A comparative study of the removal of smear layer
by three endodontic irrigants and two types of laser
F. H. Takeda, T. Harashima, Y. Kimura & K. Matsumoto
Department of Endodontics, Showa University School of Dentistry, Ohta-ku, Tokyo, Japan
Abstract
Takeda FH, Harashima T, Kimura Y, Matsumoto K.
A comparative study of the removal of smear layer by three
endodontic irrigants and two types of laser. International
Endodontic Journal, 32, 32±39, 1999.
Aim The effects of three endodontic irrigants and two
types of laser on a smear layer created by hand instru-
mentation were evaluated in vitro in the middle and
apical thirds of root canals.
Methodology Sixty human mature extracted
mandibular premolar teeth with a single root canal
and a closed apex were distributed randomly into five
groups of 12 teeth each. Whilst cleaning and shaping
up to a size 60 master apical file with a step-back
technique, the root canals were irrigated with 3mL of
5.25% NaOCL and 3% H2O2, alternately, between each
file size. Group 1 (G1) were control specimens that
were irrigated with a final flush of 17% EDTA. The
teeth in group 2 (G2) were irrigated with a final flush
of 6% phosphoric acid, and group 3 (G3) with 6%
citric acid. In the specimens of group 4 (G4) the root
canals were irradiated with a carbon dioxide (CO2)
laser, and specimens of group 5 (G5) were irradiated
using an Er:YAG laser. The teeth were split longitudin-
ally and prepared for examination by scanning
electron microscopy.
Results Control specimens (G1) showed clean root-
canal walls with open dentinal tubules in the middle
one-third, but in some specimens thick smear layer
was observed in the apical one-third. Specimens
Correspondence: Professor Koukichi Matsumoto, Department of
Endodontics, Showa University School of Dentistry, 2-1-1 Kitasenzoku,
Ohta-ku, Tokyo 145-8515, Japan.
32 International Endodontic Journal, 32, 32±39, 1999
irrigated with a final flush of 6% phosphoric acid
(G2) or 6% citric acid (G3) were cleaner than with
17% EDTA, showing very clean root canal surfaces
in the middle one-third but in the apical one-third
the smear layer was not completely removed,
especially at the openings of the dentinal tubules.
The specimens irradiated with the CO2 laser (G4)
showed clean root-canal walls with the smear layer
absent, charred, melted, recrystallized and glazed in
both middle and apical thirds. The root-canal walls
of the specimens irradiated with the Er:YAG laser
(G5) revealed an absent smear layer with open
dentinal tubules in the middle and apical thirds.
Statistical analysis showed no significant difference in
the cleanliness of root-canal wall between G1 and
G2, and G1 and G3. However, there were statistically
significant differences (P<0.01) between G1 and G4,
and G1 and G5 in the cleanliness of the middle and
apical one-thirds of the root canals.
Conclusions Irrigation with 17% EDTA, 6%
phosphoric acid and 6% citric acid did not remove all
the smear layer from the root-canal system. In
addition, these acidic solutions demineralized the
interbular dentine around tabular openings, which
became enlarged. The CO2 laser was useful in
removing and melting the smear layer on the instru-
mented root-canal walls and the Er:YAG laser was the
most effective in removing the smear layer from the
root-canal wall.
Keywords: citric acid, CO2 laser, Er:YAG laser,
phosphoric acid, smear layer
Introduction
The success of root canal treatment depends on the
root canal system being thoroughly cleansed and
q 1999 Blackwell Science Ltd

disinfected, followed by the adequate obturation of
this space. McComb & Smith (1975) were the initial
investigators to show the presence of a smear layer
in instrumented root canals. Since their study, the
literature is replete with numerous reports utilizing
various methods to remove this smear layer. Smear
layer created during the instrumentation is composed
of tooth structure and some nonspecific inorganic
contaminants. The organic components may consist
of reacted coagulated proteins, necrotic or viable
pulp tissue, odontoblastic processes, plus saliva, blood
cells, and microorganisms (Czonstkowsky et al.
