Lasers Med Sci

DOI 10.1007/s10103-015-1757-y

ORIGINAL ARTICLE

Investigations of radicular dentin permeability

and ultrastructural changes after irradiation with Er,Cr:YSGG
laser and dual wavelength (2780 and 940 nm) laser
Tamara Sardar Al-Karadaghi 1,2 & Rene Franzen 1 & Hussein A. Jawad 2 &
Norbert Gutknecht 1

Received: 2 August 2014 / Accepted: 23 April 2015

# Springer-Verlag London 2015

Abstract The aim of this study was to assess the effective-
ness of dual wavelength (2780 nm Er,Cr:YSGG, 940 nm di-
ode) laser in elimination of smear layer comparing it with Er,
Cr:YSGG laser in terms of radicular dentin permeability and
ultrastructural changes of root canal walls. Fifty-one sound
single-rooted extracted teeth were instrumented up to size F4
and divided into three groups: group Co, non-irradiated sam-
ples; group A, irradiated with Er,Cr:YSGG laser; group B,
irradiated with the dual wavelength laser. Afterward, the roots
were made externally impermeable, filled with 2 % methylene
blue dye, divided horizontally into three segments reflecting
the cervical, middle, and apical thirds then examined under
microscope. Using analytical software, the root section area
and dye penetration area were measured, and then, the per-
centage of net dye penetration area was calculated. Addition-
ally, scanning electron microscope investigations were accom-
plished. Analysis of variance (ANOVA) showed significant
differences between all groups over the three root thirds.
Dye permeation in dual wavelength laser group was signifi-
cantly higher over the whole root length: cervical, middle, and
* Tamara Sardar Al-Karadaghi
t.sardar.m@gmail.com
Rene Franzen
rfranzen@ukaachen.de
Hussein A. Jawad
hussein@ilps.uobaghdad.edu.iq
Norbert Gutknecht
ngutknecht@ukaachen.de
1
Department of Conservative Dentistry, Periodontology and
Preventive Dentistry, RWTH University Hospital Aachen,
Pauwelsstrasse 30, 52074 Aachen, Germany
2
Department of Biomedical Applications, Institute of Laser for
Postgraduate Studies, University of Baghdad, Baghdad, Iraq
apical compared to Er,Cr:YSGG laser group and non-
irradiated samples (p<0.001). Scanning electron micrographs
of dual wavelength irradiated samples showed a distinctive
removal of smear layer with preservation of the annular struc-
ture of dentinal tubules. Er,Cr:YSGG laser root canal irradia-
tion produced uneven removal of smear layer, in efficient
cleanliness especially in the apical third. There was no sign
of melting and carbonization. Within the studied parameters,
root canal irradiation with dual wavelength laser increased
dentin permeability.
Keywords Dual wavelength laser . Radicular dentin .
Permeability . Ultrastructure
Introduction
The effective removal of organic and inorganic tissue rem-
nants along the complex root canal system is a principal con-
dition in the success of root canal treatment. These tissue
remnants reduce dentinal permeability, stand as an obstacle
that prohibit the penetration of intracanal medicaments into
dentin, and protect the microorganisms present in deep dentin.
Besides, this smear layer itself may be infected. Chemical
agents like NaOCl and EDTA that are widely conjugated with
mechanical instrumentation have been suggested to indorse
root canal disinfection [1]. A direct contact between chemical
agents and microorganisms is required in order to execute its
bactericidal action. However, the penetration depth of chemi-
cal agent is about one order lower than the penetration ability
of microorganisms that limit their bactericidal action [2]. On
the other hand, studies revealed incomplete removal of the
smear layer from the root canal walls, particularly in the apical
third, when chemo-mechanical preparation was used [3–5].

