Effects of Multiple Coatings of Two All-in-one Adhesives

on Dentin Bonding
Shuichi Itoa/Franklin R. Tayb/Masanori Hashimotoc/Masahiro Yoshiyamad/Takashi Saitoe/
William W. Brackettf/Jennifer L. Wallerg/David H. Pashleyh

Purpose: Simple changes to bonding technique can improve resin-dentin bond strengths. The objective of this study
was to evaluate the effects of multiple coatings of two all-in-one adhesive resins on both microtensile bond strength
(µTBS) and nanoleakage.

Materials and Methods: The mid-coronal occlusal dentin of extracted human molars was used. Two all-in-one
adhesives – iBond (Heraeus Kulzer) and Xeno III (Caulk Dentsply) – were applied to 320-grit abraded dentin sur-
faces. In groups 1 and 3 during bonding, monomer application and solvent evaporation were done 1, 2, 3, 4 or 5
times on the dentin surface before light curing. In groups 2 and 4 after light curing the first layer, the adhesive was
re-applied, the solvent evaporated, and the layer light cured. This was repeated from 2 to 5 times, followed by
creation of composite buildups. After 24 h storage in 37ºC water, the teeth were sectioned perpendicular to the
adhesive interface to produce multiple beams of composite-bonded dentin, approximately 0.9 mm2 in area. These
were tested to failure in tension. Data were evaluated by three-way ANOVA (material vs coatings vs light curing)
followed by multiple comparisons at α = 0.05. Additionally, nanoleakage of silver uptake and adhesive layer
thickness were evaluated using transmission electron microscopy (TEM).

Results: The results indicated that bond strengths increased with the number of coatings (p < 0.0001) with both
adhesives, up to 3 layers, especially if each layer was light cured. Nanoleakage of silver tended to decrease with
each coat in both adhesive systems.

Conclusion: By simply applying more coats of adhesive, the strength and quality of dentin adhesion can be
improved.

Key words: all-in-one adhesives, dentin bond strength, nanoleakage, multiple coatings.

J Adhes Dent 2005; 7: 133–141. Submitted for publication: 06.02.04; accepted for publication: 05.08.04.

a Instructor, Department of Operative Dentistry and Endodontology, School of
Dentistry, Health Sciences University of Hokkaido, Hokkaido, Japan.
b Assistant Professor, Department of Conservative Dentistry, Faculty of Den-
tistry, The University of Hong Kong, Hong Kong, China.
c Instructor, Division of Pediatric Dentistry, Hokkaido University, Graduate
School of Dental Medicine, Hokkaido, Japan.
d Professor, Department of Operative Dentistry, School of Dentistry, Okayama
University, Okayama, Japan.
e Professor, Department of Operative Dentistry and Endodontology, School of
Dentistry, Health Sciences University of Hokkaido, Hokkaido, Japan.
f Associate Professor, Department of Oral Rehabilitation, School of Dentistry,
Medical College of Georgia, Augusta, Georgia, USA.
g Associate Professor, Division of Biostatistics, Medical College of Georgia,
Augusta, Georgia, USA.
h Regents’ Professor, Department of Oral Biology and Maxillofacial Pathology,
School of Dentistry, Medical College of Georgia, Augusta, Georgia, USA.
Reprint requests: Dr. David H. Pashley, Department of Oral Biology and Maxillo-
facial Pathology, School of Dentistry, Medical College of Georgia, Augusta, GA
30912-1129, USA. Tel: +1-706-721-2033, Fax: +1-706-721-6252. e-mail:
dpashley@mail.mcg.edu
t is well known that bond strength is affected by the
I extent of resin infiltration into the exposed collagen net-
work.3,8 Ideally, adhesive monomers should occupy the
interfibrillar spaces around exposed collagen fibrils.3,14
Dentin adhesives are currently available as three-step,
two-step, and single-step systems, depending on how the
three cardinal steps of etching, priming, and bonding to
tooth substrates are accomplished or simplified.12 Re-
cently introduced all-in-one adhesives further combined

