archives of oral biology 52 (2007) 899–903

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Short communication

Effects of low-power red laser on dentine–pulp interface

after cavity preparation. An ultrastructural study

Bruno Miranda Godoy a, Victor Elias Arana-Chavez c, Silvia Cristina Núñez b,

Martha Simões Ribeiro b,*
a
Professional Master Lasers in Dentistry, IPEN-CNEN/SP, São Paulo, Brazil
b
Center for Lasers and Applications, IPEN-CNEN/SP, São Paulo, Brazil
c
Department of Cell and Developmental Biology, ICB-USP, São Paulo, Brazil

article info abstract

Article history: Objective: Studies on the influence of low-power red laser on the repair of dental structures
Accepted 5 February 2007 are very scarce. This study investigated the effects of the laser therapy on the ultrastructure
of the dentine–pulp interface after conservative class I cavity preparation.
Keywords: Design: Two female volunteers with 8 premolars indicated for extraction for orthodontic
Dental cavity preparation reasons were recruited. Class I cavities were prepared and the teeth were randomly divided
Dental pulp into two groups. The first group received treatment with a GaA1As laser, l = 660 nm, power
Laser therapy of 30 mW and energy dose of 2 J/cm2, directly and perpendicularly into the cavity in a single
Odontoblasts visit. After the irradiation, the cavities were filled with composite resin. The second group
Transmission electron microscopy received the same treatment, except by the laser therapy.
Results: Twenty-eight days post-preparation, the teeth were extracted and processed for
transmission electron microscopy analysis. Two sound teeth, without cavity preparation,
were also studied. The irradiated group presented odontoblast process in higher contact
with the extracellular matrix and the collagen fibrils appeared more aggregated and
organised than those of control group. These results were also observed in the healthy teeth.
Conclusion: These findings suggest that laser irradiation accelerates the recovery of the
dental structures involved in the cavity preparation at the predentine region.
# 2007 Elsevier Ltd. All rights reserved.

1. Introduction factors including heat provoked during cavity preparation,

method of placement of the restorative material, chemical
Dental restorative procedures are a source of injury to the pulp irritants, among others.1
tissue. In many circumstances, a caries lesion does not Dentinogenesis is a continuous and dynamic process
represent the biggest threat to the pulp tissue; but instead, regulated by a single layer of highly differentiated post-mitotic
the surgical techniques and materials employed to restore the odontoblast cells.1 Dentine matrix secreted by odontoblasts
tooth configuration may represent the source of aggression. is the major constituent of the dental mineralized tissue
The aggression to the pulp tissue may occur due to several being composed of inorganic and organic components. The

* Corresponding author. Tel.: +55 11 3816 9313; fax: +55 11 3816 9315.
E-mail address: marthasr@usp.br (M.S. Ribeiro).
0003–9969/$ – see front matter # 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.archoralbio.2007.02.003
900 archives of oral biology 52 (2007) 899–903

