ISSN 1054660X, Laser Physics, 2010, Vol. 20, No. 7, pp. 1659–1666.

LASER METHODS IN CHEMISTRY,

© Pleiades Publishing, Ltd., 2010.
Original Text © Astro, Ltd., 2010. BIOLOGY, AND MEDICINE

Increased Viability of OdontoblastLike Cells

Subjected to LowLevel Laser Irradiation1
C. F. Oliveiraa, F. G. Bassob, E. C. Linsc, C. Kurachic, J. Heblinga,
V. S. Bagnatoc, and C. A. de Souza Costaa
a
UNESP—Universidade Estadual Paulista, Araraquara School of Dentistry, Araraquara, SP, 14801903 Brazil
b UNICAMP—Universidade de Campinas, Piracicaba School of Dentistry, Piracicaba, SP, 13414903 Brazil
c USP—Universidade de São Paulo, Physics Institute of São Carlos, São Carlos, SP, 13560970 Brazil

email: casouzac@foar.unesp.br
Received January 28, 2010; in final form, February 18, 2010; published online June 3, 2010

Abstract—Studies have shown that the increase of cell metabolism depends on the low level laser therapy
(LLLT) parameters used to irradiate the cells. However, the optimal laser dose to upregulate pulp cell activity
remains unknown. Consequently, the aim of this study was to evaluate the metabolic response of odontoblast
like cells (MDPC23) exposed to different LLLT doses. Cells at 20000 cells/cm2 were seeded in 24well plates
using plain culture medium (DMEM) and were incubated in a humidified incubator with 5% CO2 at 37°C.
After 24 h, the culture medium was replaced by fresh DMEM supplemented with 5% (stress by nutritional
deficit) or 10% fetal bovine serum (FBS). The cells were exposed to different laser doses from a near infrared
diode laser prototype designed to provide a uniform irradiation of the wells. The experimental groups were:
G1: 1.5 J/cm2 + 5% FBS; G2: 1.5 J/cm2 + 10% FBS; G3: 5 J/cm2 + 5% FBS; G4: 5 J/cm2 + 10% FBS; G5:
19 J/cm2 + 5% FBS; G6: 19 J/cm2 + 10% FBS. LLLT was performed in 3 consecutive irradiation cycles with
a 24hour interval. Nonirradiated cells cultured in DMEM supplemented with either 5 or 10% FBS served
as control groups. The analysis of the metabolic response was performed by the MTT assay 3 h after the last
irradiation. G1 presented an increase in SDH enzyme activity and differed significantly (MannWhitney test,
p < 0.05) from the other groups. Analysis by scanning electron microscopy showed normal cell morphology
in all groups. Under the tested conditions, LLLT stimulated the metabolic activity of MDPC23 cultured in
DMEM supplemented with 5% FBS and exposed to a laser dose of 1.5 J/cm2. These findings are relevant for
further studies on the action of near infrared lasers on cells with odontoblast phenotype.
DOI: 10.1134/S1054660X10130153

1
1. INTRODUCTION
Low level laser (LLL) interacts with different types
of tissues and LLL therapy (LLLT) is known to mod
ulate various biological processes [1–6]. Several stud
ies have demonstrated that LLL penetration capacity
in the tissues depends on laser wavelength. While red
radiation acts on the cell membrane, near infrared
lasers can penetrate deeply into the tissues, stimulating
the cell functions [7]. The alterations in cell mem
brane potential caused by photon energy produced in
the near infrared spectral region induce photophysi
cal and photoelectrical effects [8], which result in
shock between the cells. These events are translated
intracellulary as an increase in ATP synthesis [9],
stimulation of cell division [10], stimulation of cell
metabolism [10–12], acceleration of tissue healing
including the tendons [12, 13] and photo inativati on
of fungi [14].
The effects of near infrared laser radiation on the
cells have been extensively investigated to determine
the optimal laser dose for biostimulation of each cell
type [15, 16]. The main effects of LLLT on different
1 The article is published in the original.
cell cultures have been described as cell proliferation
[17–20], cell differentiation [21], and increase of
DNA synthesis [22] and protein synthesis [18, 23, 24].
However, the lack of methodological standardization
makes it difficult to establish a comparative analysis of
the results in the literature [6, 20].
In dentistry, LLLT has been widely used to treat
dentin sensitivity [25–28]. However, there are no in
vitro or in vivo studies demonstrating the ideal param
eters for irradiation of pulp cells to stimulate the heal
ing of damaged pulp tissue [15, 16]. Regarding dentin
sensitivity, which affects negatively the life quality of a
number of people around the world, the low irradia
tion of the exposed dentin has been clinically used
with high success rate. It may be suggested that the
metabolism of odontoblast cells that underlie the den
tin may be increased by the laser irradiation [29, 30].
Consequently, these stimulated cells may also secret
more dentin matrix proteins than nonirradiated cells.
The deposition of dentin matrix may reduce the dentin
permeability to prevent or even eliminate the dentin
sensitivity. However, the mechanism of odontoblast
cell response to LLLT, in a molecular level, remains
unclear [31]. Therefore, the purpose of this study was

