Aust Endod J 2010

ORIGINAL RESEARCH

Inuence of endodontic irrigants on bond strength of a

self-etching adhesive
aej_249

1..5

Ana P. Farina, DDS, MSc1; Doglas Cecchin, DDS, MSc1; Joo V.B. Barbizam, DDS, MSc, PhD2; and
Bruno Carlini-Jnior, DDS, MSc, PhD2
1 Department of Restorative Dentistry, School of Dentistry of Piracicaba, State University of Campinas (Unicamp), Piracicaba, So Paulo, Brazil
2 Department of Restorative Dentistry, School of Dentistry of Passo Fundo, University of Passo Fundo (UPF), Passo Fundo, RS, Brazil

Keywords
chlorhexidine, dentine-bonding agents, EDTA,
microtensile, sodium hypochlorite.
Correspondence
Ms Doglas Cecchin, Department of Restorative
Dentistry, Piracicaba Dental School, State
University of Campinas, Avenue Limeira 901,
Caixa Postal 52, Piracicaba, So Paulo, Brazil.
Email: dgscecchin@yahoo.com.br
doi:10.1111/j.1747-4477.2010.00249.x

Abstract
The aim of this in vitro study was to evaluate the bond strength of a self-etching
adhesive system to dentine irrigated with sodium hypochlorite (NaOCl), chlorhexidine solution (CX) and ethylene diamine tetra acetic acid (EDTA) by
microtensile testing. Sixty human third molars were sectioned 3 mm below the
occlusal level and then randomly divided into six groups: G1(control), without
irrigant solution; G2, 1% NaOCl; G3, 1% NaOCl followed by the application of
17% EDTA; G4, 2% CX; G5, 2% CX followed by the application of 17% EDTA
and G6, 17% EDTA. The specimens received the self-etching adhesive system
were restored with composite resin, then sectioned and trimmed to obtain four
hourglass-shaped slabs from each tooth. The specimens were submitted to the
microtensile test in a Universal Testing Machine at a crosshead speed of
0.5 mm min-1 until fracture. The results were submitted to statistical analysis
by ANOVA/Newman-Keuls. The means and standard deviations (MPa) were:
G1, 26.88 (3.81); G2, 19.08 (3.89); G3, 18.16 (2.21); G4, 18.14 (4.32);
G5, 34.30 (3.32); G6, 13.61 (1.21). It was concluded that the application of
2% CX followed by the application of 17% EDTA resulted in increasing the
bond strength of the self-etching adhesive system to dentine, when compared
with the results obtained for the other tested groups.

Introduction
Endodontically treated teeth generally lose tooth structure as a result of trauma, caries and endodontic procedures (1) that change their physical properties and
compromise their aesthetic appearance (2). The purpose
of restoring endodontically treated teeth is to prevent
bacterial infiltration from the oral cavity, re-establish
their functionality and aesthetics and avoid fracture of
the remaining dental structure (3). Vire in 1991 (4) verified that 59.4% of the failures in teeth submitted to root
canal treatment occur during re-establishment of the lost
dental structure.
Chemical substances used during biomechanical preparation of root canals can alter the composition of the
dentine surface and affect the interaction with restorative
materials (5,6). Sodium hypochlorite (NaOCl) and ethyl-

ene diamine tetra acetic acid (EDTA) are substances

usually used during endodontic treatment (7). NaOCl is
an auxiliary irrigant solution used in the instrumentation
process and it results in debridement, lubrication, disinfection, tissue dissolution, collagen layer removal and
dentine dehydration (8,9). EDTA is indicated as a final
irrigating agent, for the purpose of demineralising the
dentine and promoting appropriate cleaning of the root
canal walls, thus improving the penetration of chemical
substances and promoting a better contact between the
dentine walls and filling material (10). Chlorhexidine has
been shown to be as effective as NaOCl against microorganisms and has therefore been used as an auxiliary solution to instrumentation during endodontic treatment
(11,12).
Endodontically treated teeth with a sufficient amount
of sound coronal structure should preferably be restored

Published 2010. This article is a US Government work and is in the public domain in the USA.

