tooth structure.37 It was reported that the UTS of the bond-
ing resins was material- and storage-time dependent.31
From these results, low mechanical strength of GPB could
have affected the μSBS test in GPB-PA, resulting in cohe-
sive failure in the bonding resin.
Li et al15 found an ABRZ on enamel after application of
a 2-SEA that contained the functional monomer 10-MDP.
From their study, it was suggested that the formation of
the enamel ABRZ might be influenced by the presence of
the functional monomer. In self-etching adhesives, the
functional monomer behaves as an etchant, allowing the
resin monomer to penetrate into the demineralized
enamel and chemically interact with HAp crystals. This pro-
tects the crystals from acid attack and forms an ABRZ.15
We reported that funnel-shaped erosions were observed in
the adhesive-enamel interface of 1-SEA, despite the fact
that it contained 10-MDP. However, with selective phos-
phoric acid etching, the funnel-shaped erosion was not
observed.28 The ABRZ is thought to play an important role
in the prevention of recurrent caries, sealing of restoration
margins, and improving restoration durability.22 However,
funnel-shaped erosion was also detected in 1-SEAs, mean-
ing that there was an area susceptible to the acid-base
attack beneath the ABRZ. However, little has been re-
ported about the enamel ABRZ with the use of no-waiting
self-etching concept adhesives. In this study, the forma-
tion of the enamel ABRZ was confirmed in all adhesive
groups with or without selective phosphoric acid etching
(Fig 3). UBQ and GPB include 10-MDP, and this may con-
tribute to formation of the ABRZ. In UBQ-0, UBQ-10, GPB-
0, and GPB-10, the widths of the ABRZ were approximately
0.2 μm, and the funnel-shaped erosion or defect along the
interface was detected. There were some morphological
differences between UBQ and GPB. In contrast, in UBQ-0
and UBQ-10 (Figs 3a and 3b), the top area of the erosion
413
Sato et al
was wider than bottom observed in GPB-0 and GPB-10
(Figs 3d and 3e). The difference of composition or pH of
the adhesives might be the reason for these morphologi-
cal differences. Furthermore, the morphological features
were also influenced by the method of enamel etching.
Phosphoric acid etching prior to UBQ and GPB application
showed thicker ABRZ (approximately 3 μm) and no funnel-
shaped erosion was created at the adhesive-enamel inter-
face (Figs 3c and 3f), consistent with the results of other
studies.14,28 Since phosphoric acid etching is more ag-
gressive than self-etching, resin monomers penetrate
deeper into the etched layer.
UBQ and GPB contain 10-MDP as a functional monomer.
The ability of 10-MDP to readily establish an ionic bond with
HAp has been demonstrated.38 Previous studies have re-
ported that self-etching adhesives demineralize enamel
mildly, resulting in shallower intercrystallite resin infiltration
and less interprismatic resin tag formation, compared with
phosphoric acid-etching adhesives.6,9 From these reports,
GPB might demineralize enamel more deeply than UBQ. The
manufacturer indicates that UBQ includes a new hydrophilic
amide monomer which is more hydrophilic than HEMA. Con-
trary to this, GPB does not contain HEMA. It is likely that
the presence of hydrophilic monomers favors monomer pen-
etration into moist tooth substrates. This amide monomer
is a polyfunctional monomer in contrast to the monofunc-
tional monomer HEMA. The polyfunctional monomer might
provide a stronger polymer network (crosslinks), resulting in
higher UTS. UBQ includes sodium fluoride at reportedly the
same concentration as in 2-SEA, which was shown to influ-
ence the amount of dentin structure remaining after acid
challenge.13 Therefore, the adhesive-enamel interface might
be enhanced by the inclusion of sodium fluoride, which pos-
sibly accounts for the different widths of the funnel-shaped
erosion between UBQ and GPB. Further research is needed
to assess the influence of the sodium fluoride concentra-
tion at the adhesive-enamel interface.
A previous study demonstrated that the clinical perfor-
mance of 1-SEAs was clinically acceptable.36 However, the
interfacial morphologies observed in the current study re-
vealed that the funnel-shaped erosion was present in both
UBQ and GPB. It has raised doubts about the acid resis-
tance ability of these 1-SEAs on enamel. In addition, we
reported that 2-SEA without phosphoric acid etching formed
stable adhesive-enamel interfaces.28 Peumans et al,25 in
their 13-year clinical evaluation of composite restorations in
noncarious cervical lesions using 2-SEAs, reported that the
clinical efficacy remained excellent, with selective acid-etch-
ing of the enamel only having a minor positive effect. Aki-
moto et al1 reported that composite restorations with
2-SEAs demonstrated excellent results, with only minimal
incidence of marginal discoloration of the composite restor-
ation, which could be recovered by refurbishment (polishing)
at the recall appointment. These studies concluded that
phosphoric acid etching of the enamel may not be essential
for the 2-SEAs.