1990). Different irrigant solutions have been used to
remove the smear layer. Sodium hypochlorite
(NaOCl), in a 1±5.25% concentration is an irrigant
solution used widely in root canal treatment because
of its bactericidal properties and ability to dissolve
organic tissues (Yamada et al. 1983, Baumgartner &
Mader 1987); but NaOCl has not been shown to be
effective in removing the smear layer (Prati et al.
1994). Decalcifying solutions such as phosphoric
acid, citric acid, and EDTA have been reported as
suitable for removing the smear layer (Wayman et al.
1979, Francischone et al. 1984, Aktener & Bilkay
1993). On the other hand, studies have shown that
a combination of NaOCL and EDTA removed the
smear layer only partially (Ciucchi et al. 1989, Sen et
al. 1995).
The effects of laser irradiation in endodontics have
been investigated previously. An argon laser had
efficient cleaning activity on the instrumented root
canal surfaces (Harashima et al. 1997a). The
Nd:YAG laser was used to irradiate root canal walls
and caused melted, recrystallized, and glazed surfaces
(Dederich et al. 1984). The Nd:YAG laser was able to
produce clean root canals when combined with hand
filling and showed a general absence of smear layer
and tissue remnants on the root canal wall (Goodis
et al. 1992). The CO2 laser system has been used to
remove organic tissue from the root canal, to fuse
hydroxyapatite, and to open dentinal tubules (Onal et
al. 1993). It was observed in a recent study, that
after Er:YAG laser irradiation most of the debris and
smear layer on the root canal wall were removed,
and dentinal tubules were patent (Takahashi et al.
1996, Takeda et al. 1998).
The purpose of this study was to compare and
contrast the efficacy of 6% phosphoric acid, 6% citric
acid, CO2 laser irradiation, and Er:YAG laser
irradiation on removing the smear layer from the
prepared root canal wall.
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Takeda et al. Study on the smear layer removal
Materials and methods
Sixty recently extracted human mature permanent
mandibular premolar teeth with a single root canal
and a closed apex were used for this study. The teeth
were radiographed to confirm root canal patency and
the absence of a complicated root canal anatomy, and
the crowns were removed at the level of the cementoe-
namel junction. The working length of each root canal
was established 1 mm short of the apical foramen with
a size 15 K-type file. The root canal was cleaned and
shaped up to a size 60 K-type file at the working length
and flared using a step-back preparation technique.
The root canals were irrigated with 3 mL each of
5.25% NaOCl and 3% hydrogen peroxide (H2 O2 )
alternately between each file (Svec & Harrison 1977).
The irrigant solution was delivered with a hypodermic
syringe and a 25-gauge needle as apically as possible
without binding. Finally, irrigation with 3 mL of 5.25%
NaOCl solution, followed by a final rinse with 3 mL of
distilled water, was used to avoid the development of
NaCl crystals. The root canals were then dried with
absorbent paper points. All teeth were distributed
randomly into five groups of 12 teeth each.
Group 1 served as the control. In this group the root
canals were irrigated for 5 min with 10 mL of 17%
EDTA as a final flush, after which the root canals were
irrigated with 3 mL of 5.25% NaOCl followed by a final
rinse with 3 mL of distilled water. In the teeth in group
2, the root canals were irrigated for 5 min with 10 mL
of 6% phosphoric acid as a final flush, after which the
root canals were irrigated with 3 mL of 5.25% NaOCl
followed by final rinse with 3 mL of distilled water. In
group 3, teeth were treated in the same way as group
2 except 6% citric acid was used as a final flush.
Specimens from group 4 were irradiated at a
wavelength of 10.6 mm with a CO2 laser (Opelaser-
03S; Yoshida, Tokyo, Japan) using an output of 1 W,
in continuous mode. The conical tip was introduced
into the root canal, to allow contact with the wall and
irradiation carried out for 3 s at the apex. The laser
was activated during withdrawal strokes from apex to
the orifice of each root canal. Four exposures of 5 s
duration each, a total exposure of 23 s per canal were
made. The teeth from group 5 were irradiated with an
Er:YAG laser through an optical fibre (diameter
0.5 mm), at a wavelength of 2.94 mm (prototype,
Luxar Co, Bothell, WA, USA) with an output of 1 W, a
pulse energy of 100 mJ, and a pulse frequency of
10 Hz. The fibre tip was introduced to the working
length parallel to the root canal wall and placed in
International Endodontic Journal, 32, 32±39, 1999 33
 Study on the smear layer removal Taleda et al.

