The Saudi Journal for Dental Research (2017) 8, 510

King Saud University

The Saudi Journal for Dental Research

www.ksu.edu.sa
www.sciencedirect.com

ORIGINAL ARTICLE

Eect of six month storage on microtensile bond

strength of new elective etching adhesive system on
dentin in self-etching or etch-and-rinse approach
Leonardo Colombo Zeidan 1, Andre Figueiredo Reis 1, Alessandra Cassoni 1,
Jose Augusto Rodrigues *

Dental Research and Graduate Studies Division, Department of Restorative Dentistry, Guarulhos University, Guarulhos, SP, Brazil

Received 24 May 2016; revised 20 June 2016; accepted 22 June 2016

Available online 5 July 2016

KEYWORDS Abstract The aim of this study was to evaluate the microtensile bond strength (lTBS) to dentin of
Tensile strength; an elective etching adhesive system applied in etch-and-rinse (ER) or self-etching (SE) mode after
Dental bonding/methods; 6-months of storage in water. Thirty-six caries-free, human third molars were collected and stored
Dentin-bonding agents; in a 0.1% thymol solution. Dentin surfaces were exposed by 600-grit silicon carbide paper and teeth
Universal adhesive were divided into six groups (n = 6), according to the adhesive systems: a 2-step SE system, Clearfil
SE Bond (CSE); a 1-step SE adhesive Adper Prompt L-Pop (LPOP); Scotchbond Universal applied
as a 1-step SE adhesive (SBU-SE) and applied as a 2-step ER adhesive (SBU-ER); and two 2-step
ER adhesives: Adper Single Bond Plus (SBP) and Optibond Solo Plus (OSP). Composite build-ups
were constructed with TPH3 and cured in three increments of 2 mm each. Specimens were sectioned
with a slow-speed diamond saw under water in X and Y directions to obtain bonded beams that
were tested to failure in tension at a crosshead speed of 1.0 mm/min after one week or 6 months
of storage in water. Statistical analyses were computed using Repeated-Measures ANOVA and
Fishers LSD Tests (a = 0.05). There were no significant differences between 1-week and
6-months. SBU-ER and SBUSE showed the highest lTBS values and statistically differed from
LPOP (Fishers LSD). The SBP, OSP, and CSE groups showed intermediary lTBS and did not dif-
fer statistically from SBU-ER, SBUSE or LPOP, which presented the lowest lTBS values. The use

* Corresponding author at: CEPPE, Universidade Guarulhos UnG, Praca Tereza Cristina, 229, Guarulhos, SP CEP 07023-070, Brazil. Fax:
+55 11 24641700, +55 11 24641758.
E-mail addresses: lzeidan@prof.ung.br (L.C. Zeidan), reisandre@yahoo.com (A.F. Reis), acassoni@prof.ung.br (A. Cassoni), jrodrigues@prof.
ung.br, guto_jar@yahoo.com.br (J.A. Rodrigues).
1
Address: CEPPE, Universidade Guarulhos UnG, Praca Tereza Cristina, 229, Guarulhos, SP CEP 07023-070, Brazil. Fax: +55 11 24641700,
+55 11 24641758.
Peer review under responsibility of King Saud University.