Vol 7, No 2, 2005 133

Ito et al
Table 1 Composition of the all-in-one adhesives used
Material Manufacturer
i-Bond Heraeus Kulzer, Hanau, Germany
SBMP 3M-ESPE, St Paul, MN, USA
Xeno III Dentsply Degussa, Hanau, Germany
A. Universal – 360-02
B. Catalyst – 360-05
Abbreviations: 4-META = 4-methacryloxyethyltrimellitic acid; SBMP = Scotchbond Multi-Purpose Plus adhesive; BHT = 2,6-di-tert-butyl-p-cresol; bis-GMA = (1-methylethylidene)bis[4,1-phe-
nyleneoxy(2-hydroxy-3,1-propanediyl)] bismethacrylate; CQ = camphorquinone (2,3-bornanedione); HEMA = 2-hydroxylethyl methacrylate; PEM-F = Penta-methacryl-oxy-ethyl-cyclo-phos-
phazen-monofluoride; pyro-EMA-SK = tetra-methacryl-ethyl-pyrophosphate; UDMA = urethane dimethacrylate (1,6-dimethacryl-ethyl-oxy-carbonylamino-2,4,4-trimethyl hexane).
these three bonding procedures into a single step. How-
ever, recent studies have shown that some all-in-one
adhesives exhibited relatively low bond strengths when
compared with two-step self-etching systems.2,6,7 As
these products create very thin coatings,6,13 they may be
oxygen inhibited and hence poorly polymerized.19 The
manufacturer’s instructions for iBond recommend appli-
cation of three coats of adhesive prior to evaporation of
solvent and light curing. A recent study also reported that
bonding of Prompt L-Pop may similarly be improved with
an additional layer of adhesive.6,13
Evaluation of silver uptake by transmission electron
microscopy (TEM) can provide clues on the extent of resin
monomer infiltration15 or degree of water uptake.18 Re-
cently, several TEM studies reported incomplete resin in-
filtration into exposed collagen fibrils.10,17 Although there
are two reports showing that multiple coats of Prompt
L-Pop can improve bond strengths to dentin,6,13 this
effect using other all-in-one adhesives has not yet been
determined.
Using the microtensile test, the purpose of this study
was to evaluate the effects on bond strength of multiple
applications of two all-in-one adhesives to dentin with and
without light curing each layer. Additionally, the effects of
multiple coats of all-in-one adhesives on nanoleakage of
the hybrid and adhesive layers were evaluated using TEM.
The null hypotheses tested were that 1 to 5 applications
of all-in-one adhesives have no effect on either bond
strength or nanoleakage, and that there was no difference
between curing each layer vs curing only the last layer.
MATERIALS AND METHODS
Fifty-six extracted noncarious human third molars were
used in this study. The teeth were collected after obtain-
ing the patients’ informed consent under a protocol
approved by the Medical College of Georgia Institutional
Review Board. Flat dentin surfaces were created in
mid-coronal dentin perpendicular to the tooth’s longitudi-
nal axis using a slow-speed diamond saw (Isomet,
134
Composition Application procedure
4-META, UDMA, glutaraldehyde, acetone, Apply three layers with scrubbing, allow
water 30 s of dwell time, evaporate solvent,
light cure for 10 s.
Bis-GMA, HEMA, CQ
A. Universal: HEMA, aerosil R-947 (fumed 1. Dispense liquid from Univeral and
silica), BHT (stabilizer), ethanol, water Catalyst bottles and mix thoroughly.
B. Catalyst: Pyro-EMA-SK, PEM-F, UDMA, 2. Leave on dentin undisturbed for 20 s.
BHT, CQ, p-dimethyl amine ethyl benzoate Gently air dry.
(co-initiator) 3. Light activate for 10 s.
Buehler, Lake Bluff, IL, USA) to remove occlusal enamel
and superficial dentin. Each surface of mid-coronal dentin
was ground with 320-grit silicon carbide paper under
running water for 30 s just prior to bonding.
Experimental Design
Two single-step adhesives were used in this study. The
compositions of these adhesives, Xeno III (Dentsply
Degussa, Hanau, Germany) and iBond (Heraeus Kulzer,
Hanau, Germany) are shown in Table 1.
There were four groups. Group 1 was composed of
Xeno III bonded specimens in which the solvent in each
coating was evaporated but not light cured until the final
coating was applied. Group 2 specimens were bonded
with Xeno III, but the solvent was evaporated from each
layer and the coating was light cured before applying the
next layer. Group 3 specimens were bonded with iBond,
and the solvent was evaporated, but only light cured after
application of the final coat. Group 4 specimens were
bonded with iBond but each layer was both evaporated
and light cured before application of the next layer.
Group 1 (evap)
For Xeno III, equal volumes of liquids A and B were mixed
for approximately 5 s. A generous amount was applied to
the dentin surface. After leaving the adhesive undisturbed
for 20 s, the adhesive was uniformly spread with a gentle
air stream, and the solvent evaporated for 5 s. For
multiple consecutive applications (2, 3, 4, or 5 coats),
adhesive application and solvent evaporation steps were
done repeatedly (Fig 1). In one subgroup, after applying
one coat of A + B liquid and evaporating the solvent,
another coat of liquid B only was applied, the solvent
evaporated and the layer light cured. In another subgroup,
after the first layer was applied and the solvent evaporat-
ed, an additional layer of Scotchbond Multi-Purpose Plus
(3M-ESPE, St Paul, MN, USA) adhesive was applied and
light cured for 10 s.
All adhesives were subsequently light cured for 10 s
with the light-curing unit (Bisco VIP halogen light, Bisco,
Schaumburg, IL, USA) operated at 600 mW/cm2.
The Journal of Adhesive Dentistry