1659
1660 OLIVEIRA et al.
Experimental and control groups formed according to the
FBS concentrations, LLLT doses and irradiation times
FBS Energy Irradiation
5% 10% density, J/cm2 time
Group 1 Group 2 1.5 1 min 20 s
Group 3 Group 4 5 4 min 29 s
Group 5 Group 6 19 16 min 1 s
Control Control 0 –
Note: N = 12 specimens per group; FBS: fetal bovine serum;
LLLT: low level laser therapy
to evaluate the metabolic response of odontoblastlike
cell line (MDPC23) cultures exposed to different
doses of LLLT using a near infrared InGaAsP diode
laser prototype specifically designed to provide a uni
form irradiation of the wells with preestablished laser
parameters.
2. MATERIAL AND METHODS
2.1. MDPC23 Cell Culture
Immortalized cells of the MDPC23 cell line were
defrost and cultured in Dulbecco’s Modified Eagle’s
Medium (DMEM; Sigma Chemical Co., St. Louis,
MO, USA) supplemented with 10% fetal bovine
serum (FBS; Gibco, Grand Island, NY, USA), with
100 IU/mL penicillin, 100 µg/mL streptomycin, and
2 mM/L glutamine (Gibco, Grand Island, NY, USA)
in an humidified incubator with 5% CO2 and 95% air
at 37°C (Isotemp; Fisher Scientific, Pittsburgh, PA,
USA). The cells were subcultured at every 3 days at a
concentration of 20000 cells/cm2 until an adequate
number of cells were obtained for the study.
2.2. LLLT on the MDPC23 Cells
MDPC23 cells at 20000 cells/cm2 concentration
were seeded in 12 wells of sterile acrylic 24well plates
using plain DMEM supplemented with 10% FBS. The
plates were maintained in the humidified incubator
(Isotemp; Fisher Scientific, Pittsburgh, PA, USA)
with 5% CO2 and 95% air at 37°C for 24 h. Thereafter,
the culture medium was aspirated and fresh DMEM
supplemented with either 5% FBS to induce cell stress
by nutritional deficit [16, 24, 32, 33] or 10% FBS was
applied to the cells for 24 hours. After this period and
immediately before laser irradiation, the culture
medium was renewed, maintaining the FBS concen
trations (5 or 10%) according to the experimental and
control groups shown in table.
The LLL device used in this study was a near infra
red indium gallium arsenide phosphide (InGaAsP)
diode laser prototype (LASERTable; 808 ± 3 nm wave
length, 100 mW maximum power output) specifically
designed to provide a uniform irradiation of the wells
in which the cells were seeded.
The radiation originated from the LASERTable
was delivered on the base of each 24well plate at dif
ferent laser doses (1.5, 5, and 19 J/cm2). Although this
diode laser has an output power of 100 mW, the laser
light reached the MDPC23 cells on the bottom of
each well with a final power of 70 mW. The cells were
irradiated every 24 h totalizing 3 applications during 3
consecutive days [17, 22–34]. After the last irradiation
cycle, the 24well plates were maintained in the
humidified incubator (Isotemp; Fisher Scientific,
Pittsburgh, PA, USA) with 5% CO2 and 95% air at
37°C for an additional period of 3 h [11].
The cells in the control groups received the same
treatment as that of the experimental groups. The
24well plates containing the cells in the control
groups were maintained in the LASERTable for the
same time used for irradiation of the corresponding
experimental groups, though without activating the
laser source. One control group was established for
each experimental condition because the different
periods that the cells remained out of the incubator for
laser irradiation should be simulated in the control
groups.
2.3. Analysis of Cell Viability (MIT Assay)
Twelve specimens of each experimental and control
group were used for analysis of cell metabolism. Cell
metabolic activity was evaluated by succinic dehydro
genase (SDH) activity, which is a measure of the mito
chondria] respiration of the cells. For such purpose,
the methyltetrazolium (MTT) assay was used [35].
Each well with the MDPC23 cells received 900 µL
of DMEM associated with 100 µL of MTT solution
(5 mg/mL sterile PBS). The cells were incubated at
37°C for 4 h. Thereafter, the culture medium
(DMEM; Sigma Chemical Co., St. Louis, MO, USA)
with the MTT solution were aspirated and replaced by
700 µL of acidified isopropanol solution (0.04 N HCl)
in each well to dissolve the violet formazan crystals
resulting from the cleavage of the MTT salt ring by the
SDH enzyme present in the mitochondria of viable
cells. Three 100 µL aliquots of each well were trans
ferred to a 96well plate (Costar Corp., Cambridge,
MA, USA). Cell viability was evaluated by spectro
photometry as being proportional to the absorbance