Production and hosting by Elsevier

http://dx.doi.org/10.1016/j.sjdr.2016.06.006
2352-0035 2016 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
6 L.C. Zeidan et al.
of elective etching adhesive system in dentin with the etch-and-rinse or self-etching approach did not
compromise the bond strength and showed stable bonds after six months of storage in water.
2016 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University. This is
an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
1. Introduction
Bonding to dental enamel is considered effective and usually
presents high bond strength values.20 Resin-dentin bonds are
more difficult to achieve than resin-enamel bonds, because
dentin bonding relies on organic components12 However,
adhesive systems should present similar performance on
enamel and dentin to increase the clinical performance.
Resin-dentin bonds are normally created by infiltration of
hydrophilic resin monomers into a previously demineralized
dentin matrix.1,21,22 An essential condition for the formation
of the hybrid layer is the maintenance of the dentin organic
matrix moist after demineralization, which supports the expan-
sion of the collagen fibrils and preserves the integrity of the
interfibrillar spaces. This disposition allows an appropriate
infiltration of the resin monomers dissolved in non-aqueous
organic solvents or an aqueous solution of hydrophilic
primers.16
Alternatively, the self-etch bonding technique uses acidic
monomers that combine tooth surface etching and priming
in a single procedure, reducing the risk of technical deficiencies
from etch-and-rinse systems.25 The advantage of the self-
etching adhesives (SEA) technique is the simultaneous dem-
ineralization and resin infiltration, which should lead to a thin
and an optimally infiltrated hybrid layer.19,21,24 Also, SEAs are
less aggressive than phosphoric acid used in the etch-and-rinse
technique.21 However, bonding to enamel is not as effective as
it is in dentin, and it is generally not indicated for use on
enamel surfaces without a prior phosphoric acid etching step
for direct or indirect restorative procedures, especially in
unground enamel.8,10 The selective enamel etching combined
with a mild pH SEA can therefore today be recommended
to achieve effective and durable bonding to enamel and
dentin.24,27
However, clinicians should not inadvertently etch dentin
walls in the selective etching process. If it occurs, the adhesive
may not be able to properly infiltrate etched dentin to produce
a good-quality hybrid layer. SEAs may have reduced perfor-
mance when bonding to etched dentin surfaces.11,19,23
In an attempt to overcome this problem, manufacturers
have developed the so-called Universal adhesives, that can
be applied in both self-etching or etch-and-rinse modes. How-
ever, few studies describe the use and the performance of such
adhesive systems.
The aim of this study was to evaluate the microtensile bond
strength (lTBS) of an elective etching adhesive system applied
in the etch-and-rinse and self-etching modes and the stability
after six months of water-storage. The null hypothesis of this
study is that the bond strength values produced by the elective
etching adhesive system do not differ from strictly etch-and-
rinse or the self-etching adhesives after seven days or six
months of storage in water.
2. Methods and materials
2.1. Experimental design
The factor under study was adhesive system in six levels
(n = 6), analyzed by repeated measurements at seven days
and six months in water storage. The experimental adhesive
groups were: SBU-SE- Scotchbond Universal (3M ESPE, St.
Paul, MN, USA) applied as a 1-step self-etch adhesive; SBU-
ER- Scotchbond Universal (3M ESPE, St. Paul, MN, USA)
applied as a 2-step etch-and-rinse adhesive, the etch-and-
rinse control groups were SBP- Adper Single Bond Plus (3M
ESPE, St. Paul, MN, USA), OSP- Optibond Solo Plus (Kerr,
Orange, CA, USA), and the control self-etching groups were
CSE- Clearfil SE Bond (Kuraray, Kurashiki, Japan), and
LPOP- Adper Prompt L-Pop (3M ESPE, St. Paul, MN,
USA) (Table 1). The dependent variable was microtensile bond
strength (lTBS) in MPa.
2.2. Specimens preparation
Thirty-six caries-free, human third molars were collected
according to the local Institutional Review Board (#
19/2009), with the informed consent of the donors. They were
stored in a 0.1% thymol solution at 4 C and used within one
month following extraction.
Prior to bonding procedures, the roots and occlusal enamel
were removed and the exposed middle dentin surfaces were
wet-polished with 600-grit silicon carbide paper under running
water to create a standard smear layer.5
2.3. Bonding procedures
For groups SBU-ER, SBP and OSP the 35% phosphoric acid
gel (ScotchBondTM Etchant, 3M ESPE, St. Paul, MN, USA)
was used to etch dentin surfaces for 15 s. Dentin was thor-
oughly rinsed with water and excess water was removed prior
to adhesive application. Adhesive was applied following man-
ufacturer directions (Table 1) and light-cured using an LED
(Radii Plus SDI, Victoria, Australia) with a power output
of 2000 mW/cm2. Subsequently, composite build-ups were
made incrementally (TPH3 Dentsply Caulk, Milford, DE,
USA) on each bonded surface. Three 2 mm increments were
individually placed and light-cured for 20 s.
2.4. Microtensile bond strength test
Restored teeth were stored in distilled water at 37 C for 7
days. Afterward, they were serially sectioned in both x
and y directions using a diamond saw (IsoMet 1000; Buehler
Ltd, Lake Bluff, IL, USA) under water cooling, resulting in
Microtensile bond strength of adhesive system on dentin 7

Table 1 Materials used in this study.