Due to the distinguished characteristics of lasers: control-
lable deposited energy, ability of penetration into different
thicknesses depending on their wavelengths, lasers have been
widely investigated in root canal treatment.
According to the composition of chemical agents, each act
on radicular dentin in a specific way and presenting a certain
morphological changes on dentin surface. The same standard
can be applied in laser-assisted root canal treatment. Each
laser has a precise property, because they emit different wave-
lengths, promoting a different tissue interaction [6].
Weichman and Johnson were the first users of CO2 laser in
endodontics to seal the apical foramen [7]. Due to its antici-
pated disinfection properties, Nd:YAG laser was widely ac-
cepted in endodontics; however, SEM investigations revealed
an ultrastructural changes: melting of dentin and closure of
exposed dentinal tubules [8–10].
Diode lasers of 810, 940, and 980 nm wavelengths offered
a better penetration to formally inaccessible areas of the tubu-
lar network [11–13], allowing the decontamination action to
deeper layers of dentin. However, the use of these lasers is
usually combined with chemical irrigants due to their limited
ability to remove smear layer.
On the other hand, erbium lasers such as Er:YAG and Er,
Cr:YSGG lasers with wavelengths of 2940 and 2780 nm, re-
spectively, are suitable to remove dental hard tissues due to
their high affinity to water molecules [14, 15]. As a conse-
quence of their high absorption coefficient, germ reduction
would predominantly take place in the main canal, limiting
their disinfection exploit in the lateral tubular system of dentin
[16].
Recently, Faria et al. [17] carried an in vitro study to eval-
uate the effect of 980 nm diode laser on the ultrastructure and
fracture resistance of radicular dentin. They hypothesized that
after 980 nm diode laser irradiation of root canal, a modified
smear layer was observed in specimens that were irrigated
with water and then laser irradiated with 1.5 and 3 W/100 Hz.
In the current research work, the efficiency of combining
the ablation ability of 2780 nm Er,Cr:YSGG laser with good
penetration depth of 940 nm diode laser using radial firing tip
will be thoroughly investigated, regarding dentin permeability
and ultrastructural changes of radicular dentin, using dual
wavelength laser as an adjunct in root canal treatment.
Materials and methods
Sample selection and preparation
Fifty-one sound single-rooted human extracted teeth obtained
from the faculty of Dentistry (Ministry of Health, Iraq) were
stored in distilled water containing 0.1 % thymol had been
used in this experiment. The teeth were scaled with curate,
and the crowns of the whole teeth were sectioned with
Lasers Med Sci
double-face diamond discs (915 DC, Meisinger, Germany)
mounted on slow speed handpiece, standardized to length of
14 mm. Canal orifices were flared with #1 and #2 Gates-
Glidden drills (MailleferDentsply, RJ, Switzerland). The bio-
mechanical preparation of the root canals were accomplished
using nickel–titanium Pro-Taper files to an apical size corre-
spond to F4 Pro-Taper instrument (Dentsply, Switzerland)
1 mm shorter than working length. During the biomechanical
preparation, the teeth were irrigated with 2 ml of distilled
water between files. Final irrigation with 2 ml of distilled
water was performed after final instrumentation to remove
any debris remnant.
Laser irradiation
The dual wavelength laser (Biolase, San Clemente, CA, USA)
is a laser that emits both 2780 nm Er,Cr:YSGG laser and
940 nm diode laser simultaneously. The delivery was via ra-
dial firing laser tip (RFT) fiber (diameter 415 μm, length
21.17 mm, calibration factor 0.85). Panel setting was (Pave =
1.25 W, repetition rate=50 Hz, pulse duration=50 μs) for Er,
Cr:YSG and for diode laser (Pave =2 W), with 60 % water
level and 30 % air pressure. The output power for both lasers
was measured by Field MaxII Top powermeter (Coherent,
USA) using MZ6 laser tip with calibration factor of 1; then,
the measured value was multiplied by the calibration factor of
RFT. The output power from RFT laser tip was calculated to
be 1.06 W for Er,Cr:YSGG laser and 0.51 W for diode laser.
For permeability experiment, the samples were divided
randomly into three groups (n=15):
Group Co: control group of non-irradiated samples.
Group A: specimens were irradiated with Er,Cr:YSGG
laser (Pave =1.06 W, P.R.R.=50 Hz, pulse duration=
50 μs, 60 % water, 30 % air pressure) for one round
comprised three cycles, 18 s of laser irradiation for each
cycle separated by 5 s rest time (total irradiation time of
54 s).
Group B: specimens were irradiated with Er,Cr:YSGG
laser with the same power setting of group A for three
rounds (nine cycles), in combination with 940 nm diode
laser Pave =0.51 W, pulse width=4 ms, 10 ms, equivalent
to a duty cycle of 20 % and 50 % for the second and third
rounds, respectively, total irradiation time of 162 s. An
interval time of 20 s was set between laser rounds.
During laser irradiation, the RFT fiber tip was positioned
1 mm shorter than working length. Irradiation of dentin walls