measured at 570 nm wavelength with an ELISA plate
reader (Multiskan, Ascent 354, Labsystems CE, Lês
Ulis, France). The values obtained from the three ali
quots were averaged to provide a single value. Then,
the inhibitory effect of the different groups on cell
mitochondrial activity was calculated and expressed as
medians.
LASER PHYSICS Vol. 20 No. 7 2010
 INCREASED VIABILITY OF ODONTOBLASTLIKE CELLS
2.4. Analysis of Cell Morphology
SDH enzyme activity, OD
by Scanning Electron Microscopy (SEM)
Two specimens were used for analysis of cell mor
phology by SEM. Sterile 12mmdiameter cover
glasses (Fisher Scientific) were placed on the bottom
of the wells of all experimental and control groups
immediately before seeding the cell lines. After expo
sure to the experimental conditions, the culture
medium was removed and the viable cells that
remained adhered to the glass substrate were fixed in
1 mL of buffered 2.5% glutaraldehyde for 24 h and
postfixed with 1% osmium tetroxide for 1 h. The cells
adhered to the glass substrate were then dehydrated in
a series of increasing ethanol concentrations (30, 50,
70, 95, and 100%) and immersed in 1,1,1,3,3,3hex
amethyldisilazane (HMDS; Acros Organics, Spring
field, NJ, USA) for 90 min and stored in a desiccator
for 24 h. The cover glasses were then mounted on
metallic stubs, sputtercoated with gold and the mor
phology of the surfaceadhered L929 and MDPC23
cells was examined with a scanning electron micro
scope (JEOLJMST33A Scanning Microscope,
Tokyo, Japan).
2.5. Statistical Analysis
Initially, the control groups were compared to each
other in order to determine the influence of the period
that the MDPC23 cells remained out of the incubator
on their metabolism. Twoway ANOVA (period out of
the incubator and FBS concentration) and Tukey’s test
were applied to the SDH enzyme activity data. As the
period out of the incubator had a significant influence
on the SDH enzyme activity, all data were transformed
into percentage of the control, considering the mean
of each control as 100%. These data were analyzed sta
tistically by KruskalWallis complemented by Mann
Whitney tests for pairwise comparisons. A significance
level of 5% was set for all analyses.
3. RESULTS
3.1. Analysis of Cell Viability (MTT Assay)
Part 1: Comparison of controls (nonirradiated
cells). Data of the SDH enzyme production (MTT
assay) by the nonirradiated MPDC23 cells accord
ing to the period out of the incubator and FBS con
centrations are presented in Fig. 1. Statistically signif
icant differences (p < 0.05) were found only for the
factors period out of the incubator and FBS concentra
tion. The interaction between these factors was not sig
nificant (p > 0.05). The effect of the FBS concentra
tion was observed only after keeping the cells for 1 min
20 s out of the incubator. For that period, the highest
SDH activity was observed for the 10% concentration,
which differed significantly from the 5% concentra
tion (p < 0.05).
LASER PHYSICS Vol. 20 No. 7 2010
1661
0.35
5% FBS
0.30 10% FBS
0.25
0.20
0.15
0.10
0.05
0
1 min 20 s 3 min 16 min 30 min
Time out of the incubator
Fig. 1. Mean SDH enzyme activity detected by the MTT
assay (optical density) according to the period that the
nonirradiated cells (control groups) remained out of the
incubator (min) and FBS concentration (% FBS). Error
bar = standard deviation, n = 12. Lowercase letters permit
comparison within each FBS concentration and uppercase
letters within each time out of the incubador. Columns
indicated with the same letter are not statistically different
(Tukey, p > 0.05).
When the effect of the period that the cells
remained out of the incubator was evaluated within
each FBS concentration, statistical significance was
observed only for the 5% concentration. Therefore,
the highest SDH activity was observed 30 min after
removal of the cells from the incubator, though this
value did not differ significantly from those obtained
in 3 and 16min periods, which in turn did not differ
from each other (p > 0.05). The lowest SDH activity
was obtained 1 min 20 s after removal of the cells from
the incubator, but it did not differ significantly from
those obtained in the 3 and 16min periods (p > 0.05).
Statistically significant difference (p < 0.05) was found
only between the periods of 1 min 20 s and 30 min.
Part 2: Irradiation doses. Since the period that the
nonirradiated cells remained out of the incubator
exerted a significant effect on the cell metabolism
when 5% FBS was used, all the absolute values of SDH