Material Components* Batch Manufacturers
Number recommended
protocols**
CSE- Clearfil SE Bond Self-etching primer: MDP, HEMA, DMA, catalyst, water Bond: e; f; g (10s)
(Kuraray, Kurashiki, Japan) Bonding: MDP, HEMA, DMA, Bis-GMA, 01657A
Filler, catalyst Primer:
01107A
SBU- Scotchbond Universal MDP Phosphate Monomer, Dimethacrylate resins, HEMA, 457855 Self-etch: e; f; g (10s)
(3M ESPE, St. Paul, MN, VitrebondTM Copolymer, Filler, Ethanol, Water, Initiators, Silane Etch-and-rinse: a
USA) (15s); b (15s); c; e; h; i
(10s)
SBP- Adper Single Bond Plus Bis-GMA, HEMA, dimethacrylates, methacrylated polyalkenoic acid, N318844 a (15s); b (10s); c; d; f;
(3M ESPE, St. Paul, MN, copolymer, initiators, water, ethanol and silane-treated silica g (10s)
USA) nanofillers
OSP - Optibond Solo Plus Bis-GMA, HEMA, GPDM, initiator, ethanol, fumed silica, barium 4312056 a (15s); b (10s); c; e
(Kerr, Orange, CA, USA) aluminoborosilicate and sodium hexafluorosilicate (15s); f; g (20s)
LPOP - Adper Prompt L-Pop Liquid 1 (red blister): Methacrylated phosphoric esters, Bis-GMA, 465223 d; f; g (10s)
(3M ESPE, St. Paul, MN, Initiators based on camphorquinone, Stabilizers
USA) Liquid 2 (yellow blister): Water, HEMA, Polyalkenoic acid, Stabilizers
TPH3 (Dentsply Caulk, Bis-GMA, Bis-EMA, TEG-DMA, TiO2, silica, Ba-B-F-Al-Si Glass L697363E g (20s)
Milford, DE, USA) Shade:
A3
ESPE Scotch Bond Etchant 35% phosphoric acid, silica 7523
(3M ESPE, St. Paul, MN,
USA)
*
Abbreviations: Bis-GMA: bisphenol-glycidyl methacrylate; HEMA: 2-hydroxyethyl methacrylate; DMA: dimethacrylate; GPDM: glycerol
phosphate dimethacrylate; UDMA: Urethane dimethacrylate; MDP: 10-methacryloyloxydecyl dihydrogen phosphate; PENTA: phosphonated
penta-acrylate ester; EDMAB: Ethyl 4-dimethyl amino benzoate.
**
Application techniques a: acid etching; b: rinsing; c: excess moisture removed from the preparation using a cotton pellet; d: application of
two consecutive coats of adhesive; e: application of one coat of the adhesive; f: gently air-drying; g: light-cure.

