d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600

available at www.sciencedirect.com

journal homepage: www.intl.elsevierhealth.com/journals/dema

Nanoleakage, ultramorphological characteristics, and

microtensile bond strengths of a new low-shrinkage
composite to dentin after artificial aging

Sillas Duarte Jr. ∗ , Jin-Ho Phark, Fabiana Mansur Varjão, Avishai Sadan
Department of Comprehensive Care, Case Western Reserve University, School of Dental Medicine, 10900 Euclid Avenue,
Cleveland, OH 44106-4905, USA

a r t i c l e i n f o a b s t r a c t

Article history: Objectives. To study the microtensile bond strengths and nanoleakage of low-shrinkage com-
Received 9 September 2008 posite to dentin. The null hypotheses tested were (1) aging does not affect the bonding
Accepted 31 October 2008 of low-shrinkage composite; (2) there is no difference in microtensile bond strengths and
nanoleakage using different bonding strategies.
Methods. 32 extracted molars were assigned to one of four groups: LS System Adhesive (LS, 3M
Keywords: ESPE); dentin etched for 15 s with phosphoric acid + LS System Adhesive (LSpa); Adper Single
Microtensile bond strengths Bond Plus (SB, 3M ESPE); SB + LS Bond (SBLS). Occlusal dentin was exposed and restored with
Nanoleakage Filtek LS (3M ESPE). The samples were tested after 24 h or after 20,000 thermocycles and 6
Dentin adhesion months of aging. Teeth were sectioned with a cross-section of 0.8 ± 0.2 mm2 and fractured at
Hybrid layer thickness a crosshead speed of 1 mm/min. The data were submitted to ANOVA/Duncan’s post hoc test,
Aging at p < 0.05. Five slabs from each group were selected and immersed in 50 wt% ammoniacal
Silorane silver nitrate. Then, specimens were processed for SEM, the silver penetration was measured
Scanning electron microscopy and data analyzed with Kruskal–Wallis at p < 0.05.
Results. No statistically significant difference was found among the experimental groups for
the factor dentin treatment (p = 0.165) and aging (p = 0.091). All experimental groups exhibit
some degree of nanoleakage. There was no adhesion of Filtek LS applied directly over dentin
surfaces treated with SB.
Significance. The new low-shrinkage resin composite showed compatibility only with its ded-
icated adhesive. Pre-etching did not improve the bond strengths to low-shrinkage resin
composite. Some degree of nanoleakage was evident in all groups.
© 2008 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

1. Introduction Recently, a new category of resin matrix for dental com-

Despite these improvements, methacrylate-based composites
are limited, however, especially with respect to polymeriza-
tion shrinkage and marginal integrity. Impairment of marginal
integrity can produce microleakage [1], post-operative sensi-
tivity [2], and debonding [3], all reducing the longevity of the
restoration.
posite was developed based on ring-opening monomers
[4]. This hydrophobic composite derives from the combina-
tion of siloxane and oxirane, thus the name silorane [5].
The major advantages of this innovative restorative mate-
rial are its reduced shrinkage and its mechanical properties
comparable to those of the methacrylate based composites
[5–7].

∗
Corresponding author. Tel.: +1 216 368 6737.
E-mail address: sillas.duarte@case.edu (S. Duarte Jr.).
0109-5641/$ – see front matter © 2008 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.dental.2008.10.012