activity were transformed into percentage of the
respective control, which was considered as 100%.
This was carried out in order to allow the direct com
parison of the irradiated groups by eliminating the
effect of the permanence of the cells out of the incuba
tor. The percentage data according to the irradiation
dose and concentration of FBS are presented in Fig. 2.
Comparing the irradiated and nonirradiated (con
trol) groups, a significantly lower (p < 0.05) activity of
the SDH enzyme was observed only for the 19 J/cm2
irradiation dose when the culture medium was supple
mented with 5% FBS. Cell irradiation did not influ
ence the SDH enzyme activity when the culture
medium was supplemented with 10% FBS since no
statistically significant difference (p > 0.05) was found
between the irradiated groups and their respective
controls.
1662
160
140
Porcentage of SDH activity
120
100
80
60
40
1.5
Fig. 2. Boxwhisker plot (minimum–median–maximum) of the percentage of SDH enzyme activity (MTT assay) for each group.
Comparison of the irradiation doses within each
FBS concentration demonstrated that the highest
activity of SDH enzyme for the 5% FBS supple
mented medium was detected after the dose of
1.5 J/cm2 while the lowest activity was seen after
19 J/cm2, with a statistically significant difference (p <
0.05) between them. An intermediary activity of the
SHD enzyme was recorded for the cells irradiated with
5 J/cm2, which did not differ statistically from the
doses of 1.5 J/cm2 and 19 J/cm2. When the doses were
compared for the 10% supplemented medium, no sig
nificant differences (p > 0.05) were observed.
3.2. SEM Analysis
In the control groups, a large number of MDPC23
cells remained attached to the glass substrate. Those
cells exhibited spindleshaped morphology and several
thin cytoplasmic processes originating from their
membrane (Figs. 3a, 3b, 3c, and 3d), For the experi
mental groups, no significant morphological alter
ations were observed in either of the FBS concentra
tions, and the results were very similar to those
observed in the control groups (Figs. 4a, 4b, 4c, and
4d).
4. DISCUSSION
The direct or indirect mechanisms of action of the
LLLT on cells and the parameters of the laser treat
ment remains unclear [11, 33, 36], It has been specu
lated that the laser light is first absorbed by the tissue
and then the energy received by the cells is transferred
to intracellular components, promoting photoelectri
cal effects that regulate the cell system by means of a
OLIVEIRA et al.
5% irradiado
~
5% naoirradiado
10% irradiado
~
10% naoirradiado
5 19
Irradiation dose, J/cm2
process known as biomodulation [24, 37]. This pro
cess can be triggered by red and infrared laser radiation
and its penetration thresholds into the biological tis
sues depend not only on laser wavelength, but also on
the optical properties, thickness and reflectance of the
irradiated tissue [33, 38]. These data were corrobo
rated by a current in vivo investigation in which the
refractive index behavior of white mice blood under
lowlevel laser irradiation was assessed. The authors
reported that the refractive index behavior in real time
allows determination of the maximum therapeutic
dose on tissue, cellular, and molecular levels. It was
also demonstrated that the refractive index of human
blood can serve as an objective parameter for estimat
ing the level of inflammatory processes both at the
stage of identification and when studying the dynam
ics of a specific disease [36].
When applied at adequate doses, lasers with wave
length in the near infrared spectral region, such as the
InGaAsP diode laser used in the present study, can
reach the mitochondrial membrane of the cells and be
absorbed by the chromophores. These structures are
photosensitive and comprehend the class of porphy
rines, flavins, mitochondrial cytochromes, plasma
membrane and NADH oxidase system, which has fla
voproteins and cytochrome B [39–42]. This spectrum,
more specifically in the 810–830 nm wavelength
range, when absorbed by the coxidase cytochrome of
the mitochondria [34, 39], promotes rapid modifica
tions in the cascade of intracellular chemical and
physical reactions, leading to metabolic alterations
[43]. These modifications result from the electrons
absorbed by the cytochrome during irradiation and
transferred in the oxidative respiration to the CuA
receptors, and further conducted internally from the
LASER PHYSICS Vol. 20 No. 7 2010
 INCREASED VIABILITY OF ODONTOBLASTLIKE CELLS 1663