was performed from apical to cervical direction in a helicoidal
motion at a rate of 1 mm s−1. According to the manufacturer
instructions and methodology of Ramalho et al. and Esteves-
Oliveira et al. [18, 19], the irradiation cycles were performed
three times (one round) for group A with total irradiation time
Lasers Med Sci
of 54 s. The total irradiation time was 162 s for group B with
nine irradiation cycles (three rounds). Before irradiation, the
canal was irrigated with 2 ml of distilled water at room
temperature.
Permeability experiment
After completion of laser irradiation for the two laser groups,
the apex was sealed with sticky wax. The prepared roots were
made impermeable externally with three coats of ethyl cyano-
acrylate, dried, and transferred them individually to a plastic
PCR tube (Eppendorf Vertrieb Deutschland GmbH,
Wesseling-Berzdorf, Germany). The specimens were filled
with 2 % methylene blue dye [19, 20] with the aid of hypo-
dermal syringe then placed inside an incubator (Heraeus Elec-
tronic, Heinrich Faust GmbH, Cologne, Germany) at 37 °C
for 20 min. When this time had interceded, they were rinsed
thoroughly under running water. The root canals were dried
with absorbent paper cones. The samples were sectioned hor-
izontally into three segments reflecting the cervical, middle,
and apical thirds with low-speed diamond saw [21] (Exakt
Apparatebau, Norderstedt, Germany); then, the root section
was washed with distilled water to measure the dye penetra-
tion along the root canal sections. The first 2 mm from the
cervix was cut and excluded from microscopic examination.
Evaluation method
The prepared root sections were observed under light micro-
scope (MZ6 Leica with integrated Hitachi HV-C20A Camera,
Germany) that connected to PC and analyzed via DISKUS
analytical program (Ver. 4.80.9629, Tech. Office Hilgers, Ger-
many). The total area and dye penetration area were measured
by the software for all root thirds (cervical, middle, and apical)
then subtracted from its corresponding root canal area to ob-
tain the net value. The percentage of net dye penetration area
for each third was calculated from dividing the net dye pene-
tration area over the net total area and multiplied by 100 %.
SEM investigation
Ultra morphological investigation was performed using six
canines, prepared in the same way and irradiated. Afterward,
the prepared roots were sectioned longitudinally into two
parts. One hemisection of each root was randomly chosen;
they were dehydrated with a graded series of ethanol: 30 %,
50 %, 70 %, 90 %, and finally with absolute ethanol 100 %.
Afterward, all the samples were dried by critical point method
then gold sputtered and mounted on metal stubs. Initially, the
specimens were analyzed by SEM (XL 30ESEM FEG Com-
pany Fei, Netherlands) first viewed in a panoramic vision, and
later, the scanning electron micrographs were accomplished at
500× and 1199× from a standardized area (cervical, middle,
and apical) of radicular surface.
Results
Permeability experiment
The analysis of variance (ANOVA) demonstrated that there
were significant differences, at a significance level of α=5 %,
among all groups in the cervical, middle, and apical root thirds
(p<0.05).
The post hoc Tukey test showed that in the cervical third,
middle third, and apical third, group B had the highest dye
penetration means with highly statistical significant differ-
ences from group A (Er,Cr:YSGG laser) and control group
(p<0.001) (Fig. 1).
In the same way, there was no significant difference in
means of dye penetration between group A and control group
over the root thirds: cervical, middle, and apical. The mean
percentage of dye permeation was lower in the apical third
section for all experimental groups compared to cervical and
middle thirds. Means of percentage and standard deviations
are shown in Table 1.
Scanning electron microscope
Two micrographs were taken for each third (apical, middle,
and cervical) of specimens of all groups. In the control group,
the root canal walls were generally covered with heavy smear
layer and debris. Especially in the apical root third, cloaking
and obliteration of dentinal tubules by debris as a result of
mechanical instrumentation of root canal were obvious
(Fig. 2).
In group A, smear layer and debris had been removed to-
tally in some area; nevertheless, also some areas showed den-
tinal tubules occlusion by smear layer, inefficient cleanliness
mainly in the apical third of the root. There was no sign of
melting or cracking over the three root thirds (Fig. 3).
The micrographs of group B revealed encrusted irregular
dentinal surfaces in which the smear layer was removed, with
evidently open dentinal tubules, and no signs of melting or
carbonization. Protrusion of dentinal tubules toward the laser
irradiated surface was observed, indicating that intertubular
dentin suffered more ablation than peritubular dentin (Fig. 4).
Discussion
Permeability is a distinctive characteristic of dentin resulted
from the presence of dentinal tubules and depend on the num-
ber and diameter of tubules that spread along the whole width
of dentin in both crown and root portion of the tooth. Changes
 Lasers Med Sci