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590 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600
A dedicated adhesive was produced by the manufac-
turer in order to obtain an appropriate bond of silorane
composite to dental hard tissues, especially because differ-
ences in silorane curing chemical mechanisms compared
to methacrylate composites yield superior adaptation. The
silorane adhesive is composed of a hydrophilic one-step
self-etch primer and a hydrophobic viscous bond coating
resin. The manufacturer claims that the one-step self-etch
choice is based on the increase popularity of this category
of adhesives. However, one-step self-etch adhesives have
limitations when bonding to dentin [8–10], and their long-
term bond still is undetermined [11–15]. Some questions
remain about the bonding abilities of this dedicated adhe-
sive. Although this new composite can form a strong bond
with identical material, its capacity to form bonds with dis-
similar materials is still open to question. If the silorane
composite resin can adhere to methacrylate-based adhesive,
it raises the subject as about how to improve or at least to
maintain acceptable bond strengths and levels of nanoleak-
age.
This project studied the microtensile bond strengths (␮TBS)
and nanoleakage in the dentin hybrid layer of low-shrinkage
composite, as a function of time and dentin treatment.
The null hypothesis is two-fold: (1) aging does not affect
either microtensile bond strengths or nanoleakage for low-
shrinkage composite; (2) there is no difference in bonding
of low-shrinkage composite using different bonding strate-
gies.
2. Material and methods
Thirty-two freshly extracted caries-free human third molars
were selected for the study and stored in 0.5% chloramine
solution (Fisher Scientific, Pittsburg, PA, USA) at 4 ◦ C for up to
one month after extractions. The teeth were scaled, cleaned,
and stored in distilled water for 24 h. The teeth were ran-
domly assigned to four experimental groups as shown in
Table 1.
Middle dentin was be exposed by sectioning the crowns
parallel to the occlusal surface with a precision low-speed
diamond saw (Isomet 1000, Buehler, Lake Buff, IL, USA),
under distilled water cooling. A dentin standard smear layer
was created by polishing the occlusal surface with 600-grit
SiC sandpaper for 60 s [16]. The bonded interface was pre-
pared according to the experimental groups (Table 1). After
adhesive application the clinical crowns were restored with
low-shrinkage composite Filtek LS (Lot: 7BB, 3M ESPE, St. Paul,
MN, USA) in 3 increments of 2.0 mm each. Each increment was
light-cured for 40 s (XL2500, 3M ESPE) at 0.5 mm curing dis-
tance and light intensity of 660 mW/cm2 constantly monitored
with a radiometer.
After bonding, each group was further divided into 2 sub-
groups (n = 4) to be tested for 24 h and after artificial aging. The
first half sub-group specimens were stored for 24 h and then
tested for microtensile bond strengths and nanoleakage. The
remaining specimens were tested after 6 months storage in
distilled water at 37 ◦ C and artificial aging by thermal cycling
with 20,000 cycles at 5 ◦ C and 60 ◦ C with 15 s dwell time and
7 s transfer time.
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2.1. Microtensile bond strengths (TBS)
After each aging period (24 h and 6 months + thermocycling),
specimens were retrieved from the storage medium for ␮TBS.
Specimens were sectioned parallel to the adhesive interface
to obtain 0.8 ± 0.1 mm thick slabs. Each slab was attached to
a phenolic ring and a second set of 0.8 ± 0.1 mm thick sec-
tions was obtained perpendicular to the adhesive interface.
Samples were measured with a caliper to assure proper fit
into the testing jig. The specimens were tested individually
by attaching them to a microtensile jig using cyanoacrylate
glue (ZapIt, DVA, Corona, CA, USA) [17]. The 0.8 mm × 0.8 mm
sticks were then submitted to a tensile load using a universal
testing machine (Model 1125, Instron Co., Canton, MA, USA)
at 1.0 mm min−1 cross-head speed. A digital caliper (Mitu-
toyo digital calipers; Mitutoyo Corp., Kanogawa, Japan) with
an accuracy of 0.001 mm was used to measure the sides of the
bonding interface and to calculate the bonding area in square
millimeters. The load in Newtons and the bonding surface
area of the specimen were registered using specialized soft-
ware (Test Works 4, MTS Nano Instruments, Oak Ridge, TN,
USA), and microtensile bond strengths calculated in MPa. Pre-
testing failures or spontaneous debonding were counted as
0 MPa.
Statistical analysis was performed with statistical software
(SPSS 15; SPSS Inc., Chicago, IL, USA). A 2-way analysis of vari-
ance (ANOVA) for dentin treatment and aging was computed,
followed by a Duncan’s post hoc test (p < 0.05).
2.2. Nanoleakage
Five resin-bonded dentin slabs from each experimental group
were used for nanoleakage evaluation. The specimens were
coated with two layers of nail polish, except for a 1.0 mm rim
around the restoration, to allow contact of the tracing agent
with the margins of the restoration. In order to avoid artifacts
due to desiccation, the specimens were rehydrated by immers-
ing in distilled water for 20 min before immersion in the tracer
solution [18]. Then the specimens were immersed in a aque-
ous solution of 50 wt% ammoniacal silver nitrate (pH 9.5) for
24 h, followed by 8 h in a photo-developing solution, in order to
permit the reduction of diammine silver ions to metallic silver
grains [19].
The specimens were retrieved from the photo-developing
solution and washed in running water for 1 min. The nail pol-
ish was removed and the specimens immediately immersed
in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer at pH
7.4 for 12 h at 4 ◦ C. After fixation, the specimens were rinsed
three times with 20 mL of 0.2 M sodium cacodylate buffer at
pH 7.4 for 1 h, followed by distilled water for 1 min. The speci-
mens were dehydrated in ascending concentrations of ethanol
as follows: 25% for 20 min, 50% for 20 min, 75% for 20 min, 95%
for 30 min, and 100% for 60 min.
After the final ethanol step, the specimens were met-
allurgically polished to high gloss with waterproof silicon
carbide papers of decreasing abrasiveness (600-, 800- and
1200-grit) followed by soft cloth with increasingly fine dia-
mond suspensions of 2 ␮m and 1 ␮m (Buehler Ltd.). To remove
any residual debris the specimens were sonicated in 100%
ethanol for 10 min, thoroughly dried, and demineralized in
 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600 591

Table 1 – Restorative techniques used.