20 µm 10 µm
(а) (b)

(c) 20 µm (d) 10 µm

Fig. 3. Panel of SEM micrographs representative of cell morphology in each group, a and b (1.5 J/cm2 control group): MDPC
23 cells adhered to the glass substrate exhibited a spindleshaped morphology with several thin cytoplasmic processes originating
from their membrane; c and d (19 J/cm2 control group): Cells with similar morphology covering the entire glass substrate.

hemea/CuA receptors to the hemea3/CuB central
receptor by means of seven intermediary oxireduc
tion reactions, involving Fe and Cu molecules. This
process induces the reduction of molecular oxygen
and the consequent production of reactive oxygen spe
cies (ROS) [37, 44]. Therefore, the presence of intra
cellular ROS seems to alter the REDOX state of the
cell [44]. At concentrations below the cell damage
threshold, the presence of ROS may cause several ben
eficial effects to the cells, such as proliferation, differ
entiation and regulation of cell survival.
The present study evaluated MDPC23 cell metab
olism in response to different laser doses and subjected
or not to stress induced by FBS restriction (5% FBS).
In group 1 (cells subjected to nutritional deficit and
irradiated with 1.5 J/cm2), there was an increase in the
metabolism of the irradiated cells (SDH enzyme
activity; Fig. 1), characterizing a trend of these cells to
respond positively to biostimulation. This fact has also
been demonstrated by Khadra et al. (2005) [19], who
irradiated fibroblasts cultured in DMEM supple
mented 5% FBS on titanium discs. Those authors
reported an increase in cell proliferation and DNA
expression after 72 h when 1.5 and 3 J/cm2 irradiation
doses were used. In a recent study, Oliveira et al. (2008)
[31] observed the occurrence of mitochondrial clus
tering with a granular aspect concentrated in the peri
nuclear region associated with an increase in cell pro
liferation in cells irradiated with 3 J/cm2. Those
authors concluded that such fact suggests an increase
in cell energy, which can lead to an increase in protein
synthesis and genetic material duplication, indicating
a positive response of the cells subjected to laser irradi
ation. In the present study, the groups irradiated with 5
and 19 J/cm2 showed decrease in cell metabolism,
characterizing cell inhibition, which probably
occurred due to overdose of LLLT. This result is in
agreement with the findings of Moore et al. (2005)
[20], who did not observe proliferation of fibroblast
and endothelial cells cultured in DMEM supple
mented with 10% FBS when these cell types were
exposed to a laser dose of 10 J/cm2.
It has been extensively demonstrated that cells
under stress conditions are more sensitive to the laser