Group 2 (evap + lc)
Each layer of Xeno III was applied, the solvent evaporated
and then the layer was light cured before any further treat-
ment. Thus, each layer was light cured before receiving
the next application.
Group 3 (evap)
For iBond, the bonding resin was applied on the dentin sur-
face and left undisturbed for 30 s. The surface was sub-
sequently gently air dried to evaporate the solvent to form
a slightly shiny adhesive surface. For multiple consecutive
applications of adhesive (2, 3, 4, or 5 coats), the solvent
evaporation steps were done repeatedly but light curing
(10 s) did not occur until the last coat was applied (Fig 1).
Group 4 (evap + lc)
After light curing the first layer, a second layer of iBond
was applied, the solvent evaporated, and the adhesive
light cured for 10 s in a similar manner. These steps were
repeated for 3, 4, and 5 coatings.
Following bonding, five 1 mm increments of TPH resin
composite (Caulk/Dentsply for Xeno III) and Venus (Her-
aeus-Kulzer for iBond) were used to create resin compos-
ite buildups. Each layer was light cured for 40 s.
Statistics
A three-factor ANOVA was used to examine differences in
the mean microtensile bond strength (µTBS). The three
factors were type of adhesive (Xeno III or iBond), light
curing (after each layer or after last layer only), and layers
(1 to 5 layers, 1 layer + B, 1 layer +MP). All two- and
three-factor interactions between the three variables were
included in the model. A Bonferroni correction to the
alpha level was used to determine post-hoc differences.
Statistical significance was assessed using an alpha
level of 0.01, and all statistical analyses were performed
using SAS 8.2 (SAS Institute, Cary, NC, USA).
Microtensile Bond Testing
After the bonded specimens had been stored in water at
37ºC for 24 h, the built-up teeth were sectioned perpen-
dicular to the adhesive interface to remove the peripheral
3 mm from each side of the tooth, yielding a cube-like
specimen that represents the center of the tooth. This
central cube was cut into 0.9-mm-thick slabs. Each slab,
in turn, was sectioned into 1.0-mm-wide beams (adhesive
area: approximately 0.9 mm2) with a diamond saw under
water cooling/lubrication. Although 9 beams were pro-
duced, only the 8 with the greatest dentin thicknesses
were selected for testing. Two teeth were used for each of
the 7 subgroups (14 teeth) per material. Beams prepared
were attached to a testing apparatus with a cyanoacrylate
adhesive (Zapit, Dental Ventures of America, Corona, CA,
USA). A tensile load was applied with a material tester
(Vitrodyne V1000, John Chatillon & Sons, Greensboro,
NC, USA) at a crosshead speed of 1.0 mm/min until
failure, to obtain the ultimate tensile strength, calculated
in MPa. Pretest failures were recorded as zero values in
the calculation of mean bond strengths in each group.
Vol 7, No 2, 2005
Ito et al
Tooth preparation
Bonding resin application These procedures were prepared
1,2,3,4,5 times in groups 1 and 3
Air dry for 5 seconds
with gentle air stream
Light irradiation for 10 seconds These procedures were prepared
1,2,3,4,5 times in groups 2 and 4
Incremental resin composite buildup and
light irradiation of each layer for 60 sec.
Fig 1 Bonding, evaporation, light-curing steps in the experimental
groups.
Transmission Electron Microscopy (TEM) of Nanoleakage
Four additional teeth with flat dentin surfaces produced in
the same manner as for microtensile bond testing were
employed for each of the subgroups in Groups 1 to 4
(Table 1). After treating the dentin surfaces with the
respective bonding protocol, a microfilled resin compos-
ite containing pre-polymerized fillers (EPIC-TMPT, Parkell,
Farmingdale, NY, USA) was coupled to the bonded dentin
in order to facilitate ultramicrotomy. The bonded speci-
mens were stored in distilled water at 37°C for 24 h. A
1-mm-thick slab was sectioned from each specimen using
the slow-speed saw under water cooling. The slabs were
immersed in 50 wt% ammoniacal silver nitrate for 24 h,
according to a TEM silver tracer technique.16 Following
the reduction of the diamine silver ions into metallic silver
grains, the slabs were dehydrated in an ascending etha-
nol series and embedded in epoxy resin in preparation for
ultramicrotomy. Ninety- to 100-nm-thick undemineralized
sections containing the resin-dentin interfaces were col-
lected on single-slot, carbon- and formvar-coated copper
grids. They were examined without further staining using
a transmission electron microscope (Philips EM208S,
Eindhoven, The Netherlands) operating at 80 kV.
RESULTS
Effects on Bond Strength of Multiple Coats Without
Curing each Layer
Groups 1 and 3: Xeno III (evap) and iBond (evap)
Single applications of Xeno III or iBond produced low µTBS
(7.2 ± 6.3 and 12.2 ± 7.5 MPa, respectively) that were
not statistically different. Four of the 16 beams bonded
with one coat of Xeno III in group 1-1 (evap) exhibited pre-
test failures (Table 2). No other subgroups in groups 1 and
3 showed any pretest failures. The second layer of Xeno
III increased the bond strength, although the increase was
not statistically significant (Table 2). The effect of multiple
applications of Xeno III reached significance after three
135
Ito et al