inorganic components are mostly hydroxyapatite and water. the teeth. Standard cavity preparations were cut into occlusal
Collagen fibrils and non-collagenous proteins are the organic dentine with a diamond bur using the least pressure at a drill
segment secreted by odontoblasts.2,3 speed of 4000 rpm with water spray coolant. Regular 2 mm
The objective of a regenerative pulp treatment is to width  5 mm depth cavities were performed, finishing at
reconstitute normal tissue continuum at the pulp-dentine dentine level, without pulp exposure. Since all elements were
border, regulating tissue-specific processes of tertiary denti- free of dental caries prior to preparation, the depth of the
nogenesis. Tertiary dentine is synthesized as a response to cavities could be standardised.
various external stimuli, such as dental caries, attrition and The teeth were washed and cleaned with pumice and
trauma and may be divided into two sub-categories, the water. The samples were, then, divided into two groups. The
reactionary dentine and the reparative dentine.2 The reac- laser group (LG) had the cavity irradiated with the continuous
tionary dentine as well as the reparative dentine is the best emission GaAlAs (l = 660 nm) diode laser (LLLT-Biowave-
protection material to dental pulp since the tubular pattern of Kondortech, S. Carlos, Brazil), 2 mm beam diameter with a
the reparative dentine varies from a discontinuous to an 208 of beam divergence, maximum output power of 30 mW. A
atubular nature, and thus, the reparative dentin matrix radiant exposure of 2 J/cm2 was applied.5,11 The beam was
permeability is reduced and diffusion of noxious agents from applied directly into the pulpal wall of the cavity for 2 min and
the tubules is prevented.4 15 s. The teeth were restored with composite resin (Z250, 3 M,
Therapeutic effects obtained with low-power laser are used St. Paul, MN, USA) according to manufacturer’s instructions.
for several endings in dentistry, such as acceleration of the The control group (CG) had cavities prepared and restored as
healing process, bone regeneration, attenuation of aching previously described. The teeth in this group did not receive
processes, tooth decay prevention, etc.5–10 Some works laser irradiation.
suggest the biomodulation influence of laser radiation on Two healthy teeth remained without cavity preparation
the inflammatory and reparative process in pulp tissue, and were also analysed.
showing that low-intensity laser therapy (LILT) can be used Teeth of each group were extracted under local anaes-
as a helper on tertiary dentine formation.11 In the aforemen- thesia 28 days after the procedure. The teeth were cleaned
tioned works, the low-intensity laser radiation was used after and all soft tissues covering the root surface were carefully
cavity preparations with pulp exposition. Such preparation is removed. The samples were immediately immersed in 2%
aggressive and it is not the major cavity preparation glutaraldehyde plus 2.5% formaldehyde buffered with 0.1 M
performed in dental offices on a daily basis. sodium cacodylate, pH 7.4. Specimens were immersed in a
It is unclear which cavity preparation depth would promote beaker containing 40 mL of fixative at room temperature,
little or no pulp injury. Moreover, it is also unclear the pulp which was placed in a 20 cm  20 cm glass recipient filled
response to different cavities preparation depth and to with ice and placed in a Pelco 3440 laboratory microwaves
different restorative materials.12 Besides the depth of the (MW) oven (Ted Pella; Redding, CA). The temperature probe of
cavity preparation, the dentinal tubule permeability, as the oven was submersed in the fixative and the specimens
aforementioned, is also an important factor, since tubule were exposed to MW irradiation at a 100% setting for three
permeability allows the progression of caries, bacterial periods of 5 min with the temperature programmed to a
leakage and chemical irritants towards pulp tissue.13 The maximum of 37 8C.14 After MW irradiation, the samples were
best protection of the pulp tissue is offered by the dentine transferred to fresh fixative and remained in it overnight at
secreted by the odontoblasts. So far, there is not an artificial 4 8C. The specimens were washed in 0.1 M sodium cacodylate
material that can be placed into a tooth that would provide buffer, pH 7.2 for 1 h. Decalcification was made in an aqueous
better protection to the pulp. Therefore, the preservation of solution of 4.13% EDTA, replaced every 48 h, during approxi-
the odontoblast cells must be a goal of the restorative dental mately 6 months.
treatment. The samples were dehydrated in graded concentrations of
Thus, the proposal of this paper is to evaluate ultrastruc- ethanol and embedded in Spurr resin. Toluidine blue-stained
turally, by transmission electron microscopy, the effects of red 1-mm-thick sections were analysed in a light microscope and
emission laser on the pulp–dentine interface after conserva- regions of the interface pulp–dentine were selected for
tive cavity preparation and restoration with composite resin ultrathin sections. Ultrathin cuts (80 nm) were made from
without pulp exposition, looking for any beneficial effects of the chosen region by using a diamond ultramicrotome (Leica
this therapy on the dentin repair process. Ultra cut E). The specimens were analysed in a transmission
electron microscope (JEOL 1010, JEOL Inc., Japan), operating at
80 kV.
2. Material and methods

Ethical approval was obtained from the School of Dentistry- 3. Results

University of São Paulo’s Research Ethical Committee. After
written consent was given, two female patients, aged 24 and The ultrastructural analysis of the interface dentine–pulp, 28
27 years, had six class I cavities prepared in non-carious intact days post-cavity preparation, showed that the teeth irradiated
premolars teeth, which were scheduled for extraction for with red laser presented more organised collagen fibrils and
orthodontic reasons. The patients were anaesthetized, and less retraction in the odontoblast process compared to
pre-operative radiographs were taken. The cavities were prepared non-irradiated premolars. The junctional complexes
prepared under absolute isolation to avoid contamination of among odontoblasts were not altered.
 archives of oral biology 52 (2007) 899–903 901

Fig. 2 – Electron micrograph of predentine region in the

pulp–dentine interface of a premolar without cavity
preparation. It can be observed two odontoblasts (O) and
the junctional region (J) between them. T40,000.