beams with a bonded cross-sectional area of approximately
1.0 mm2. Five beams for each tooth were tested at each evalu-
ation time. Beams were individually fixed to a custom-made
testing device with a cyanoacrylate adhesive (Loctite Super
Bond Gel; Henkel, Dusseldorf, Germany) and subjected to
tensile load at a crosshead speed of 1.0 mm/min until failure
(EZ Test, Shimadzu, Kyoto, Japan). Fractured specimens were
measured using a digital micrometer (Mitutoyo Co., Tokyo,
Japan) and lTBS values were expressed in MPa.
Failure mode was observed in a stereomicroscope at a 100
magnification (PanTec, Panambra Ind. e Tecnica SA, Sao
Paulo, Brazil) and scored according to one of three failure
modes: cohesive failure in dentin, adhesive failure between
dentin and composite and cohesive failure in composite resin.
Instead of classifying failures as mixed, the area percentage of
each type of failure in each fractured specimen was recorded.
2.5. SEM observation
Two beams of each group were sequentially polished, coated
with carbon (MED 010) and hybrid layer was observed under
a Scanning Electron Microscopy (LEO 440 VP). Representa-
tive areas of the interfaces were photographed (Figs. 27).
2.6. Statistical analysis
Five beams were tested for each tooth, and the mean bond
strength value was considered (n = 6). Bond strength values
were statistically analyzed by Repeated-Measures ANOVA,
followed by Fishers LSD Tests at the 95% confidence level
(IBM SPSS Statistics Version 20, IBM Company, Armonk,
NY, USA).
3. Results
No pre-test failures were recorded. The sample power observed
was 0.87 at one week and 0.86 at six months. There were no
significant differences between lTBS values observed at one
week and six months. Means lTBS and standard deviation
are shown in Table 2. Significant differences were observed
among tested adhesive groups (p < 0.05). The SBU-ER and
SBU-SE presented the highest lTBS means significantly higher
than LPOP according to Tukey test. The other groups pre-
sented intermediary lTBS values and did not differ from
SBU-ER, SBU-SE and from LPOP (Table 2).
The most frequent failures in all groups were cohesive in
resin, followed by adhesive failure. The groups SBP, CSE,
and LPOP showed no cohesive failure in dentin (Fig. 1).
4. Discussion
Adhesion of composite resins to dentin has been the main subject
of numerous studies. The wet bonding technique solves the
major problem of collagen collapse after acid-etching and results
in better resin penetration into acid etched dentin.11 However,
this technique has been reported to be complex and sensitive.7
Theoretically, SEAs are able to demineralize and infiltrate
dentin simultaneously.4 However, they are not as efficient in
enamel as in dentin and result in low bond strength values to
8 L.C. Zeidan et al.

Table 2 Means and standard deviations (sd) in MPa of

microtensile bond strengths, and Tukey test results for the
different adhesive systems.
Groups Mean at one Mean at six General Mean
week (sd) months (sd) (sd) Tukey HSD*
SBU-ER 68.3 (28.3) 63.9 (10.3) 66.1 (20.4) A
SBU-SE 63.2 (24.1) 62.1 (13.0) 62.6 (19.4) A
SBP 61.7 (16.0) 60.7 (15.7) 61.2 (15.4) AB
OSP 58.7 (10.9) 59.9 (9.7) 59.3 (9.5) AB
CSE 52.4 (22.1) 51.9 (13.5) 52.2 (12.3) AB
LPOP 36.5 (16.5) 38.4 (17.7) 37.4 (16.0) B
*
Different letters indicate statistically significant differences
(p < 0.05).