Figure 1 Group 1 (control), irrigated with a final flush of Figure 4 Smear plugs were observed in the openings of the
17% EDTA. Middle third of root canal wall appeared clean dentinal tubules in the apical one-third of specimens irrigated
with no smear layer, and smear plugs were observed in some with phosphoric acid (original magnification 1000).
specimens (original magnification 1000).

Figure 5 Specimens were irrigated with a final flush of 6%

Figure 2 Control group. In some specimens in the apical
citric acid. The root canal walls in the middle one-third free of
one-third a moderate smear layer remained on the surface of
a smear layer are observed and the dentinal tubules are open
the root canal (original magnification 1000).
(original magnification 1000).

Figure 3 Specimens irrigated with a final flush of 6%

phosphoric acid. In the middle one-third the smear layer was Figure 6 Some specimens irrigated with citric acid show a
removed and tubule openings were enlarged (original moderate smear layer in the apical one-third (original
magnification 1000). magnification 1000).

34 International Endodontic Journal, 32, 32±39, 1999 q 1999 Blackwell Science Ltd

Figure 7 Root canal wall of middle one-third after CO2 laser
irradiation. Open dentinal tubules with smear layer removal
are evident (original magnification 1000).
Figure 8 Specimen in the apical one-third irradiated with
CO2 laser showing a clean root canal wall with smear layer
missing, or charred, melted, recrystallized, and glazed (original
magnification 1000).
contact with the wall and the laser was activated for
3 s at the apex. Four additional laser exposures each of
3 s duration were activated, the fibre tip being
withdrawn from the apex to the orifice along the root
canal wall for a total exposure of 15 s per canal within
the root canal. A water coolant spray was used during
irradiation.
The teeth were split in half after two parallel longitu-
dinal grooves were made with diamond burs on the
outer surface of the root. These did not penetrate the
root canal. The specimens were dehydrated using a
series of graded ethanol solutions (70, 80, 90, 100%),
and coated with platinum after drying.
All specimens were viewed in the middle and apical
thirds of the root canal for evaluation of the cleansing
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Takeda et al. Study on the smear layer removal
Figure 9 Specimen irradiated with the Er:YAG laser. The
smear layer was removed, and then dentinal tubules were
clean and open in middle one-third (original magnification
1000).
Figure 10 In the apical one-third the smear layer was
missing exposing the orifices of the dentinal tubules in
specimens irradiated with the Er:YAG laser (original
magnification 1000).
technique. The 12 representative areas for each group
were photographed with a scanning electron
microscope (JSM-T220A; JEOL, Tokyo, Japan) at
1000 magnification. The photographs were
evaluated for the presence of a smear layer. The rating
system used (Takeda et al. 1998) is shown in Table 1.
The degree of evaluation was scored in a blind manner
based on a four grade scale as shown in Table 1 by a
technician who was not informed of the true nature
and purpose of these experiments. Thus, the judgement
was kept blind.
Statistical analyses of the results were conducted
using the Kruskal Wallis test to determine if there were
significant differences between groups. The Mann-
International Endodontic Journal, 32, 32±39, 1999 35
 Study on the smear layer removal Taleda et al.
Table 1 The rating system
Score Contents
0 No smear layer, open dentinal tubules, smear layer
was completely removed or melted
1 Moderate smear layer, outlines of dentinal tubules
observable, removed or melting in some areas
2 Thin smear layer covering the surface outline of
dentinal tubules which were not discernible, and the
location of the tubule was indicated by a crack,
scattered laser removed or melting
3 Heavy smear layer, outlines of tubules obliterated, no
visible laser removed or melting
Table 2 Remaining smear layer on the root canal walls after
use of different irrigant solutions and laser devices
Group
(N ˆ 12) Middle area Apical area
1 0:917  0:289 1:083  0:515
2 0:500  0:522 1:250  0:754
3 0:417  0:515 1:167  0:577
4 0:167  0:389* 0:333  0:492*
5 0:083  0:289* 0:167  0:389*
Values are shown as mean  SD. *shows a significant difference
(P < 0:01) compared with G1 (control)
Whitney test was used to confirm the significant
differences between pairs of groups.