In this study, selective phosphoric acid etching improved
the bond strength of UBQ. In contrast, there was no signifi-
414
cant difference between GPB-0, GPB-10, and GPB-PA. There-
fore, the null hypothesis that the extension of application
time improves the adhesive-enamel interface was partially
rejected. The other two null hypotheses (selective phos-
phoric acid etching has no effect on the adhesive-enamel
interface when the no-waiting self-etching concept adhesive
is used, and all no-waiting self-etching concept adhesives
show the same trend) were rejected.
CONCLUSION
The μSBS results suggest that phosphoric acid etching of
enamel is material dependent. The enamel ABRZ was con-
firmed in all groups. However, selective phosphoric acid
etching created a stable adhesive-enamel interface, while
the no-waiting self-etching concept adhesive created thinner
ABRZ and exhibited erosion.
ACKNOWLEDGMENTS
This work was supported by the Japan Society for the Promotion of
Science 17K17119 and 15K11105.
REFERENCES
1. Akimoto N, Takamizu M, Momoi Y. 10-year clinical evaluation of a self-
etching adhesive system. Oper Dent 2007;32:3–10.
2. Buonocore MG. A simple method of increasing the adhesion of acrylic fill-
ing materials to enamel surfaces. J Dent Res 1955;34:849–853.
3. de Goes MF, Shinohara MS, Freitas MS. Performance of a new one-step
multi-mode adhesive on etched vs non-etched enamel on bond strength
and interfacial morphology. J Adhes Dent 2014;16:243–250.
4. De Menezes Oliveira MA, Torres CP, Gomes-Silva JM, Chinelatti MA, De
Menezes FC, Palma-Dibb RG, Borsatto MC. Microstructure and mineral
composition of dental enamel of permanent and deciduous teeth. Mi-
crosc Res Tech 2010;73:572–577.
5. Devarasa GM, Subba Reddy VV, Chaitra NL, Swarna YM. Self-etching ad-
hesive on intact enamel, with and without pre-etching. Microsc Res Tech
2012;75:650–654.
6. Di Hipólito V, de Goes MF, Carrilho MR, Chan DC, Daronch M, Sinhoreti
MA. SEM evaluation of contemporary self-etching primers applied to
ground and unground enamel. J Adhes Dent 2005;7:203–211.
7. Fejerskov O, Josephsen K, Nyvad B. Surface ultrastructure of unerupted
mature human enamel. Caries Res 1984;18:302–314.
8. Frankenberger R, Taschner M, Garcia-Godoy F, Petschelt A, Krämer N.
Leucite-reinforced glass ceramic inlays and onlays after 12 years.
J Adhes Dent 2008;10:393–398.
9. Grégoire G, Ahmed Y. Evaluation of the enamel etching capacity of six
contemporary self-etching adhesives. J Dent 2007;35:388-397.
10. Gwinnett AJ. Structure and composition of enamel. Oper Dent 1992;5:10–17.
11. Hosaka K, Nakajima M, Takahashi M, Itoh S, Ikeda M, Tagami J, Pashley
DH. Relationship between mechanical properties of one-step self-etching
adhesives and water sorption. Dent Mater 2010;26:360–367.
12. Huang XQ, Pucci CR, Luo T, Breschi L, Pashley DH, Niu LN, Tay FR. No-
waiting dentine self-etching concept-Merit or hype. J Dent 2017;62:54–63.
13. Kirihara M, Inoue G, Nikaido T, Ikeda M, Sadr A, Tagami J. Effect of fluo-
ride concentration in adhesives on morphology of acid-base resistant
zones. Dent Mater J 2013;32:578–584.
14. Li N, Nikaido T, Alireza S, Takagaki T, Chen JH, Tagami J. Phosphoric
acid-etching promotes bond strength and formation of acid-base resistant
zone on enamel. Oper Dent 2013;38:82–90.
15. Li N, Nikaido T, Takagaki T, Sadr A, Makishi P, Chen J, Tagami J. The role
of functional monomers in bonding to enamel: acid-base resistant zone
and bonding performance. J Dent 2010;38:722–730.
The Journal of Adhesive Dentistry

16. Marchesi G, Frassetto A, Mazzoni A, Apolonio F, Diolosà M, Cadenaro M, Di
Lenarda R, Pashley DH, Tay F, Breschi L. Adhesive performance of a multi-
mode adhesive system: 1-year in vitro study. J Dent 2014;42:603–612.
17. Matsui N, Takagaki T, Sadr A, Ikeda M, Ichinose S, Nikaido T, Tagami J. The
role of MDP in a bonding resin of a two-step self-etching adhesive system.
Dent Mater J 2015;34:227–233.
18. Mena-Serrano A, Kose C, De Paula EA, Tay LY, Reis A, Loguercio AD, Per-
digão J. A new universal simplified adhesive: 6-month clinical evaluation.