enamel. Consequently, the bond between enamel and resin

fails after some time of function in the mouth.9 Clinically,
the selective enamel etching technique may be reliable to create
microscopic roughness in enamel to provide the micromechan-
ical retention and avoid dentin over-etching. However, some
studies with SEAs have shown lower bond strengths when den-
tin is accidentally acid etched, because demineralized dentin is
not fully infiltrated by the SEAs.2,15 Since the balance between
dentin demineralization depth and extent of monomer penetra-
tion of the self-etching adhesives is the key to a good quality
interface between enamel and resin,18 elective etching or
universal adhesives have been developed.
As the elective etching adhesive system presented dentin
bond strength values similar to strictly etch-and-rinse and sim-
ilar or higher lTBS values than self-etching adhesives, the first
null hypothesis was partially accepted. In addition, as 6-
months storage in water did not produced any significant dif-
ference in bond strengths for all adhesives the second null
hypothesis was accepted.
Although, phosphoric acid can demineralize dentin more
deeply than a mild SEA, the tested elective etching adhesive
system (SBU) showed bond strength values compatible to
the etch-and-rinse adhesive systems (SBP and OSP) in both
modes of use, self-etching and etch-and-rinse, and are in agree-
ment with Perdigao et al., (2012)17 which found that SBU
groups ranked in the same statistical subset regardless of the
dentin treatment. Hanabusa et al. (2012)6 also found similar
results to another elective etching adhesive (G-Bond Plus) in
the immediate bond strength to dentin with self-etch or etch-
and-rinse approach. Also the experimental groups, SBU-ER
Figure 2 Representative SEM of a resin-dentin interface pro-
duced by Scotchbond Universal applied on etching and rinse
mode.
and SBU-SE, showed higher incidence of cohesive and adhe-
sive failures.
Mena-Serrano et al. (2013)13, found in a clinical trial of
Class V restorations that the behavior of the multimode adhe-
sive is not affected by the bonding strategy at 6 months. In the
present study, storage in water also did not affect bond
strengths, demonstrating stability of both self-etching (SE)
and etch-and-rinse (ER) approaches.
This elective etching adhesive system (SBU) presents a mild
pH (2.7), and a composition similar to other etch-and-rinse
systems available, which may explain the behavior similar to
the etch-and-rinse systems. However, it also presents the func-
tional monomer 10-MDP (10-methacryloyloxydecyl dihydro-
gen phosphate), which provides suitable self-etching
properties. The MDP monomer is also present in the CSE,
which showed bond strengths similar to SBU in both applica-
tion modes. MDP can chemically interact with hydroxyapatite
imparting better resistance toward degradation by prevention
of micro and nanoleakage.26
Recently, some researchers have described the presence of a
self-assembled nano-layering as a result from interaction of the

SBU-ER

SBU-SE

SBP Cohesive failure in resin

Adhesive failure
OSP
Cohesive failure in denn
CSE

LPOP

0% 20% 40% 60% 80% 100%

Figure 1 Percentage of failure modes in the experimental groups.

Microtensile bond strength of adhesive system on dentin
Figure 3 Representative SEM of a resin-dentin interface pro-
duced by Scotchbond Universal applied on self-etching mode.
Figure 4 Representative SEM of a resin-dentin interface pro-
duced by the etch and rinse adhesive Adper Single Bond Plus.
functional MDP, first with synthetic hydroxyapatite,28 fol-
lowed with enamel and dentin.5 This layer provides multi-
functional properties to the interface with, in particular, direct
benefit to bond durability.10 It seems that the strong
hydrophobic nature of the nano-layered structure may help
to protect the hybrid layer against biodegradation,1 improving
the protection of collagen against degradation, and making
residual hydroxyapatite more resistant to acidic dissolution.16
Both SBU and CSE adhesives may present such self-
assembled nano-layering of 10-MDP molecules, joined by
stable MDP-Ca salt formation, and development of a stronger
phase at the adhesive interface, which must also increase the
mechanical strength of the adhesive interface and may explain
the documented favorable bond strength of this adhesives and
clinical longevity of restorations.27
9
Figure 5 Representative SEM of a resin-dentin interface pro-
duced by the etch and rinse adhesive Optibond Solo Plus.
Figure 6 Representative SEM of a resin-dentin interface pro-
duced by the self-etching system Clearfil SE Bond.
However, CSE has demonstrated lower enamel bond
strength values than the conventional adhesive systems, both
when applied as self-etching or with previous acid-etching.3
Therefore, previous enamel etching did not improve enamel
bonding and also could not improve dentin bonding.3 Also,
the bond strength observed to SBU may be due to interaction
with other compounds as the VitrebondTM copolymer, which is
related to improve adhesion to moist and dry dentin and
results in some chemical bond to mineral phase of dentin
and enamel or due to the type and percentage of solvent which
could allow more penetration in etched dentin.14
SBU performed well in the self-etching (SBU-SE) and etch-
and-rinse (SBU-ER) modes and provides flexibility for the
clinician to choose one adhesive that presents dentin bond
10
Figure 7 Representative SEM of a resin-dentin interface pro-
duced by the self-etching adhesive Adper Prompt L-Pop.
strengths similar with conventional etch-and-rinse and self-
etching adhesive systems.
5. Conclusion
The in vitro use of an elective etching adhesive system in the
dentin with the etch-and-rinse or self-etching approaches did
not compromise bond strengths and presented stable results
after six months of storage in water.
Conict of interest
None declared.
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