Results
Group 1 (control)
Specimens irrigated with 5.25% NaOCl, 3% H2 O2 and
17% EDTA as the final flush showed root canal walls
generally cleaned in the middle one-third, with no
smear layer, and only smear plugs in some specimens
(Fig. 1). In the apical one-third apparently open
dentinal tubules were visible which were regularly
distributed. In some specimens a smear layer
remained on the root canal surface (Fig. 2).
Group 2 (6% phosphoric acid)
In specimens treated with phosphoric acid, the smear
layer was removed totally, in the middle one-third
revealing the collagen network of a softened
intertubular dentine around enlarged tubule openings
(Fig. 3). However, in some cases a moderate smear
layer could be observed in the openings of the
dentinal tubules in the apical one-third (Fig. 4).
36 International Endodontic Journal, 32, 32±39, 1999
Group 3 (6% citric acid)
The smear layer was removed from the middle one-
third and the dentinal tubules were open, similar to
the samples treated with 6% phosphoric acid, with
enlarged tubule openings (Fig. 5). In some
specimens a moderate smear layer was seen in the
apical one-third (Fig. 6).
Group 4 (CO2 laser)
In the middle one-third, the root canal wall showed
no smear layer and open dentinal tubules (Fig. 7).
In the apical one-third there were areas with
charred, melted, recrystallized, and glazed smear
layer (Fig. 8).
Group 5 (Er:YAG laser)
Specimens irradiated with the Er:YAG laser showed
no smear layer. Most of the smear layer was
removed, and the dentinal tubules were clean and
open in the middle one-third of the root (Fig. 9). In
the apical one-third the smear layer was lost,
exposing the orifices of the dentinal tubules (Fig.
10).
The incidence of smear layer-free surfaces of the five
groups is given in Table 2. Statistical analysis showed
no significant differences in cleanliness of the smear
layer between G1 (17% EDTA) and G2 (6% phosphoric
acid), and G1 (17% EDTA) and G3 (6% citric acid)
(P > 0:01†: However, there were statistical significant
differences (P < 0:01† between G1 (17% EDTA) and
G4 (CO2 laser), and G1 (17% EDTA) and G5 (Er:YAG
laser) in the cleanliness of the middle and apical one-
thirds of the root canals.
Discussion
In infected root canals, the smear layer produced by in-
strumentation should be removed, because bacteria
may have invaded dentinal tubules and accessory
canals, and the smear plugs produced during instru-
mentation should be removed to facilitate the antibac-
terial effect of intracanal medicaments (Bystrom et al.
1985, Perez et al. 1993). The smear layer constitutes a
negative influence on the sealing ability of obturated
canals, since it is a porous and weakly adherent
interface between the obturation material and the
dentine wall (White et al. 1984, Kennedy et al. 1986).
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The results of this study confirm previous reports
that a final flush with EDTA, which has the potential
of removing the smear layer, did not produce the
expected smear-free surfaces in the apical one-third of
the canal (Ciucchi et al. 1989).
The combination of phosphoric acid and citric acid
removed the smear layer from both the middle and the
apical thirds of the root canal, but it decalcified and
softened the root dentine to a depth of 10 to 15 mm
(Garberoglio & Becce 1994). In addition, the low pH
(1.5) of the solution could have adverse effects on the
periapical tissues. EDTA (17%) removed the smear
layer and, although the dentine was deminerilized, this
did not occur to the same extent as with the acidic
solutions. EDTA changes pH during deminerilization,
and the effect is self-limiting, as the pH decreases, both
the rate of dentine demineralization and the amount of
dentine dissolved decrease (Garberoglio & Becce 1994).
The effects on the removal of the smear layer obtained
with citric acid are similar to those by EDTA (Goldman
et al. 1981), but citric acid is less cytotoxic to tissue
than EDTA (Ando 1985).