Fig. 1 Stereomicroscope image

obtained after transversal cuts and
dye solution penetration for
experimental groups corresponds
to root thirds. Cervical third (A),
middle third (B), and apical third
(C). Bar size 20 μm. Original
magnification 25×

may happen to the dentin permeability according to the treat- 0.4×104 cm−1) [25]. This fact would explain the confinement
ment protocol used during endodontic therapy [22]. It has of Er,Cr:YSGG laser in the main channel. On the contrary,
been postulated that after mechanical instrumentation, about 940 nm diode laser is predominantly scattered through the
35 % of root canal surface remains untouched [23, 24]. These inhomogeneous dentin structure rather than absorption which
uncleansed areas, besides being as reservoir for bacteria, may is neither happened in water nor in hydroxyapatite [26, 12].
act as a barrier that impairs the proper root canal filling. Consequently, a high degree of decontamination can be ex-
The results of this study revealed that among the three groups pected, even in deep layers of dentin [11].
studied, group B showed the highest increase in dentin perme- However, many researches stated an increase in the perme-
ability. This finding was confirmed by the scanning electron ability of dentin after exposure to diode laser. These findings
microscope analysis that presented total absence of dentinal were mainly resulted from combining a chemical irrigant like
smear layer. Dentinal tubules were widely open in all root thirds: EDTA or NaOCl with diode laser, taking into account when
cervical, middle, and apical, after irradiation with dual wave- water was set as an irrigant solution with diode laser (similar
length combination. This finding can improve the mechanical to our study) dye penetration that was much lower [27–29].
interlocking of endodontic sealer into root canal dentin surface. Our findings match those of Silva et al. in which Er,
In addition, laser energy can propagate via the open dentinal Cr:YSGG laser with 0.75 W average power produced uneven
tubules enhancing bactericidal action into deep layers of dentin. removal of smear layer. Some area of insufficient cleanliness
As dentin contain about 20 % water by volume, the main and absence of melting and carbonization was observed due to
interaction is absorption of Er,Cr:YSGG laser by water mole- irregular irradiation. Dentin permeability for all root thirds
cules due to its high absorption coefficient (approximately was statistically similar to control group (non-irradiated sam-
ples); meanwhile, increasing the average power to 2.5 W, den-
tin permeability improved to 45.73 % in the apical third [21].
Table 1 Means of percentage of dye penetration and standard Nevertheless, opposite results were observed by Onay et al.,
deviations (SD) in each group over the three root thirds
they verified that the Er,Cr:YSGG laser treatment did not en-
Root thirds Control Group Group A Group B hance the sealing ability of the sealers compared with EDTA
[30]. Also, when Er:YAG laser at 1.5 W average power and
Mean SD Mean SD Mean SD 15 Hz was used to irradiate radicular dentin, Er:YAG laser did
Cervical 17.14 % 9.75 27.68 % 5.69 80.56 %a 18.73 not increase the permeability in the apical third of the root
Middle 18.82 % 10.84 23.09 % 8.24 82.95 %a 16.41
[19]. These findings come in accordance with our results;
Apical 12.52 % 4.98 15.77 % 10.51 67.09 %a 19.80
there was no significant difference between non-irradiated
control group Co and group A, by which the inhomogeneous
a
Statistically significant difference between the groups in each root third irradiation of root canal surface occasioned in insufficient
Lasers Med Sci

Fig. 2 Scanning electron

microscope of control group
radicular dentin, A cervical third,
B middle third, and C apical third
(500×) and a cervical b middle,
and c apical on higher
magnification (1199×)

removal of smear layer. Also, SEM findings for group A did morphology. Er,Cr:YSGG laser created clean dentin surface,
not differ from previous studies regarding surface clearly opened dentinal tubules. As a result of its high affinity

Fig. 3 Scanning electron

microscope of group A radicular
dentin, A cervical third, B middle
third, and C apical third (500×)
and a cervical, b middle, and c
apical on higher magnification
(1199×). Uneven removal of
smear layer can be seen, presence
of some debris (white rhomboid),
obliteration of dentinal tubules
(white arrow), an area of clean
dentin surface and open tubules
(white circle), and presence of
collagen fibers (white square)

Fig. 4 Scanning electron
microscopy of group B radicular
dentin, A cervical third, B middle
third, and C apical third (500×)
and a cervical, b middle, and c
apical on higher magnification
(1199×). Almost complete
removal of smear layer, annular
and open dentinal tubules along
the three thirds of the root
to (–OH) in water molecules, this wavelength ablates
intertubular dentin more than peritubular dentin due to lower
mineral content with absence of cracking, melting, or
resoldification [31, 32].
RFT delivers laser energy in a unique pattern and ensures
more homogenous removal of smear layer, allowing better
removal of smear layer compared to bare fibers [33]. This fact
could explain the distinctive pattern of the dual wavelength in
group B as the irradiation time increased three folds of group
A (Er,Cr:YSGG laser).
After all, during laser-assisted root canal treatment, the
search for a specific wavelength and irradiation protocol are
vital points in the safety and efficiency of the procedure. Fur-
ther investigations regarding the antibacterial efficacy at dif-
ferent depths of dentin are now inspected for this wavelength
combination before bringing this laser to in vivo trials.
Conclusions
Based on the results of this in vitro study, we can conclude that
radicular dentin irradiation with dual wavelength laser (three
rounds, Pave =1.06 W for Er,Cr:YSGG laser, Pave =0.51 W for
diode laser, and 162 s irradiation time) is effective in increas-
ing dentin permeability with superior statistical results when
compared with Er,Cr:YSGG laser group. A smear layer and
debris were removed effectively from root canal walls and
Lasers Med Sci
appear to have an interesting application in laser-assisted root
canal treatment.
Conflict of interest No conflicting financial interests exist.
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