Group Material Composition (Lot)
Self-etch primer: Phosphorylated
LS methacrylates, vitrebond
LS Adhesive copolymer, bisphenol A
diglycidylmethacrylate
(BisGMA),
2-hydroxyethylmethacrylate
(HEMA), water, ethanol,
silane-treated silica filler,
initiators, stabilizers (Lot: 7AD)
LS Bond: Hydrophobic
LSpa methacrylates, phosphorylated
methacrylates, triethylene
glycol dimethacrylates
(TEGMA), silane-treated silica
filler, initiators, stabilizers (Lot:
7AC)
Adper Single Bond Plus: BisGMA,
Adper Single Bond HEMA, dimethacrylates,
SBLS
Plus + LS Bond ethanol, water, photoinitiator
system, methacrylate
functional copolymer of
polyacrylic and polyitaconic
acids, 5 nm silica particles (Lot:
7KL)
LS Bond: Hydrophobic
methacrylates, phosphorylated
methacrylates, triethylene
glycol dimethacrylates
(TEGMA), silane-treated silica
filler, initiators, stabilizers (Lot:
7AC)
Manufacturer
3M ESPE, St. Paul, MN, EUA
3M ESPE, St. Paul, MN, EUA
Restorative technique
1. Apply 1 coat of the self-etch
primer for 15 s with gentle
agitation using fully saturated
applicator. Gently air thin to
evaporate solvent and obtain
an even film. Light-polymerize
for 10 s
2. Apply the Bond to the entire
preparation using fully
saturated applicator. Gently air
thin until the Bond is spread to
an even film and does not
move any longer.
Light-polymerized for 10 s
1. Apply Scotchbond Etchant
(35%) to dentin. Wait 15 s and
rinse for 10 s. Blot excess water
using cotton pellet
2. Apply 1 coat of the self-etch
primer for 15 s with gentle
agitation using fully saturated
applicator. Gently air thin to
evaporate solvent and obtain
an even film. Light-polymerize
for 10 s.
3. Apply the Bond to the entire
preparation using fully
saturated applicator. Gently air
thin until the Bond is spread to
an even film and does not
move any longer.
Light-polymerized for 10 s
1. Apply Scotchbond Etchant
(35%) to dentin. Wait 15 s and
rinse for 10 s. Blot excess water
using cotton pellet.
2. Immediately after blotting,
apply 2–3 consecutive coats of
adhesive for 15 s with gentle
agitation using fully saturated
applicator. Gently air thin for
5 s to evaporate solvent.
Light-polymerize for 10 s
3. Apply the LS Bond to the
entire preparation using fully
saturated applicator. Gently air
thin until the Bond is spread to
an even film and does not
move any longer.
Light-polymerized for 10 s

Adper Single Bond BisGMA, HEMA, 1. Apply Scotchbond Etchant

SB dimethacrylates, ethanol, 3M ESPE, St. Paul, MN, EUA
Plus (35%) to dentin. Wait 15 s and
water, photoinitiator system, rinse for 10 s. Blot excess water
methacrylate functional using cotton pellet
copolymer of polyacrylic and 2. Immediately after blotting,
polyitaconic acids, 5 nm silica apply 2–3 consecutive coats of
particles (Lot: 7KL) adhesive for 15 s with gentle
agitation using fully saturated
applicator. Gently air thin for
5 s to evaporate solvent.
Light-polymerize for 10 s.

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592 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600