LASER PHYSICS Vol. 20 No. 7 2010

1664 OLIVEIRA et al.

(а) 20 µm (b) 10 µm

20 µm 10 µm
(c) (d)

Fig. 4. Panel of scanning electron micrographs representative of cell morphology in each group, a and b (1.5 J/cm2 irradiated
group): A number of odontoblastlike cells remained attached to the glass substrate and exhibited a morphology similar to that of
the control groups; c and d (5 J/cm2 irradiated group): The cytoplasmic cell membrane exhibits thin cellular processes that seem
to be keeping the cells attached to the glass substrate.

biomodulatory effect [6, 16, 19, 45]. These stress con
ditions may vary from alterations in the oxidative
agents to nutritional deficit (decrease of FBS concen
tration in the culture medium), the latter being more
effectively used to evaluate the effects laser irradiation
on cell metabolism [16, 24, 32, 33]. Therefore, several
researchers have evaluated the effects of laser irradia
tion on cells under stress conditions and have observed
that, in most cases, biomodulation has effective
results. Pereira et al. (2002) [46] reported an increase
in the proliferation of NIH 3T3 cells seeded in a cul
ture medium supplemented with only 2.5% FBS. In
the present experiment, when the two FBS concentra
tions were compared at each irradiation dose, it was
observed that the cells responded more positively by an
increase in the metabolic activity when the culture
medium was supplemented with only 5% FBS, espe
cially when the irradiation dose was set at 1.5 J/cm2.
Similar cell response has been described by Khadra
et al. (2005) [19], who attributed the biostimulating
effects obtained in their study to the nutritional deficit.
AlmeidaLopes et al. (2001) [16] reported that fibro
blasts seeded in culture medium without FBS supple
mentation did not show growth. On the other hand,
those authors demonstrated that addition of 5% FBS
to the culture medium resulted in a cell proliferation
rate high enough to induce a stress condition in vitro.
The findings of the present study are consistent with
those previous investigations [16, 19], justifying the
positive metabolic effects obtained for cells cultured in
culture medium containing 5% FBS, especially with
the lowest irradiation dose (1.5 J/cm2).
The cells cultured in DMEM containing 5% FBS
and irradiated with 5 J/cm2 presented a lower meta
bolic activity than that of the respective control group,
though without statistical significance (p > 0.05), and
the most satisfactory FBS concentration was the same
found with the lower laser dose (1.5 J/cm2). The cells
cultured in 5% FBS supplemented medium and irra
diated with 19 J/cm2 had lower cell metabolism com
pared to its respective control, but no significant dif
ference (p < 0.05) was found either. The same response
pattern was identified in the control group. Karu