Table 2 Effects of multiple XIII and iBond adhesive coatings on microtensile bond strengths to dentin

Layers
Groups 1 2 3 4 5 1+B 1 + SBMP

1 X (evap) 7.2 ± 6.3a 22.6 ± 9.2a,b 30.0 ± 9.9b 43.5 ± 7.7c 41.4 ± 8.0c 28.4 ± 9.3b 27.8 ± 5.9b
4/16 0/16 0/16 0/16 0/16 0/16 0/16

2 X (evap + lc) 7.2 ± 6.3a 47.3 ± 19.2b 65.4 ± 20.4d 81.8 ± 20.8d 68.9 ± 22.6d 24.4 ± 15.2b 64.7 ± 21.2d
4/16 0/16 0/16 0/16 0/16 0/16 0/16

3 iB (evap) 12.2 ± 7.5A 18.5 ± 6.2A 30.6 ± 7.0b 34.2 ± 6.0b,c 51.6 ± 14.8b,c – 62.3 ± 23.0c
0/16 0/16 0/16 0/16 0/16 0/16

4 iB (evap + lc) 12.2 ± 7.5A 39.6 ± 15.2B 42.7 ± 12.3b,c 36.6 ± 7.7b 35.8 ± 13.4b – 58.0 ± 23.5c
0/16 0/16 0/16 0/16 0/16 0/16

Values are mean ± SD (n = 16) MPa. In groups 1 and 2, 4 out of the 16 failed prematurely. Zero values were used for those 4 specimens in each group. (evap) = solvent evaporation
between coatings but no light curing; (evap + lc) = solvent evaporation and light curing between each coating. Superscript letters designate statistically different (p < 0.01) groups.
1 + B = after applying the first coating, a second coating of solution B was applied. 1 + MP = after applying the first layer, an additional layer of Scotchbond Multi-Purpose Plus adhesive
was applied and light cured.

layers (p < 0.001) and a plateau after four coats, since
there was no statistically significant difference between
four and five coats (Table 2). When one coat of Xeno III
was covered with a second layer of solution B and then
light cured, the µTBS was not different than if a second
layer of A + B had been applied (Table 2). Similarly, when
one coat of Xeno III was covered with one layer of Scotch-
bond Multi-Purpose Plus (SMBP) adhesive and then light
cured, there was no significant improvement in bond
strength over using a second coat of Xeno III.
Using iBond (Group 3), application of two coats did not
significantly increase µTBS, although three coats (the
manufacturer’s instruction) significantly (p < 0.002) in-
creased bond strength (Table 2). There was no significant
effect of a fourth or fifth coat on improving bond strength.
Covering one coat of light-cured iBond with SBMP adhe-
sive significantly improved bond strength (p < 0.002) by a
factor of five as compared to a one-coat application.
Using SBMP adhesive increased the µTBS 3.4 times
(p < 0.002) over that produced by a second coating of
iBond (Table 2).
Effects on Bond Strength of Individually-cured Multiple
Coats
Groups 2 and 4: Xeno III (evap + lc) and
iBond (evap + lc)
When a second layer of adhesive was applied to the al-
ready light-cured first layer in Group 2 specimens bonded
with Xeno III, the microtensile bond strength increased
6.6-fold (p < 0.002) over the single coat value (compare
7.2 ± 6.3 to 47.3 ± 19.2, Table 2). Adding a third coating
significantly (p < 0.002) increased bond strength again to
65.4 MPa. Addition of a fourth and fifth coat did not
significantly increase bond strength over that produced by
three coats. When one coat of light-cured Xeno III was
covered with another coat of bottle B, there was no
significant increase in bond strength over that produced
by one coat of Xeno III alone. However, when one coat of
light-cured Xeno III was covered with one coat of
light-cured SBMP adhesive, the bond strengths increased
significantly to 64.7 MPa.
When the second layer of adhesive was applied to the
already light-cured first layer in Group 4 specimens
bonded with iBond, the bond strength increased and was
as high as three layers of iBond where only the last layer
was light cured (Table 2). Addition of a third, fourth, or
fifth light-cured layers produced no further increase in
bond strength. When one layer of light-cured iBond was
covered with one layer of light-cured SBMP adhesive, the
bond strength was significantly (p < 0.002) higher than a
second, third, fourth, and fifth coat of light-cured iBond.
Effects on Nanoleakage of Multiple Coatings not Cured
Individually
TEM of specimens treated with one layer of iBond (not
used according to the manufacturer’s directions of apply-
ing three layers) revealed direct contact of the resin
composite with the surface of dentin (Fig 2a). That is,
there was no layer of polymerized adhesive between the
top of the hybrid layer and the bottom of the composite.
Nanoleakage in the form of heavy silver deposits could be
seen within the 1-µm-thick, partially demineralized hybrid
layer (Fig 2a). When one layer of Xeno III was used, the
hybrid layer was covered by 2 to 3 µm of adhesive
(Fig 2b). The amount of nanoleakage was similar when a
second coat of adhesive was applied before light curing,
which created a 4-µm-thick adhesive layer above a
1-µm-thick hybrid layer (not shown). Both all-in-one adhe-
sives removed most of the smear layer, although some
smear plugs remained in some tubule orifices.
With the application of three consecutive coats of
iBond prior to light curing, the thickness of the adhesive
layer increased to 7 to 8 µm (Fig 2c). The increase in
thickness of the adhesive layer was also accompanied by
an increase in the propensity to form linear and spherical