0.5% silica-free phosphoric acid for 1 min. Specimens were

then dried by immersion in hexamethyldisilazane (Electron
Microscope Sciences, Fort Washington, PN, USA) for 10 min,
placed on filter paper inside a covered glass vial, and air dried
at room temperature [20]. The specimens mounted in alu-
minum stubs, sputter-coated with carbon and observed under
a field emission scanning electron microscope (Fe-SEM) with
a backscattered detector (S-4500, Hitachi High Technologies
America, Inc., Pleasanton, CA, USA) at an accelerating voltage
of 8.0 kV. The silver penetration along the preparation wall was
analyzed with the Quartz PCI 4.00 software (Quartz Imaging
Corporation, Vancouver, BC, Canada).
The silver penetration was analyzed according to differ-
ent areas of the bonded surface in: (1) bottom of hybrid layer,
(2) within the hybrid layer and (3) water-trees. The bottom
of hybrid layer was scored as: score 0: no leakage, score 1: Fig. 1 – Silver penetration for different experimental groups
intermittent leakage, and score 2: continuous leakage. Silver restored with silorane composite before and after aging.
penetration within the hybrid layer was graded as: score 0,
no silver deposit; score 1, slight silver deposits covering less
than 1/3 of HL thickness and score 2, heavy silver deposits
covering more than 1/3 of HL thickness. Water-trees at hybrid
paper inside a covered glass vial, and air-dried at room tem-
layer/adhesive layer interface were graded as: 0, absent and
perature [20].
1, present. The data were submitted to a Kruskal–Wallis non-
The restored specimens were embedded in self-curing
parametric statistical analysis at p < 0.05 (SPSS 15.0, SPSS Inc.,
epoxy resin (Buehler Ltd.) and stored at room temperature
Chicago, IL, USA).
under a hood for 18 h. After curing, the surface of each epoxy
cast was metallurgic polished with wet silicon carbide papers
2.3. Hybrid layer thickness of decreasing abrasiveness (up to 1200-grit) and then with an
increasingly fine diamond suspension to a particle size of 1 ␮m
Thirty extracted human third molars, refrigerated in a solu- (Buehler Ltd.). The top of the epoxy cast was protected with
tion of 0.5% chloramine for up to one month after extraction, non-adhesive tape and sectioned with a diamond saw to sep-
were used. The occlusal enamel was removed and thirty arate the specimen from the epoxy block. The specimens were
dentin disks parallel to the occlusal surface with a thickness sonicated in 100% ethanol for 5 min, thoroughly dried, dem-
of 800 ± 200 ␮m were obtained from middle dentin by slow- ineralized in 6N HCl for 30 s, and deproteinized in 2% NaOCl
speed sectioning with a precision diamond saw (Isomet 1000, for 10 min.
Buelher). A standard smear layer was created on the occlusal All specimens were mounted on aluminum stubs and
surface by wet sanding with 600-grit SiC sandpaper for 60 s sputter-coated with carbon. The bonded interface was
[16]. The dentin discs were restored according to the exper- observed under high resolution dual beam FESEM/focused
imental groups (Table 1), except for SB groups because of ion beam microscope at an accelerating voltage of 4.0–5.0 kV
complete debonding of the restoration. The specimens were (xT Nova Nanolab 200, FEI Company, Hillsboro, OR, USA).
fixed with glutaraldehyde and then dehydrated in ascending The hybrid layer thickness was analyzed with the Fei-
concentrations of ethanol. After the final ethanol step, the Navigator software (FEI Company). A 1-way analysis of
specimens were dried by immersion in hexamethyldisilazane variance (ANOVA) was computed followed by a Duncan post
(Electron Microscope Sciences) for 10 min, placed on a filter hoc test (p < 0.05).

Table 2 – Mean microtensile bond strengths (MPa), standard deviation, pre-testing failures/spontaneous debonding (PTF),
and number of specimens (N).
Time Dentin treatment Mean (SD) PTF N

LS Adhesive 61.8 (38.7)A* 9 52

Etch + LS Adhesive 73.7 (22.8)A 0 50
24 h
Single Bond + LS Bond 77.1 (24.3)A 0 62
Single Bond 4.4 (5.1)B 22 33

LS Adhesive 54.8 (38.4)A 11 45

Etch + LS Adhesive 71.6 (9.5)A 0 40
Aged
Single Bond + LS Bond 69.9 (19.2)A 0 38
Single Bond 0 (0)B 40 40

*
Means with same superscript uppercase letters are not significantly different (p < .05). Duncan’s test uses harmonic mean of group sizes because
group sizes are unequal.

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 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600 593

Fig. 2 – (A) Backscattered FESEM of silver deposits (white arrows) within the hybrid layer for LS adhesive after 24 h.
Magnification = ×2500. (B) Higher magnification of (A) showing nanoleakage within (white circle) and at the bottom of HL.
Note silver penetration within the resin tags (white arrow). Magnification = ×5000. (C) Backscattered FESEM of silver deposits
(white arrows) within the hybrid layer for LS adhesive after aging. Observe the presence of water-trees at the adhesive layer
(white circle) and silver deposits at the intervening zone (white arrow). Magnification = ×2500. (D) Higher magnification of
(C) showing water-trees (white circle), nanoleakage within the hybrid layer (white arrow), and silver deposits at the
intervening zone. Magnification = ×5000. C, Composite; A, adhesive layer; B, bond layer; HL, hybrid layer; RT, resin tags.