LASER PHYSICS Vol. 20 No. 7 2010

 INCREASED VIABILITY OF ODONTOBLASTLIKE CELLS
(1990) [47] reported that the stimulation and inhibi
tion thresholds of cell activities are directly related to
the laser dose and power of the laser. This means that
the same photoreceptor molecules, when stimulated
by the application of an adequate laser dose, induces
the transportation of electrons and the REDOX state is
altered to modulate cell metabolism. However, based
on the results of the present study, it may be speculated
that during cell irradiation with high doses, the
byproducts from photobiostimulation might have
caused damage to the MDPC23 cells, which would
explain the inhibition of cell metabolism and even the
inhibition of cell proliferation.
The results of the present study indicate that the
decrease of FBS concentration did not cause damage
to the cells under any of the tested conditions since the
SEM analysis did not show significant morphological
alterations and the irradiated groups preserved the
characteristics of the control groups. Further in vitro
studies are needed to investigate the possibility of
transdentinal stimulation of MDPC23 cells stressed
by laser irradiation of dentin. This kind of in vitro
study tries to simulate a clinical situation of dentin
exposure in patients with dentin hypersensitivity. The
use of LLL irradiation of pulp cells in the treatment of
dentin hypersensitivity is a very promising therapy in
dentistry.
CONCLUSION
It may be concluded that the cells irradiated with a
laser dose of 1.5 J/cm2 showed an increase in cell
metabolism when subjected to experimentally induced
nutritional deficit (5% FBS). Therefore, according to
the research protocol employed in the present investi
gation, exposure of odontoblastlike cell MDPC23
cultures maintained under nutritional deficit condi
tions to the laser light at 1.5 J/cm2 was a trend towards
more favorable for direct biomodulation.
ACKNOWLEDGMENTS
The authors acknowledge the Fundação de Amp
aro à Pesquisa do Estado de São Paulo—FAPESP
(grant nos. 2007/506463 and BP.DR: 2008/547850)
and the Conselho Nacional de Desenvolvimento Cientí
fico e Tecnológico—CNPq (grant nos. 476137/2006 and
301029/20075) for the financial support.
REFERENCES
1. A. Antunes, V. L. R. Salvador, M. A. Scapin, W. de Rossi,
and D. M. Zezell, Laser Phys. Lett. 2, 318 (2005).
2. D. A. M. P. Malta, M. A. M. Kreidler, G. E. Villa,
M. F. de Andrade, C. R. Fontana, and R. F. Z. Liza
relli, Laser Phys. Lett. 4, 153 (2007).
3. H. Jelínkova, T. Dostálová, M. N e∨ mec, P. Koranda,
P. Šim u° nek, M. Miyagi, Y.W. Shi, and Y. Matsuura,
Laser Phys. Lett. 3, 43 (2006).
LASER PHYSICS Vol. 20 No. 7 2010
1665
4. R. de F. Z. Lizarelli, L. T. Moriyama, J. R. P. Jorge, and
V. S. Bagnato, Laser Phys. 16, 849 (2006).
5. H. Jelínkova, J. Šulc, T. Dostálová, P. Koranda, M. N e∨ 
mec, and P. Hofmanova, Laser Phys. Lett. 6, 222
(2009).
6. M. Khadra, S. P. Lyngstadaas, H. R. Haanaes, and
K. Mustafa, J. Biomed. Mater. Res. 73, 55 (2005).
7. S. Parker, Br. Dent. J. 202, 131 (2007).
8. T. Karu, J. Photochem. Photobiol. B 49, 1 (1999).
9. T. Karu, L. Pyatibrat, and G. Kalendo, J. Photochem.
Photobiol. 27, 219 (1995).
10. H. Kikuchi, T. Sawda, and T. Yanagisawa, Arch. Oral
Biol. 41, 871 (1996).
11. T. Karu, L. V. Pyatibrat, G. S. Kalendo, and R. O. Ese
naliev, Lasers Surg. Med. 18, 171 (1996).
12. M. Koutná, R. Janisch, and R. Veselska, Scr. Med.