136 The Journal of Adhesive Dentistry


Fig 2a TEM micrographs of specimens bonded with one coat of
iBond that had the solvent evaporated prior to application of the next
layer but no light-curing until application of the final layer. All speci-
mens were undemineralized and were immersed in AgNO3 prior to
embedding to reveal nanoleakage. C: composite; D: intertubular
dentin; P: peritubular dentin. One-coat of iBond showing direct
contact of composite with the dentin surface, with an absence of an
adhesive layer. Silver deposits (arrow) were seen within the 1 µm
thick hybrid layer and between the dentin and composite and within
blisters (pointers).
Fig 2c TEM micrographs of specimens bonded with three coats of
iBond that had the solvent evaporated prior to application of the next
layer but no light-curing until application of the final layer. All speci-
mens were undemineralized and were immersed in AgNO3 prior to
embedding to reveal nanoleakage. C: composite; D: intertubular
dentin; P: peritubular dentin. Sporadic silver uptake was seen in the
1-2 µm thick hybrid layer (H). In the adjacent 7-8 µm thick adhesive
layer (A), large accumulations of silver were seen (2-3 µm in diame-
ter globular structures adjacent to the hybrid layer with occasional
1 x 8-10 µm linear deposits of silver extending across the adhesive
layer).
silver-filled structures within the iBond adhesive layer
(Fig 2c). In addition, numerous isolated silver grains
could be identified throughout the entire adhesive layer
(Fig 2c). Extensive nanoleakage could be seen within the
hybrid layer. Three coats of Xeno III exhibited less silver
uptake into the hybrid layer compared to three coats of
Vol 7, No 2, 2005
Ito et al
Fig 2b TEM micrographs of specimens bonded with one coat of
Xeno III that had the solvent evaporated prior to application of the
next layer but no light-curing until application of the final layer. All
specimens were undemineralized and were immersed in AgNO3 prior
to embedding to reveal nanoleakage. C: composite; D: intertubular
dentin; P: peritubular dentin. An adhesive (A) layer 2 µm thick is
seen over a 2 µm thick hybrid layer (between open arrows). Silver
uptake was seen sporadically within the hybrid layer, surrounded by
large areas of hybrid layer free of silver.
Fig 2d TEM micrographs of specimens bonded with three coats of
Xeno III that had the solvent evaporated prior to application of the
next layer but no light-curing until application of the final layer. All
specimens were undemineralized and were immersed in AgNO3 prior
to embedding to reveal nanoleakage. C: composite; D: intertubular
dentin; P: peritubular dentin. Three-coats of Xeno III produced a
2-3 µm thick hybrid (H) layer that showed scattered silver uptake and
a relatively thick 8-9 µm layer of silica filled adhesive (A) that
exhibited only individual spot-like silver uptake.
iBond (compare Figs 2c vs 2d), although the Xeno III
adhesive showed similar isolated spot-like silver grains.
A further increase in the number of adhesive coats to
five layers did not necessarily result in an overall increase
in the thickness of the hybrid layer or the polymerized
adhesive layers (Figs 2e and 2f), but it did decrease the
137
Ito et al
Fig 2e TEM micrographs of specimen bonded with five coats of
iBond that had the solvent evaporated prior to application of the next
layer but no light-curing until application of the final layer. All speci-
mens were undemineralized and were immersed in AgNO3 prior to
embedding to reveal nanoleakage. C: composite; D: intertubular
dentin; P: peritubular dentin. Five coats of iBond produced a 1-2 µm
thick hybrid layer (H) with only sporatic silver uptake. The adhesive
layer (A) was only 7-8 µm thick but was almost free of silver
nanoleakage that was limited to spot-like deposits (open arrow-
heads). Silver deposits were seen both in the bottom of the hybrid
layer (white arrow) and in the top of the hybrid layer (pointer).
amount of nanoleakage in the hybrid layers produced by
both adhesives.
Effect on Nanoleakage of Multiple Coatings Cured
Individually
Figure 3a is an overview of the resin-dentin interface that
was bonded using three coats of iBond that were light
cured separately, creating an adhesive layer that was
about 17 to 20 µm thick. The amount of nanoleakage in
the adhesive layer was less than that observed in the
comparable 3-coat group, where the layers were not light
cured (compare Fig 2c vs Fig 3a). Although the thickness
of the adhesive layer was similar to that achieved previ-
ously with the use of three consecutive coats of adhesive
(Fig 2c), there was a complete absence of linear distribu-
tions of silver within with the adhesive when these coats
were light cured separately (Fig 3a). This did not, however,
eliminate the heavy silver staining within the hybrid layer.
The absence of linear silver-filled channels in the adhe-
sive of iBond (Fig 3a) and the gradual reduction in the size
of the isolated silver grains (Fig 3c) became more promi-
nent with the use of five coats of separately light-cured
iBond adhesives, which generated a 35-µm-thick adhe-
sive layer between the composite and dentin, although
nanoleakage in the hybrid layer was severe. Similar
results were obtained using three vs five coats of Xeno III
(Figs 3b and 3d), although the amount of nanoleakage in
the hybrid layer was much less than was seen in hybrid
layers created by iBond.
The difference between the use of multiple coatings of
iBond or Xeno III and the use of the Scotchbond Multi-Pur-
pose resin over a single layer of these cured adhesives is
138
Fig 2f TEM micrographs of specimen bonded with five coats of
Xeno III that had the solvent evaporated prior to application of the
next layer but no light-curing until application of the final layer. All
specimens were undemineralized and were immersed in AgNO3 prior
to embedding to reveal nanoleakage. C: composite; D: intertubular
dentin. Five coats of Xeno III produced a 1 µm thick hybrid layer
(H between two open arrows) that showed minimal silver uptake,
and a 10 µm thick adhesive layer (A) that showed only spot-like
silver deposits.
shown in Figs 3d and 3f. Although the thickness of the hy-
brid layers and the nanoleakage within the hybrid layer
were not very different from those previously described,
there was a complete absence of silver uptake within the
top of the 15-µm-thick adhesive layer (Figs 3d and 3f). The
most distinctive features, however, were the dramatic re-
duction in the size of the isolated silver grains within the
intermixed zone between the all-in-one adhesives and the
Scotchbond Multi-Purpose resin (Fig 3e), followed by the
total elimination of these silver grains in the latter (Fig 3f).
DISCUSSION
The results of this study require rejection of the null
hypothesis that multiple coatings of these two all-in-one
adhesives do not increase µTBS. The results also require
rejection of the second null hypothesis that there is no
difference in µTBS regardless of whether the multiple
layers were light cured.
The TEM observations indicate that one layer of iBond
(not used according to the manufacturer’s recommenda-
tion) is insufficient to adequately create a hybrid layer
covered by an adhesive layer that will provide sufficient
coupling to subsequently applied resin composite. How-
ever, one coating of Xeno III which contains nanofillers
was enough to create a hybrid layer covered by an adhe-
sive layer. Although two or more coatings with either
material dramatically increased µTBS when both layers
were light cured, there was a further significant increase
in µTBS (Table 2). This was especially evident using Xeno
III. Using three layers of Xeno III without light curing
The Journal of Adhesive Dentistry