The use of the laser has certainly shown great
promise in root canal therapy and the main application
is to remove the smear layer remaining on the instru-
mented root canal walls. The Nd:YAG laser irradiation
produced very clean root canal walls with debris and
smear layer being removed or melted, fused, and recrys-
tallized (Harashima et al. 1997b). The CO2 laser has the
ability to eliminate microorganisms from the root canal
(Zakariasen et al. 1986) and CO2 laser energy has
resulted in increased permeability in coronal dentine by
melting away the smear layer resulting in opening of the
tubule orifices (Pashley et al. 1992). The Er:YAG laser
irradiation has produced melted and sealed dentinal
tubules, accompanied by removal of the organic matrix,
resulting in reduction of fluid permeability, sterilization
of the contaminated root apex, and increased resistance
to root resorption (Paghdiwala 1993).
These two types of lasers (CO2 , Er:YAG) showed the
ability to remove the smear layer, and the surfaces
presented a specific characteristic in each of the laser
types. The root canal walls irradiated by the CO2 laser
showed that all of the root canal surfaces presented a
clean wall with charred, melted, recrystallized, or
glazed smear layer. The Er:YAG laser produced another
characteristic when the root canal wall showed clean
surfaces, free of a smear layer with open dentinal
tubules without any melting. The times used in this
study were 23 sec for CO2 laser and 15 sec for Er:YAG
laser, which were the minimum times to obtain the
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Takeda et al. Study on the smear layer removal
most effective result for each laser (unpublished
observation). If lasers were irradiated for a longer time,
the thermal damage to periapical tissues may occur.
When comparing and contrasting the CO2 , Nd:YAG,
and Er:YAG lasers when irradiated in the teeth, it has
been demonstrated that the Er:YAG laser caused less
thermal damage than either the Nd:YAG or CO2 laser
(Wigdor et al. 1993). Although the temperatures on
the root surfaces were not monitored in our study,
some studies demonstrated that the healthy
surrounding periodontal tissues are not damaged
thermally if the laser equipment is used with a correct
adjustment and temperature increase of the root
canals remains less than 5 8C (Behrens et al. 1993,
Miserandino et al. 1993, White et al. 1994, Ramskold
et al. 1997).
The canals were prepared to a size 60 K-type file,
which was done to allow access for the laser tips. This
was large, but in future laser tips will be made smaller
and enlarging the root canal excessively will be
unnecessary.
The present study demonstrated that phosphoric acid
and citric acid failed to clean the root canal system fully,
and left some minor remnants of smear layer behind.
These acids were used to irrigate for 5 min, which is a
long time for the majority of practitioners, but this time
was the maximum to obtain the most effective results of
these acids with minimum deminerilization judging by
SEM (unpublished observation). Endosonics as a delivery
system for irrigating solutions was not used in this
study, but it has been shown to be a time-saving
procedure and gives cleaner canals in the coronal and
middle thirds when compared with hand preparation.
The major disadvantages are: the cleaning rate is low in
the apical third, transportation sometimes occurs with
straightening of curved root canals and superficial
vertical scratches have also observed at the canal walls
(Haikel & Alleman 1988).
There was no significant statistical difference
between the acid groups and the control group. The
results showed that Er:YAG laser was the most
effective in cleaning prepared root canals but there was
no significant difference between G4 (CO2 laser) and
G5 (Er:YAG laser). There were, however, significant
differences between the control group (unlased) and
lased groups in vitro.
These results suggest that two lasers, especially
Er:YAG laser, are useful for removal of smear layer
from root canals in vivo, but further study on thermal
effects is needed before clinical usage can be
recommended.
International Endodontic Journal, 32, 32±39, 1999 37
 Study on the smear layer removal Taleda et al.
Conclusions
EDTA (17%) was not effective in removing the smear
layer when used as a final flush in instrumented root
canals. The 6% phosphoric acid or 6% citric acid used
as final flush in instrumented root canals did not
remove the smear layer completely. Acidic solutions
markedly deminerialized the dentine. CO2 and Er:YAG
lasers were more efficient in removing the smear layer
than the EDTA and the acidic solutions.
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