Endodontic Irrigants on Bond Strength

A. P. Farina et al.

with composite resin by the direct technique, because of

its capacity to bond to dentine and increase the fracture
resistance of the remaining dental structure (6,1315).
This process requires appropriate interaction of the adhesive system with the dentine substrate (16). However, the
irrigating substances frequently used during endodontic
treatment could interfere in the bond strength of the
composite bond to dentine (5,6).
Therefore, the aim of the present in vitro study was to
evaluate the influence of endodontic irrigating solutions
on the microtensile bond strength of a self-etching adhesive system to dentine.

Materials and methods

Experimental design
The factor under study was the irrigating solution: G1
without irrigating solution (control), G2 1% NaOCl
(Natufarma pharmacy; Passo Fundo, RS, Brazil); G3 1%
NaOCl followed by the application of 17% EDTA (Biodynamics; Ibipor, PR, Brazil); G4 2% chlorhexidine solution (CX) (Farmcia Natufarma, Passo Fundo, RS, Brazil);
G5 2% CX followed by the application of 17% EDTA;
G6 17% EDTA. The restorative system was Clearfil SE
Bond self-etching adhesive (Kuraray; Okayama, Japan)
and Z250 composite resin (3M ESPE; St Paul, MN, USA).
The experimental units consisted of slabs of human
dentine randomly distributed into the six experimental
groups (n = 40). The response variable was microtensile
bond strength evaluated in MPa.

Selection of teeth
Sixty sound freshly extracted human third molars were
used in this study. Teeth were stored 0.5% chloramine
solution at 4C for 48 h for disinfection. Next, the teeth
were cleaned with pumice slurry/water using Robinson
brushes (Microdont; Socorro, SP, Brazil) and analysed
under 10 magnifying glass (Carl Zeiss, Jena, Germany)
to exclude those with fracture lines or fissures. The teeth
were stored in distilled water at 4C.

Microtensile test
Each tooth was individually embedded in a PVC cylinder
(25 mm diameter and 20 mm height) (Tigre; So Paulo,
SP, Brazil) using colourless autopolymerising acrylic resin
(Jet Clssico; So Paulo, SP, Brazil), so that the occlusal
surface faced upwards. The teeth were sectioned 3 mm
below the occlusal surface in a metallographic sectioning
machine (Struers Minitom; Copenhagen, Denmark) and
were polished (Struers Abramin; Copenhagen, Denmark)
2

with successively finer grades of silicon carbide papers

(#600 to 1200-grit). The samples were washed for 60 s
and stored in distilled water at 4C for 24 h.
The specimens were randomly distributed into the
experimental groups. In group 1 (control), no irrigating
solution was applied. In group 2, 5 mL of 1% NaOCl was
applied to the surface for 40 min*. NaOCl solution (5 mL)
was reapplied on the dentin every 5 min. In group 3, 1%
NaOCl was applied following the protocol of group 2, in
addition to an application of 17% EDTA for 5 min*. In
group 4, 5 mL of 2% CX was applied to the surface for
40 min*. CX solution (5 mL) was reapplied on the
dentine every 5 min. In group 5, 2% CX was applied
following the protocol of group 4 in addition to an application of 17% EDTA for 5 min*. In group 6, 17% EDTA
(5 mL) was applied to the surface for 5 min*. After this
period, the specimens were washed with distilled water
for 60 s. (*This corresponds to the clinical time that
irrigating substances are used in root canals during
endodontic treatment.)
In order to perform the adhesive technique in accordance with the manufacturers instructions, the dentine
received two layers of Clearfil SE Bond self-etching adhesive system using microbrush tips (KG Sorensen, Barueri,
SP, Brazil) followed by light polymerisation for 20 s (Radii
device; SDI, So Paulo, SP, Brazil) at 450 Mw cm-2 of light
intensity.
After hybridisation, three increments (~2 mm each) of
Z250 composite resin (Shade A2) were applied on the
dentine surface with a #1/2 spatula, reaching a total height
of 6 mm. Each increment was light polymerised for 20 s
with the Radii device. After this, the specimens were
stored in distilled water for 24 h.
The samples were placed in a metallographic sectioning
machine (Struers Miniton, Copenhagen, Denmark) and a
diamond double face disk, under water cooling, was used
to cut sequential 1.0 mm thick sections (in the mesiodistal direction) to the long axis of the specimens, carefully so as not to separate the slices. The specimens were
then separated from the acrylic resin base through a
transversal section, to obtain resin/dentine slabs measuring approximately 10 mm high, 5 mm wide and 1 mm
thick. The specimens were trimmed on both sides of
resindentine interface using a #1093 FF drill (KG
Sorensen, Barueri, SP, Brazil) at high speed (Kavo, Joinvile, SC, Brazil). The purpose of this procedure was to
obtain a central area 1 mm thick, and configure standard
hourglass-shaped samples (Fig. 1).
The samples were individually fixed in a metal device
with a cyanoacrylate adhesive (Loctite Super Bonder, So
Paulo, SP, Brazil) so that the resin/dentine interface
remained without any contact, allowing the microtensile
test. The metal device was placed in a Universal Testing

Published 2010. This article is a US Government work and is in the public domain in the USA.