2.4. Ultramorphological study of dentin treated with
silorane self-etch primer
Six extracted human third molars, refrigerated in a solution
of 0.5% chloramine for up to one month after extraction, were
used. The occlusal enamel was removed with an Isomet 1000
diamond saw (Buehler Ltd.), and six dentin disks with a thick-
ness of 800 ± 200 ␮m were obtained from middle dentin by
slow-speed sectioning. A transverse groove was made in one
side of the dentin disks to facilitate splitting.
A standard smear layer was created on the occlusal surface
by wet sanding with 600-grit SiC sandpaper for 60 s. The self-
etching primer was applied according to the manufacturer’s
instructions and after acid etching dentin for 15 s. The dentin
discs were immersed in 100% acetone for 5 min and imme-
diately fixed, dehydrated, and dried for SEM evaluation, as
described above. After drying, the disks were fractured by care-
fully applying bending force on the groove of the non-etched
surface. Half of each disk was mounted to observe dentinal
tubules in cross-section and the other half to observe dentin
tubules longitudinally. All specimens were mounted on alu-
minum stubs and sputter-coated with carbon. The effect of
dentin etching was observed under high resolution dual beam
FESEM/focused ion beam microscope at an accelerating volt-
age of 4.0–5.0 kV and from magnification of 1000× to 40,000×.
3. Results
3.1. Microtensile bond strengths
The total-etch methacrylate-based dentin adhesive (SB) was
unable to produce sufficient bond strengths to hold the
silorane composite resin in place. Most of the specimens spon-
taneously debonded after 24 h and all restorations debonded
after water storage or during thermocycling. Thus, these
groups were removed from the statistical analysis since there
was no adhesion of Filtek LS applied directly over dentin sur-
faces treated with Single Bond Plus.
Two-way ANOVA (Table 2) performed for LS, LSpa, and
SBLS showed no statistical difference among the means for
“dentin treatment” (p = 0.165) and “aging” (p = 0.091). The high-
est mean bonds strengths were obtained when dentin was
pre-treated with phosphoric acid. No pre-testing failures were

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594 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600

Fig. 3 – (A) Backscattered FESEM of silver deposits within the hybrid layer for LSpa adhesive after 24 h.
Magnification = ×2500. (B) Higher magnification of (A) showing nanoleakage within and at the bottom of HL (white circle).
Note the presence of water-trees at the adhesive layer (white arrow). Magnification = ×5000. (C) Backscattered FESEM of
silver deposits (white arrows) within the hybrid layer for LSpa after aging. Observe the presence of water-trees at the
adhesive layer, silver deposits at the intervening zone and within the HL. Magnification = ×2500. (D) Higher magnification of
(C). Magnification = ×5000. A, Adhesive layer; B, bond layer; HL, hybrid layer; RT, resin tags.

observed for LSpa and SBLS at both 24 h and after aging. The
effects of dentin treatment versus aging were not statistically
significant (p = 0.485). Aging did not affect bond strengths of
different dentin treatments for silorane composite. The Dun-
can’s test showed no differences among LS, LSpa, and SBLS
groups (p = 0.410). The slightly decrease of bond strengths for
LS, LSpa, and SBLS after aging was not significantly different
from non-aged specimens.
3.2. Nanoleakage
It was not possible to observe the nanoleakage for SB group,
before and after aging, because of complete debonding of the
restorations. Silver penetration length for each experimental
group restored with silorane composite with shown in Fig. 1.
No statistical difference was found for nanoleakage at the
base of hybrid layer for LS, LSpa, and SBLS before and after
aging (p = 0.764). However, enough evidence was found to con-
clude that there were significant statistical differences among
the tested groups based on the silver penetration within the
hybrid layer (p = 0.003). Thus multiple comparisons were per-
formed using the Mann–Whitney U test. LS adhesive produced
less mean nanoleakage within the hybrid layer than occurred
in other experimental groups (p = 0.008). However, no statis-
tically differences were found in LS adhesive nanoleakage
before and after aging (p = 0.31). LSpa and SBLS presented sim-
ilar nanoleakage before and after aging.
All tested groups exhibited water-trees protruding from the
adhesive interface toward the adhesive layer (Figs. 2–4). No
statistical difference was found among the groups (p = .066).
Incomplete resin infiltration within the exposed collagen
fibrils was found for all specimens. Areas of small and dis-
continuous silver clusters were found within the hybrid layer
for all tested groups. Reticular nanoleakage was found for all
experimental groups. Reticular pattern occurred where water
was not totally removed in the adhesive layer or within the
collagen fibrils. Aging of the specimens showed an increase of
reticular pattern of nanoleakage, most probably where areas
of hydrolytic degradation occurred. Unprotected demineral-
ized dentin could absorb water leading to more silver uptake.
Unpolymerized resin present within the interfibrillar spaces
could generate a spotted pattern of nanolekage. Spotted pat-
terns were mostly observed for the specimens where the
dentin was acid-etched with phosphoric acid (Figs. 3 and 4).
In particular, SBLS after aging revealed an extensive spotted
silver deposit throughout the hybrid layer (Fig. 4C and D).