Fig 3a TEM micrographs of specimen bonded with three coats of
iBond that had the solvent evaporated from each layer and in which
each layer was light-cured before application of the next layer. All
specimens were undemineralized and had been immersed in AgNO3
to reveal nanoleakage. One-coat specimens are not shown since
they were the same as Figs 2a and 2b. Three-coats of iBond
produced a 2 µm thick hybrid layer (H) and a 17-20 µm thick adhe-
sive layer. Although there was considerable silver uptake by the
hybrid layer, the overlying adhesive (A) layer was devoid of major
silver uptake, showing only tiny silver grains scattered throughout
the layer. C: composite; D: intertubular dentin.
Fig 3c TEM micrographs of specimen bonded with five coats of
iBond that had the solvent evaporated from each layer and in which
each layer was light-cured before application of the next layer. All
specimens were undemineralized and had been immersed in AgNO3
to reveal nanoleakage. One-coat specimens are not shown since
they were the same as Figs 2a and 2b. Five-coats of iBond produced
an adhesive (A) layer 34 µm thick that was free of silver uptake,
although the hybrid layer (H) showed severe silver nanoleakage.
C: composite; D: intertubular dentin.
between layers (but with solvent evaporation at each
step) produced a mean µTBS of 30.0 MPa. However, if
each layer was light cured, three layers gave a µTBS of
65.4 MPa (p < 0.05 compared to three coats, of which
only the last one was light cured).
iBond contains water and acetone as solvents. The
evaporation of acetone is much faster than that of water.
After acetone is gone, water must evaporate by itself,
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Ito et al
Fig 3b TEM micrographs of specimen bonded with three coats of
Xeno III that had the solvent evaporated from each layer and in which
each layer was light-cured before application of the next layer. All
specimens were undemineralized and had been immersed in AgNO3
to reveal nanoleakage. One-coat specimens are not shown since
they were the same as Figs 2a and 2b. Three-coats of Xeno III
produced a 2 µm thick hybrid layer (H) and a 12-13 µm thick adhe-
sive (A) layer that contained little silver. C: composite; D: intertubular
dentin.
Fig 3d TEM micrographs of specimen bonded with five coats of
Xeno III that had the solvent evaporated from each layer and in which
each layer was light-cured before application of the next layer. All
specimens were undemineralized and had been immersed in AgNO3
to reveal nanoleakage. One-coat specimens are not shown since
they were the same as Figs 2a and 2b. Five-coats of Xeno III
produced a 22-24 µm thick adhesive (A) layer that took up little
silver. The hybrid layer (H) was almost free of silver. C: composite;
D: intertubular dentin.
albeit at a much lower vapor pressure. Thus, multiple
applications may tend to permit the accumulation of
water on the surface. Such water should be evaporated to
avoid low bond strengths.11
Solution B in Xeno III contains all of the comonomers,
initiators, and accelerators except HEMA (Table 1). It
contains no solvent. Solution A contains HEMA and a
water-ethanol mixture. It was thought that adding an
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Ito et al
Fig 3e TEM micrographs of specimen bonded with one coat of
iBond that had the solvent evaporated and then was light-cured
before application of Scotchbond Multi-Purpose (SBMP) adhesive
that was subsequently light-cured. All specimens were undemineral-
ized and had been immersed in AgNO3 to reveal nanoleakage.
One-coat specimens are not shown since they were the same as
Figs 2a and 2b. One layer of iBond (light-cured) followed by the
application of one layer of Scotchbond Multi-Purpose (SBMP) adhe-
sive to dentin. This treatment produced a 15 µm thick adhesive layer
that consisted of 9 µm of SBMP (solid line R) and 8 µm of iBond
adhesive (dotted line A) with a 2 µm thick intermixed zone (IZ). There
was much less silver uptake into the hybrid layer created by a single