(BRNO) 76, 163 (2003).
13. J. L. N. Bastos, R. F. Z. Lizarelli, and N. A. Parizotto,
Laser Phys. 19, 1925 (2009).
14. R. A. Prates, E. G. da Silva, A. M. Yamada, Jr., L. C. Su
zuki, C. R. Paula, and M. S. Ribeiro, Laser Phys. 19,
1038 (2009).
15. A. R. Coombe, C. T. G. Ho, and M. A. Darendeliler,
Clin. Orthod. 4, 3 (2001).
16. L. AlmeidaLopes, J. Rigau, R. A. Zangaro, J. Guidugli
Neto, and M. M. M. Jaeger, Lasers Surg. Med. 29, 179
(2001).
17. N. Grossman, N. Schneid, H. Reuveni, S. Halevy, and
R. Lubart, Lasers Surg. Med. 22, 212 (1998).
18. M. Khadra, P. S. Lyngstadaas, R. H. Haanæs, and
K. Mustafa, Biomaterials 26, 3503 (2005).
19. M. Khadra, N. Kasem, S. P. Lyngstadaas, H. R. Haan
ses, and K. Mustafa, Clin. Oral Implants Res. 16, 168
(2005).
20. P. Moore, T. D. Ridgway, R. G. Higbee, E. W. Howard,
and M. D. Lucroy, Lasers Surg. Med. 36, 8 (2005).
21. N. Aihara, M. Yamaguchi, and K. Kasai, Lasers Med.
Sci. 21, 24 (2006).
22. K. Yamada, J. Jpn. Orthop. Assoc. 65, 787 (1991).
23. S. Hamajima, K. Hiratsuka, M. KiyamaKishikawa,
T. Tagama, M. Kawahara, M. Ohta, H. Sasahara, and
Y. Abiko, Lasers Med. Sci. 18, 78 (2003).
24. A. Stein, D. Benayahu, D. Maltz, and U. Oron, Pho
tomed. Laser Surg. 23, 161 (2005).
25. Y. Kimura, P. WilderSmith, K. Yonaga, and K. Matsu
moto, J. Clin. Periodontol. 27, 715 (2000).
26. F. Schwarz, N. Arweiler, T. Georg, and E. Reich,
J. Clin. Periodontol. 29, 211 (2002).
27. T. M. Ciaramicoli, C. R. R. Carvalho, and P. C. Edu
ardo, Lasers Surg. Med. 33, 358 (2003).
28. E. M. A. Clavijo, V. R. G. Clavijo, M. C. Bandeca,
M. R. Nadalin, M. F. Andrade, J. R. C. Saad, and
A. A. Pelegrine, Laser Phys. 19, 2041 (2009).
29. R. H. Dababneh, A. T. Khouri, and M. Addy, Br. Dent.
J. 187, 606 (1999).
30. G. E. P. Villa, A. B. C. E. D. Catirse, R. C. C. Lia, and
R. F. Z. Lizarelli, Laser Phys. Lett. 4, 690 (2007).
31. C. F. Oliveira, J. Hebling, P. P. C. Souza, N. T. Sacono,
F. R. Lessa, R. F. Z. Lizarelli, and C. A. S. Costa, Laser
Phys Lett. 5, 680 (2008).
1666 OLIVEIRA et al.
32. E. M. V. B. Cagnie, M. J. Cornelissen, H. A. Declerecq,
and D. C. Cambier, Lasers Surg. Med. 18, 95 (2003).
33. Y. L. Jia and Z. Y. Guo, Lasers Surg. Med. 34, 323
(2004).
34. M. Kreisler, A. B. Christoffer, B. Willerstausen, and
B. d’Hoedt, J. Clin. Periodontol. 30, 353 (2003).
35. T. Mosmann, J. Immunol. Methods 65, 55 (1983).
36. S. A. Gonchukov and Yu. B. Lazarev, Laser Phys. 13,
749 (2003).
37. H. Tuby, L. Maltz, and U. Oron, Lasers Surg. Med. 39,
373 (2007).
38. N. MochizukiOda, Y. Kataoka, Y. Cui, H. Yamada,
M. Heya, and K. Awazu, Neurosci. Lett. 323, 207
(2002).
39. H. Kale, P. Harikuman, S. B. Kulkarni, P. M. Nair, and
M. S. Netrawali, Mutat. Res. 298, 17 (1992).
40. G. Y. Fraikin, M. G. Strakhovskaya, and A. B. Rubin,
J. Photochem. Photobiol. B 34, 129 (1996).
41. A. M. Edwards and E. Silva, J. Photochem. Photobiol.
B 63, 126 (2001).
42. T. Karu, Lasers Life Sci. 2, 52 (1988).
43. T. I. Karu, L. V. Pyatibrat, and G. S. Kalendo, Photo
chem. Photobiol. Sci. 3, 211 (2004).
44. R. Lubart, M. Eichler, R. Lavi, H. Friedman, and
A. Shainberg, Photomed. Laser Surg. 23, 3 (2005).
45. M. M. Marques, A. N. Pereira, N. A. Fujihara,
F. N. Nogueira, and C. P. Eduardo, Lasers Surg. Med.
34, 260 (2004).
46. S. N. Pereira, C. P. Eduardo, E. Matson, and
M. M. Marques, Lasers Surg. Med. 31, 263 (2002).
47. T. Karu, J. Photochem. Photobiol. 52, 1089 (1990).
LASER PHYSICS Vol. 20 No. 7 2010