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 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600 595

Fig. 4 – (A) Backscattered FESEM of silver deposits within the hybrid layer for SBLS adhesive after 24 h.
Magnification = ×2500. (B) Higher magnification SBLS showing nanoleakage within and at the bottom of HL (white arrow).
Magnification = ×5000. (C) Backscattered FESEM of silver deposits within the hybrid layer, resin tags, and at the adhesive
layer for SBLS after aging. Observe the intense silver uptake. Magnification = ×2500. (D) Higher magnification of (C) revealing
the water-trees (white arrow) and punctuate nanoleakage within the resin globules and blister both at the adhesive layer
(white circle). Magnification = ×5000. C, Composite; A, adhesive layer; B, bond layer; HL, hybrid layer; RT, resin tags.

Reticular silver deposits were reduced in frequency and subse- third subset. LS Adhesive showed few resin tags and hybrid
quently were replaced by the spotted pattern of nanoleakage layer thickness raging from 1.7 to 2.0 ␮m (Fig. 5A and B). Acid-
throughout the hybrid layer. etching dentin before the application of LS Adhesive increased
Aging also affected the adhesive layer of most specimens the thickness of the hybrid layer (4.1–5.1 ␮m) and also the
(Figs. 2C and D, 3C and D, 4C and D). An increase in the number amount of resin tags (Fig. 5C and D). In addition, some filler
of water-trees was observed. However, it was not statistically from the low-shrinkage composite penetrated the resin tags.
significant. Microphase separation of copolymers also was SBLS showed a classic hybrid layer for total-etch adhesive
identified for the group SBLS after aging resulting in blister (Fig. 5E and F). However, a separation line between the Sin-
or resin globules with spotted nanoleakage pattern within the
adhesive layer (Fig. 4D). Aging also showed an increase of sil-
ver deposits at the tubuli walls for all tested groups, probably
because of hydrolytic degradation during thermo cycling and Table 3 – Mean thickness of the hybrid layer (␮m) and
storage. standard deviation.
Time Dentin treatment Mean (SD)
3.3. Hybrid layer thickness LS Adhesive 1.7 (0.23)C*
24 h Etch + LS Adhesive 5.1 (0.2)A
One-way ANOVA (Table 3) for LS, LSpa, SBLS showed statis- Single Bond + LS Bond 5.4 (0.08)A
tical differences for thickness of the hybrid layer (p < 0.001). LS Adhesive 2.0 (0.18)C
Different dentin treatments resulted in differences in the Aged Etch + LS Adhesive 4.1 (0.82)B
hybrid layer thickness. The Duncan’s test grouped the dentin Single Bond + LS Bond 4.1 (1.15)B
treatment and aging into three subsets. LSpa and SBLS were
*
Means with same superscript uppercase letters are not signifi-
grouped in the highest subset, followed by LSpa Aged and
cantly different (p < .05).
SBLS aged in the second subset, and LS and LS aged in the

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596 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600

Fig. 5 – (A) Secondary electron FESEM of specimen restored with LS. Note the few resin tags and the clear distinction
between the adhesive layer (A) and bond layer (B). Magnification = ×10,000. (B) The distinction between the LS hybrid layer
(HL) and adhesive layer (A) can be identified only in higher magnification. Magnification = ×40,000. (C) Secondary electron
FESEM of specimen restored with LSpa. Note the increase on resin tags compared to (A) and also the anastomosis among
the resin tubules. Magnification = ×3500. (D) Higher magnification of (C) revealing the hybrid layer created at the dentin
tubules also tubules but at and the penetration of the adhesive forming the resin tags. Magnification = ×10,000. (E)
Secondary electron FESEM of specimen restored with SBLS. Note the the clear distinction between the adhesive layer (A) and
bond layer (B). Magnification = ×5000. (F) Higher magnification of a specimen restored with SBLS revealing the hybrid layer
and resin tag. Magnification = ×25,000. R, Resin composite; A, adhesive layer; B, bond layer; HL, hybrid layer; RT, resin tags.

gle Bond Plus adhesive layer and the LS Bond was clearly 3.4. Ultramorphological evaluation
visible (Fig. 5E). Aging reduced the hybrid layer thickness for
all groups, however only LSpa was statistically significant The application of the self-etch primer (LS Self-etch primer,
(p < 0.001). 3M ESPE) on dentin produced an intense intertubular decalci-

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 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600 597

Fig. 6 – (A) Secondary electron FESEM of dentin treated with LS self-etch primer. Note the intense intertubular
decalcification diplaying the collagen fibers, however the tubules still are blocked with smear layer/smear plugs.
Magnification = ×8000. (B) Cross-sectional view of dentin treated with LS self-etch primer revealed a decalcification depth
from 1 to 1.7 ␮m. Note the presence of the smear plug. Magnification = ×25,000. (C) Secondary electron FESEM of dentin
etched with 35% phosphoric acid for 15 s followed by the application of the LS self-etch primer. Note the intense collagen
network exposure. Magnification = ×8000. (D) Cross-sectional view of dentin treated with LSpa revealed a decalcification
depth from 1.8 to 5.2 ␮m. Note the undetermined collagen at the basal portion of decalcified dentin (white arrow).
Magnification = ×25,000. ID, Intertubular dentin; PD, peritubular dentin; T, tubules; SP, smear plug.

fication, resulting in an exposed collagen network (Fig. 6A).