coat of iBond compared to its control (Fig 2a). C: composite;
D: intertubular dentin; H: hybrid layer.
additional coating of B (without A) to the first coating of
A + B would enhance the µTBS of Xeno III because it
would be less hydrophilic. This was unsuccessful
(Table 2), regardless of whether the first layer was light
cured or not prior to the application of material from
solution B (that was subsequently light cured).
When one layer of iBond or Xeno III was covered with
SBMP adhesive, there was a significant increase
(p < 0.05) in µTBS (Table 2) regardless of whether the
first coat was light cured or not. The adhesive of SBMP is
basically a blend of bis-GMA/HEMA without any volatile
solvent. This adhesive layer may have increased the
mechanical properties of the bond due to improved
polymerization. The more hydrophobic SBMP adhesive
may have also sealed the surface of the more hydrophilic
iBond. Indeed, Figs 3e and 3f shows that almost no silver
uptake was seen in the SBMP adhesive layer (R), unlike
the substantial silver uptake seen in the underlying iBond
and Xeno III adhesives.
It is likely that the increased bond strength seen in
both all-in-one adhesives with multiple coatings is due to
several mechanisms operating simultaneously. As the
first layer of adhesive begins to etch dentin, it is probably
rapidly buffered,9 so that additional layers of unpolymer-
ized comonomers may improve the etching ability of
adhesives. Although the hybrid layer thickness of iBond
specimens increased from 0.8 to 1.5 µm when one vs
three layers was applied, the values were not statistically
significantly different. When using multiple coatings of
140
Fig 3f TEM micrographs of specimen bonded with one coat of
Xeno III that had the solvent evaporated from each layer and then
was light-cured before application of Scotchbond Multi-Purpose
(SBMP) adhesive that was subsequently light-cured. All specimens
were undemineralized and had been immersed in AgNO3 to reveal
nanoleakage. One-coat specimens are not shown since they were
the same as Figs 2a and 2b. One layer of Xeno III (evaporated and
light-cured) followed by the application of one layer of SBMP to
dentin produced a 14-15 µm thick adhesive layer that consisted of
12-13 µm of SBMP (line R) and a 5 µm thick layer of Xeno III (A) with
a 2-3 µm thick intermixed zone (IZ). Although there was moderate
silver uptake by the hybrid layer (H), there was no silver uptake in the
adhesive layer (A). C: composite; D: intertubular dentin.
adhesives, there are several techniques that can be
employed. If the solvent is evaporated between each
coat, the concentration of comonomers that exists after
each coating should increase, thereby facilitating
comonomer infiltration with minimal increase in the
thickness of the adhesive layer.
These solvated adhesives only contain 25 to 30 wt%
comonomers and 70 to 85% solvents (preliminary gravi-
metric analysis). The presence of the solvent facilitates
wetting and spreading of the solvated comonomers and
may assist in preserving the width of interfibrillar spaces
during infiltration.3,5 This may facilitate inward diffusion
of the comonomers into the matrix. When most of the
solvent is evaporated, the concentration of the adhesives
must increase dramatically, although the thickness of the
layer may be extremely thin.6,13 The cumulative concen-
tration of these constituents must rise in the adhesive
layer with each application. We speculate that this con-
tributes to improved polymerization of the final mixture.19
Although the thickness of the resin layer increases
enough to prevent oxygen inhibition of its entire thick-
ness,6,13 we believe that the final adhesive layer has an
improved quality (ie, better cross linking, improved inter-
penetration of polymer chains, improved percent conver-
sion). We speculate that the bond strength reached a
plateau following sequential coatings when a limiting
critical variable had been reached (eg, film thickness,
cross-linking density, concentration of photoinitiators,
accelerators, etc).
The Journal of Adhesive Dentistry