Fig. 1 – Electron micrograph of predentine region in the

pulp–dentine interface of a premolar without cavity restored without irradiation. Morphologically, the lased
preparation. It can be noticed the odontoblast process (O) samples are similar to the healthy specimens without cavity
totally in contact with the collagen fibrils (C) of the preparation. Regarding to junction area, no significant
extracellular matrix. T12,000. differences were observed among the groups.
LILT is being recommended for some authors due to its
analgesic and anti-inflammatory effects over the pulp tissue,
The healthy teeth samples without cavity preparation promoting the patient comfort and the healing after an
showed an odontoblast process totally in contact with the aggression.5,11,15 In this study, a conservative cavity prepara-
collagen fibrils of the extracellular matrix (Fig. 1). It was also tion was chosen, and as the samples were free of cavities the
observed the junction area of two odontoblasts bodies (Fig. 2). injury caused to the pulp tissue was due to the cavity prepare
The teeth in LG that had their class I cavities irradiated with and to the restorative process. Under such conditions, a dental
laser exhibited the odontoblast process in a bigger outline structure recovery is expected. According to Murray et al., a
contact with extracellular matrix than CG samples (Fig. 3). The remaining dentine thickness of 0.5 mm or greater is necessary
odontoblast body did not show significant morphological to avoid evidence of pulp injury.12 Constantly improved dental
changes compared with healthy samples. materials are being developed with increased mechanical and
The samples in CG, which did not receive laser irradiation, aesthetic properties, although these materials are not com-
showed spaced collagen fibrils in the extracellular matrix and pletely innocuous to dental pulp. Accorinte et al. demon-
a retracted odontoblast process (Fig. 4). It could also be strated that dental pulps covered with adhesive system were
observed in the predentine, some collagen fibrils forming inflamed or necrotic.16 Thus, the restorative process can
scattered collagen fibrils. No significant morphological differ- represent by itself an aggression to the pulp tissue.
ences were observed in the junctional area between the The sample characteristics are of great importance for this
odontoblasts comparing to healthy samples. type of study, since dentine deposition occurs during the
whole life.2 Thus, the donor’s characteristics as age, occlusal
status and historic of dental trauma can promote alterations
4. Discussion in the dentin deposition and in the pulp response. The donors
in this study presented similar ages (24 and 27 years old) and
The ultrastructural analysis at the pulp–dentine interface 28 hence the pulp tissue presents similar conditions relating to
days post-cavity preparation in teeth irradiated with low- pulp size and quantity of deposited dentin.
intensity red laser, fluence of 2 J/cm2, showed more organised The schedule of dental extraction was based on past work
collagen fibrils on predentine and less retraction of the that showed an increased dentine production in rats’ irra-
odontoblast process, compared to premolars that were diated-molars 28 days after cavity preparation compared to
902 archives of oral biology 52 (2007) 899–903

Fig. 3 – Electron micrograph of predentine region in the

pulp–dentine interface of a premolar that had cavity
prepared and did not receive laser treatment 28 days post-
Fig. 4 – Electron micrograph of predentine region in the
preparation. Note the odontoblast process (O) restrained
pulp–dentine interface of a premolar that had cavity
without filling completely the space in contact with the
prepared and received laser treatment, 28 days post-
collagen fibrils (C) of the extracellular matrix. T15,000.
preparation. Observe the odontoblast process (O) in
contact with the collagen fibrils (C) of extracellular matrix,
which are more aggregated. T22,500.
non-irradiated samples.5 Studies in literature indicate that LILT
may be used as an aide on dentine formation.5,11 In this study,
on the CG specimens, a restrained odontoblast process was the pulpal tissue.17 The results obtained in this study, confirm
observed (see Fig. 3). Although the retraction may be artifactual, these data since odontoblast damage was not observed after
all the samples, including LG specimens, received the same laser irradiation. Junctions between the odontoblast cell
processing and preparation, and they were all processed at the bodies did not present significant changes among the speci-
same time. Moreover, control group also showed collagen fibrils mens. In fact, the red laser physical properties permit a high
more randomly disperse in the extracellular matrix when penetration in hard tissue since its absorption by water and
compared to laser group (compare Figs. 3 and 4). dental mineral content is low. Beside the wavelength, it is also
It is well-known that during the healing process, the important to state the relevance of other laser parameters as
collagen fibrils initially present a random distribution. As the fluence, irradiance, frequency, etc., that can affect the
healing process continues the collagen fibrils become more outcome. In this work, the fluence used was 2 J/cm2 in a
organised, thus, the organization of the collagen fibrils can be single application, based on previous works,5,11 and also it was
used as an indicative of the healing stage. The findings of this chosen to provide a feasible application, since the process
work suggest that laser radiation may accelerate the recovery involved in dental cavity prepare and restoration is made on a
of dental tissues, since the irradiated samples presented single day procedure. According to the results, the applied
higher organization of the collagen fibrils in the extracellular parameters are appropriated to accelerate the dental structure
matrix than the non-irradiated samples. recovery. Meanwhile, the mechanisms involved remain
The wavelength of 660 nm used in this study was based on multiple and unclear leading to questions regarding reprodu-
previous works.5,11 According to Kimura et al., this wavelength cibility and effectiveness. Thus, further works are necessary to
does not affect the dentin morphologically and a fraction of clarify the exact method of action of the low-intensity laser
the incident radiation is transmitted through dentine and radiation over dental tissues. Although the mechanisms
reaches the pulp tissue. In addition, red lasers at several involved remain indistinct, this study demonstrated the
wavelengths with output power range from 30 to 60 mW do security of this therapy under the aforementioned parameters
not cause significant intrapulpal temperature rise, therefore, and the results indicated acceleration into the recovery of
these laser devices used at correct parameters do not damage dental structures after cavity preparation with a single
 archives of oral biology 52 (2007) 899–903 903

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[9] Strang R, Moseley H, Carmichael A. Soft lasers-have they a
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Acknowledgements Effect of irradiation by semiconductor laser on
responses evoked in trigeminal caudal neurons by
The authors would like to thank Dr. Rosely Cordon, for her tooth pulp stimulation. Lasers Surg Med 1993;13:
collaboration on this research, and would like to express their 605–10.
[11] Paschoud Y, Holz J. Effect du soft-laser sur la néoformation
deep gratitude to Dr. Eduardo de Bortolli Groth (in memoriam).
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