A decalcification depth of 1–1.7 ␮m was observed for inter-
4. Discussion
tubular dentin associated with a small and incomplete
This study showed that silorane dedicated adhesive can gen-
opening of the dentin tubules (Fig. 6B). The self-etch primer
erate acceptable microtensile bond strengths to dentin, but
“mild” pH was unable to completely remove the smear
that nanoleakage still is a problem. Acid etching dentin
layer, thus the tubules were mostly blocked by smear
improved the bond strengths for LS adhesive. The appli-
plugs.
cation of a total-etch methacrylate-based dentin adhesive
Acid-etching dentin associated with the application of the
followed by silorane hydrophobic bond resin (LS Bond, 3M
self-etch primer resulted in smear layer removal. Very intense
ESPE) produced sufficient microtensile bond strengths, but
intertubular microporosity with exposure of the outer per-
with significant nanoleakage, especially after aging. Silorane
itubular collagen was observed (Fig. 6C). The longitudinal view
composite applied directly over the dentin bonded with SB
of the etched dentin revealed a funnel shape etching pat-
resulted in complete debonding of the restorations. Earlier
tern with depth of demineralization raging from 1.8 to 5.2 ␮m
tests using a different total-etch acetone- and methacrylate-
(Fig. 6D). Undermined collagen was observed at the basal part
based dentin adhesive showed similar behavior when bonded
of the decalcified dentin among tubules with collagen fibrils
to silorane [21]. No correlation was found between microten-
protruding toward the tubule walls (Fig. 6D). The deepest areas
sile bond strengths and nanoleakage. In addition no interface
of etching were found around the tubules, whereas the shal-
gaps were found for all tested groups. Therefore the first null
lowest ones were found at the intertubular dentin among the
hypothesis was accepted: aging does not affect bond strengths
tubules.