In contrast, when each layer was light cured following
solvent evaporation, the improved bond strengths may
have been due to mechanical rather than chemical
factors. After the first layer was cured, it would be unlikely
that there could be any more resin infiltration into the ma-
trix. However, it might be possible to improve the quality
of each layer of adhesive. That is, since each layer was
polymerized, only the oxygen-inhibited portion of the layer
would be available to mix with the newly applied layer.
However, the oxygen-inhibited layer in iBond can be 8 to
10 µm thick and can easily mix with subsequently applied
layers. This might improve its quality. One layer of iBond
produced an adhesive layer thickness that ranged from
0 to 3 µm. When three layers were applied and each one
was light cured, the adhesive layer was about 15 to
17 µm thick. Five layers of iBond (each cured individually)
produced a 34-µm-thick adhesive layer (Fig 3c), while
five layers of individually cured Xeno III produced a
26-µm-thick adhesive layer. We speculate that the major
improvement in bond strength with successive layers in
group 2 and 4 specimens was due to increased adhesive
layer thickness.4 Multiple layers of adhesive tend to cre-
ate thicker adhesive layers that increase the compliance
of the joint. This may lower local stress concentrations
during tensile testing, resulting in higher yield strengths.
Regardless of the mechanism(s) responsible for
increased bond strength, the need for multiple applica-
tions increases the bonding time of these 'simplified'
adhesives. Although we did not attempt to optimize the
solvent evaporation time, we used 5 s. Five seconds for
each of 4 layers adds 20 s to the procedure. However,
adding these 20 s, plus light curing each of the four
layers for 10 s (40 s cumulative curing time plus 20 s
evaporation time), requires more time than is stated
current manufacturer’s instructions.
These results confirm how important bonding tech-
nique is to producing optimal resin-dentin bonds.1,20
Simple changes in bonding technique, such as applying
more layers of all-in-one adhesives, can lead to large
increases in initial bond strength. Whether this improves
the long-term durability of resin-dentin bonds remains to
be determined.
ACKNOWLEDGMENTS
We thank Amy Wong of the Electron Microscopy Unit, The University of Hong
Kong for technical assistance and Michelle Barnes of the Medical College of
Georgia for secretarial support. This work was supported, in part, by grants DE
014911 and DE 015306 (to DHP) from the National Institute of Dental and
Craniofacial Research. The materials (iBond/Heraeus Kulzer, Hanau, Germany
and Xeno III/Caulk Dentsply, Germany) used in this study were generously
supplied by each manufacturer. The testing device used for microtensile bond
testing (Ciucchi’s jig) was supplied by Kuraray Co, Ltd, Osaka, Japan.
Vol 7, No 2, 2005
Ito et al
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Clinical relevance: By simply applying more coats of
adhesive, the strength and quality of dentin adhesion
can be improved.
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