Endodontic Irrigants on Bond Strength

A. P. Farina et al.

Table 1 Mean microtensile bond strengths (MPa) of coronal dentine for

different irrigation solutions

Figure 1 Schematic of specimen preparation. (1) exposed dentine, (2)

restored tooth, (3) sectioning of the tooth to obtain four slabs of 1 mm
thick per each tooth and (4) specimen with hourglass shape.

Groups

Irrigating solution

Mean
values

Standard
deviations

G1
G2
G3
G4
G5
G6

Without irrigating solution

1% NaOCl
1% NaOCl + EDTA 17%
2% CX
2% CX + EDTA 17%
EDTA 17%

26.88
19.08
18.16
18.14
34.30
13.61

3.81b
3.89c
2.21c
4.32c
3.32a
1.21d

Same letters indicated statistical similarity (a = 0.01). CX, chlorhexidine

solution; EDTA, ethylene diamine tetra acetic acid; NaOCl, sodium
hypochlorite.

Table 2 Modes of bond failure (%)

Failure mode

Machine (Emic DL 2000, So Jos de Pinhais, PR, Brazil)

and the specimens subjected to the microtensile test at a
crosshead speed of 0.5 mm min-1 until fracture. At the
moment of fracture, the resistance values were recorded
in Newtons (N) by computer software.
Before the test, the area was measured with a digital
pachymeter (Vonder Digital Electronic Paqumetro,
Curitiba, PR, Brazil) and the bond strength was calculated
in MPa by the following formula: Rt = F/A, where Rt is the
microtensile bond strength, F is the force applied and A is
the bond area between the dentine and restorative
system. The data obtained were submitted to ANOVA and
Newman Keuls tests (a = 0.01).
After the microtensile test, the two parts of the specimens were examined under a microscope D. F. Vasconcellos (So Paulo, SP, Brazil) at 40 magnification to
define the failure modes. They were classified into one or
four types: type A, interfacial failure, located entirely
between the adhesive and dentine; type B, mixed failure,
if the fracture site continued from the adhesive into either
the resin composite or dentine; type C, cohesive failure in
dentine; and type D, cohesive failure in resin composite.
The hypothesis tested in this study was that endodontic
irrigating substances (NaOCl, CX and EDTA) adversely
affect the bond strength of the Clearfil SE Bond selfetching adhesive system to dentine, for all groups under
study.

Groups

Irrigating solution

CD

CR

G1
G2
G3
G4
G5
G6

Without irrigating solution

1% NaOCl
1% NaOCl + EDTA 17%
2% CX
2% CX + EDTA 17%
EDTA 17%

20
73
73
29
14
72

70
27
23
29
19
21

6
0
4
40
60
7

4
0
0
2
7
0

Failure mode: A, interfacial failure; CD, cohesive failure in dentine; CR,

cohesive failure in resin composite; M, mixed failure. CX, chlorhexidine
solution; EDTA, ethylene diamine tetra acetic acid; NaOCl, sodium
hypochlorite.

to dentine among the tested groups, including the control

group. The groups treated with 1% NaOCl, 1% NaOCl
followed by 17% EDTA and 2% CX solution had a significantly lower bond strength than the group treated
with 2% CX followed by 17% EDTA and the control
group, but had a significantly higher bond strength than
the Group treated with 17% EDTA.
The failure mode analysis percentages are shown in
Table 2. The analysis showed that the failures were adhesive and mixed in most of the groups, except for the
group that received treatment with CX and the group
with CX followed by EDTA, in which the largest number
of failures was cohesive in dentin. The cohesive failures in
resin were low and not significant.