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598 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600
or nanoleakage. Because the bond strategy plays an important
role in the adhesion of silorane-based composite to dentin, the
second hypothesis was rejected.
LS adhesive can be considered a “mild” self-etch adhesive
with a pH of 2.7, which produces intense intertubular micro-
porosity with a residual smear layer on the dentin surface
and preservation of the smear plugs. SEM evaluation showed
a discernible segregation among the hybrid layer, adhesive
layer, and bond layer. Our ultrastructural evaluation contra-
dicts other SEM study, but agrees with a recent micro-Raman
evaluation [12]. Few resin tags were observed for LS and the
presence of the smear plug within the dentin tubules prevents
proper resin infiltration. However, resin tags do not influence
the bond strengths of one-step self-etch adhesive [22]. Thus,
resin monomers must micromechanically interlock with the
exposed collagen network of dental structures in order to
promote adequate bond strengths. LS adhesive was able to
maintain satisfactory microtensile bond strengths before and
after aging. Short-term observations indicate that the bond
strengths are not correlated with the thickness of the hybrid
layer [23]. Conversely, silver uptake was found throughout
the hybrid layer for LS adhesive, indicating porosity within
this layer. The water present in the LS self-etch primer reacts
with the phosphorylated methacrylate, generating hydrogen
ions necessary to demineralize the dentin. However, areas
of incomplete polymerization or hydrogels can be formed if
water remains within the collagen network or at the adhe-
sive layer [24]. Uncured resin monomers from the adhesive can
trigger an osmotic pressure gradient from the dentin tubules,
degrading the bond after aging [25]. Water-trees were found
extending orthogonally from the hybrid layer into the adhesive
layer, especially after aging. Our findings contradict a recent
study in which water-trees were not observed within the silo-
rane adhesive layer [13]. Therefore, because one-step self-etch
adhesives can act as permeable membranes [25,26], the appli-
cation of a hydrophobic coating layer is essential to improve
the durability of the bonding [27]. However, some water-trees
also were found at the adhesive layer/bond layer interface.
An interesting study using micro-Raman identified spectra
of an intervening oxygen inhibition zone of 1 ␮m between
the cured primer surface and the bond layer [12]. Hypotheti-
cally, no oxygen inhibition layer should exist on the surface of
the composite because of the ring-opening polymerization of
the silorane molecule [28]. Futhermore, an oxygen inhibition
layer has only a moderate effect on bond efficacy of self-etch
adhesives [29]. The bond between successive layers of silo-
rane depends upon the chemical reactivity of the material,
and chemical reactivity lessens over time [28]. In our study,
the intervening zone also was recognized, particularly after
aging. The silver uptake at the intervening zone represents
areas of possible hydrolytic degradation or phase separation
of the silorane adhesive hydrophilic and hydrophobic layers.
Water can compete with the silorane monomer for oxonium
ions, resulting in incomplete polymerization at the adhesive
layer interface [30]. Once stressed by aging or mechanical load-
ing (not tested for our study), this intervening zone potentially
can affect long-term bonding.
Etching dentin with phosphoric acid before to the applica-
tion of silorane self-etch adhesive showed an increase of bond
strength values compared to those of the silorane applied
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according to the manufacturer’s instruction. The literature has
contradictory information regarding phosphoric acid etching
of dentin before the application of one-step self-etch adhe-
sives [31,32]. The efficacy of dentin acid treatment depends
more on the chemical composition of the adhesive than on
the effect of phosphoric acid etching per se [33,34]. Incomplete
resin monomer infiltration at the interface of the hybrid layer
and the undemineralized dentin is a result of a decrease on
gradient of resin monomer diffusion [35] in association with
blocking effect of polyalkenoic acid co-polymer [18]. However,
after aging areas reticular nanoleakage almost vanished from
the bottom of the hybrid layer and the thickness of hybrid
layer significantly reduced. Denuded collagen fibrils within
the hybrid layer become less susceptible to dye penetration
after being aged [36]. Hybrid layer degradation can occur based
not only by hydrolytic degradation but also with host-derived
enzymes [37–39]. When collagen degradation occurs by action
of host-derived matrix metalloproteinases, the result is a par-
tial to complete disappearance of the partially demineralized
collagen matrices [38]. The latter occurrence can explain the
reduction of hybrid layer thickness and nanoleakage at the
bottom of hybrid layer.
The application of a hydrophobic resin coating layer (LS
Bond, 3M ESPE) on top of Single Bond (3M ESPE) signifi-
cantly increased the bond strengths when compared to those
of the SB applied alone. A recent study [28] showed that
it is possible to bond dimethacrylate composite to silorane
composite using a phosphate-methacrylate-based interme-
diate resin. Our study shows that the opposite also is true.
It is possible to bond a silorane composite to a dimethacry-
late based adhesive using a phosphorylated methacrylate (LS
Bond, 3M ESPE) as intermediate resin coating. After aging, the
adhesive layer showed signs of intense silver uptake in com-
parison to uptake after 24 h, though it was not statistically
different from other experimental groups. Water-trees were
observed not only protruding from the hybrid layer towards
the adhesive layer, but also between the adhesive layer and
LS Bond. These water-filled channels are potential sites for
hydrolytic degradation of resin–dentin bonds [19]. In addition,
punctuate nanoleakage became evident within numerous and
small resin globules and blisters at the adhesive layer after
aging. The presence of increased concentrations of dissolved
inorganic ions, uncured, water-penetrable, hydrophilic resin
monomers can trigger a differential water movement across
the cured adhesive layer [25]. Microscopic water droplets
within the blisters can be further replaced by silver uptake
resulting in the spotty nanoleakage. Blister formation in total-
etch adhesive resulting from dentin transudation also was
evidenced in an in vivo experiment [40]. Tay et al. [25] stated
that the blisters also can expand and coalesce with one
another. Therefore, direct clinical application of this bonding
strategy is not recommended until more studies clarify the
bond between methacrylate-based and silorane-based com-
posites.
As an alternative to methacrylate composite for direct
posterior composite restorations, silorane composite resin
represents a new addition to the clinician’s armamentarium.
The main advantage of this new restorative material is its low
shrinkage [41,42]. However, the need of a dedicated adhesive
to achieve acceptable bond strengths restricts the universal
 d e n t a l m a t e r i a l s 2 5 ( 2 0 0 9 ) 589–600
application of silorane. Whereas one-step self-etch adhesive
may be considered as more user-friendly, it still is not able to
fulfill all the requirements for a long-term bonding [10,34,43].
Recent in vivo investigations [10,26,44,45] showed that the
bonding deficiencies of one-step self-etch adhesives still need
to be addressed, in particular bond degradation. A hydropho-
bic resin coating layer may be placed on top of a methacrylate
adhesive to convert silorane into a universal composite, but
further studies are needed to confirm this hypothesis. Hence,
although silorane dedicated adhesive can produce satisfactory
microtensile bond strengths to dentin, nanoleakage was not
completely eliminated.
5. Conclusions
The application of a conventional total-etch methacrylate-
based adhesive followed by a hydrophobic resin coating
produces acceptable bond strengths. Aging does not affect
bond strengths. While pre-etching dentin achieved reliable
bond strengths to silorane dentin adhesive, it does not sig-
nificantly improve the bond strengths to low-shrinkage resin
composite, resulting in no spontaneous debonding. However,
none of the dentin treatments are able to eliminate significant
nanoleakage.
Acknowledgement
The authors thank Dr. Bernard Tandler for editorial assistance.
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