J Esthet Restor Dent 2013;25:55–69.
19. Mine A, De Munck J, Vivan Cardoso M, Van Landuyt KL, Poitevin A, Kuboki
T, Yoshida Y, Suzuki K, Van Meerbeek B. Enamel-smear compromises
bonding by mild self-etching adhesives. J Dent Res 2010;89:1505–1509.
20. Muñoz MA, Luque I, Hass V, Reis A, Loguercio AD, Bombarda NH. Immedi-
ate bonding properties of universal adhesives to dentine. J Dent 2013;41:
404–411.
21. Nikaido T, Ichikawa C, Li N, Takagaki T, Sadr A, Yoshida Y, Suzuki K, Tagami
J. Effect of functional monomers in all-in-one adhesive systems on forma-
tion of enamel/dentin acid-base resistant zone. Dent Mater J 2011;30:
576–582.
22. Nikaido T, Nurrohman H, Takagaki T, Sadr A, Ichinose S, Tagami J. Nano-
leakage in hybrid layer and acid-base resistant zone at the adhesive/den-
tin interface. Microsc Microanal 2015;21:1271–1277.
23. Noda Y, Nakajima M, Takahashi M, Mamanee T, Hosaka K, Takagaki T,
Ikeda M, Foxton RM, Tagami J. The effect of five kinds of surface treat-
ment agents on the bond strength to various ceramics with thermocycle
aging. Dent Mater J 2017;36:755–761.
24. Perdigão J, Sezinando A, Monteiro PC. Laboratory bonding ability of a
multi-purpose dentin adhesive. Am J Dent 2012 Jun;25:153–158.
25. Peumans M, De Munck J, Van Landuyt K, Van Meerbeek B. Thirteen-year
randomized controlled clinical trial of a two-step self-etching adhesive in
non-carious cervical lesions. Dent Mater 2015;31:308–314.
26. Rosa WL, Piva E, Silva AF. Bond strength of universal adhesives: A sys-
tematic review and meta-analysis. J Dent 2015;43:765–776.
27. Saikaew P, Chowdhury AF, Fukuyama M, Kakuda S, Carvalho RM, Sano
H. The effect of dentine surface preparation and reduced application time
of adhesive on bonding strength. J Dent 2016;47:63–70.
28. Sato T, Takagaki T, Matsui N, Hamba H, Sadr A, Nikaido T, Tagami J.
Morphological evaluation of the adhesive/enamel interfaces of two-step
self-etching adhesives and multimode one-bottle self-etching adhesives.
J Adhes Dent 2016;18:223–229.
Vol 20, No 5, 2018
Sato et al
29. Shimada Y, Kikushima D, Tagami J. Micro-shear bond strength of resin-
bonding systems to cervical enamel. Am J Dent 2002;15:373–377.
30. Shimada Y, Tagami J. Effects of regional enamel and prism orientation
on resin bonding. Oper Dent 2003;28:20–27.
31. Takahashi M, Nakajima M, Hosaka K, Ikeda M, Foxton RM, Tagami J.
Long-term evaluation of water sorption and ultimate tensile strength of
HEMA-containing/-free one-step self-etching adhesives. J  Dent 2011;
39:506–512.
32. Torii Y, Itou K, Nishitani Y, Ishikawa K, Suzuki K. Effect of phosphoric
acid etching prior to self-etching primer application on adhesion of resin
composite to enamel and dentin. Am J Dent 2002;15:305–308.
33. Tsuchiya H, Tsubota K, Iwasa M, Ando S, Miyazaki M, Platt JA. Influence
of adhesive application time on enamel bond strength of single-step self-
etching adhesive systems. Oper Dent 2010;35:77–83.
34. Tsuchiya S, Nikaido T, Sonoda H, Foxton RM, Tagami J. Ultrastructure of
the dentin-adhesive interface after acid-base challenge. J  Adhes Dent
2004;6:183–190.
35. Tyas MJ, Burrow MF. Adhesive restorative materials: a review. Aust Dent
J 2004;49:112–121.
36. Van Landuyt KL, De Munck J, Ermis RB, Peumans M, Van Meerbeek B.
Five-year clinical performance of a HEMA-free one-step self-etching adhe-
sive in noncarious cervical lesions. Clin Oral Investig 2014;18:1045–1052.
37. Van Meerbeek B, Peumans M, Poitevin A, Mine A, Van Ende A, Neves A,
De Munck J. Relationship between bond-strength tests and clinical out-
comes. Dent Mater 2010;26:100–121.
38. Yoshida Y, Nagakane K, Fukuda R, Nakayama Y, Okazaki M, Shintani H.
Comparative study on adhesive performance of functional monomers.
J Dent Res 2004;83:454–458.
Clinical relevance: The results of this study indicated
that selective phosphoric-acid etching is recommended
to create stable adhesive-enamel interfaces when using
no-waiting self-etching adhesives to enamel.
415