Results

Discussion

Table 1 shows the mean values and standard deviations

(MPa) of groups tested in this study.
There was significant statistical difference for irrigating
solutions (P < 0.01). The application of 2% CX followed
by 17% EDTA had the highest microtensile bond strength

In the present study, the microtensile test was used

because of the possibility of performing the analysis in an
area of approximately 1.0 mm2, producing uniformity in
the stress distribution and contributing to obtaining accurate results (17,18).

Published 2010. This article is a US Government work and is in the public domain in the USA.

Endodontic Irrigants on Bond Strength

A. P. Farina et al.

The bond strength to the dentine of the adhesive

system Clearfil SE Bond was affected by the different
endodontic irrigants tested in the present study, confirming the hypothesis under study. Application on the
dentine surface of 1% NaOCl, 1% NaOCl followed by
17% EDTA, 2% CX and 17% EDTA presented statistically
lower bond strength than the control group and the 2%
CX Group, followed by 17% EDTA. However the application of 2% CX followed by 17% EDTA resulted in
higher values of bond strength to the dentine, in addition
to the values being higher than those of the control group
that received no irrigant applications; the limitations of
this study, as is in vitro, may have caused values unexpected for this group. The limitations of this study, as it is
in vitro, may have caused unexpected values in this group.
NaOCl is a halogenated compound, routinely used in
endodontics, which has low surface tension, antiseptic
ability, partially neutralises the toxic products of root
canals and dissolves organic tissue (19). According to our
study, this antimicrobial agent compromised the bond
strength of the adhesive system to dentine. It is thought
that NaOCl leads to oxidation of some of the components
in the dentine matrix, particularly collagen (20,21),
forming protein-derived radicals (22) that compete with
the propagating vinyl free-radicals generated by the
photo-activation of resin adhesives, resulting in premature chain termination and incomplete polymerisation
(23). Besides, NaOCl liberates sodium chloride and
oxygen, and the oxygen causes strong inhibition of the
polymerisation in the adhesive system (24).
The residual irrigant solutions and their products are
likely to diffuse into dentine. As it contains acid monomers in its composition, the self-etching adhesive system
(Clearfil CE Bond) produces increased dentin permeability and increases the flow of fluids into the dentinal
tubules. Thus, the residual chemical substances in the
fluid may interact with the adhesive system and affect the
light polymerisation of the monomer in the demineralised dentine, causing a decrease in bond strength (25).
The chlorhexidine has good antimicrobial activity and
acts on the cell walls of microorganisms causing disarrangement in the intracellular components (11). When
this solution was used separately, it promoted significant
reduction in the bond strength of the adhesive system to
dentine. Edermir et al. (26) reported that endodontic irrigation with CX solution significantly increased bond
strength to root dentine. These authors suggested that
adsorption of CX by dentine may favour resin infiltration
into dentinal tubules. In the present study the increase in
bond strength with the use of CX only occurred after the
use of EDTA. This suggests that the effect of EDTA
increases the bond strength, mainly because of the
surface effect of some chemical reaction. Therefore, EDTA
4

is a powerful anti-oxidant via redox reaction, allowing

free radical polymerisation without premature chains or
failures. Moreover, it has the ability to remove the smear
layer, which could facilitate resinous monomer penetration into the dentinal tubules (27).
The lowest bond strength values were obtained in the
group in which only EDTA was used, and it seems to be
related to the pattern of dentine surface erosion these
solutions are capable of causing (28,29) when used separately. Santos et al. (7) obtained lower bond strength
values for the Clearfil SE Bond self-etching adhesive
system when 5.25% NaOCl was used combined with
17% EDTA than when only NaOCl was applied on
dentine.
Until recently, endodontically treated teeth were
usually treated with a crown, core or dowel (30);
however, when they have a sufficient amount of complete coronal structure they can be restored with an adhesive system and resin composite (13,15). It is known that
a better bond of restorative materials to dentine walls
increases the possibility of marginal sealing, mechanical
resistance to mastication stress and durability of restorations (31).

Conclusion
The bond strength can be affected by endodontic irrigating solutions, as shown by the results of this study.
Further studies should be conducted with regard to the
effect of these irrigating solutions on bond of the adhesive
system to the dentine substrate.

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Published 2010. This article is a US Government work and is in the public domain in the USA.

Endodontic Irrigants on Bond Strength

A. P. Farina et al.

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Published 2010. This article is a US Government work and is in the public domain in the USA.