Received: 6 February 2019 Revised: 20 June 2019 Accepted: 28 June 2019

DOI: 10.1002/jbm.b.34446

ORIGINAL RESEARCH REPORT

Bioactive glass and high-intensity lasers as a promising

treatment for dentin hypersensitivity: An in vitro study

Ester M. R. Lee1 | Roger Borges2 | Juliana Marchi2 | Carlos de Paula Eduardo1 |

Márcia M. Marques3

1
Department of Restorative Dentistry, School
of Dentistry, University of Sao Paulo,
São Paulo, SP, Brazil
2
Center for Natural Sciences and Humanities,
Federal University of ABC, Santo André, SP,
Brazil
3
Post Graduation Program, School of
Dentistry, Universidade Ibirapuera, São Paulo,
SP, Brazil
Correspondence
Márcia M. Marques, Post Graduation Program,
School of Dentistry, Universidade Ibirapuera,
Av. Interlagos, 1329-4
Chácara Flora, São
Paulo, SP 04661-100, Brazil.
Email: marcia.marques@pq.cnpq.br
Funding information
CNPq, Grant/Award Numbers: 134268/2015-
6, 307874/2014-1, 130637/2016-5; FAPESP,
Grant/Award Numbers: 2016/16512-9, 2011/
19924-2
Abstract
This in vitro study aimed to analyze the physical and chemical characteristics of the
hypersensitive human dentin-like surface after application of a bioactive glass
(BG) paste (BG/Ac) irradiated or not with high-power lasers. Dentin specimens were
treated with 17% Ethylenediamine tetraacetic acid (EDTA) solution to mimic a hyper-
sensitive dentin and then submitted to neodymium: yttrium–aluminum–garnet (Nd:YAG)
laser or CO2 laser irradiation prior and after application of BG/Ac. Characterizations were
performed by using X-ray diffraction, Fourier transformed infrared spectroscopy, scanning
electron microscopy, and energy dispersive X-ray spectroscopy. The results suggested
that application of BG/Ac by itself caused some obstructions of dentinal tubules. Nd:YAG
laser irradiation reduced the opening of the dentinal tubules with no changes in the colla-
gen structure. CO2 laser irradiation caused dentin melting and resolidification along with
cracks and chemical changes in collagen fibers. However, when BG/Ac paste was irradi-
ated with lasers, a sequence of surface reactions between glass and dentin interface led
to the formation of an amorphous hydroxyapatite layer, similar to that of an inorganic
component of the normal dentin. Moreover, BG/Ac was able to prevent the formation of
cracks and degradation of collagen fibers caused by CO2 irradiation. Overall, this study
supports that application of BG/Ac paste irradiated by high-power laser could represent
an effective and long-lasting therapeutic approach for dentin hypersensitivity.

KEYWORDS
bioactive glass, Brushite, CO2 laser, dentin hypersensitivity, laser, Nd:YAG laser

1 | I N T RO D UC T I O N is a consequence of mechanical and acidic challenges to which teeth

Dentin Hypersensitivity (DH) is an increasingly frequent condition and
currently represents one of the foremost complaints of patients in
dental offices (Ladalardo et al., 2004; Orchardson & Gillam, 2006;
West, 2008). It is characterized as a transient, acute, and rapid pain
originating from a stimulus (e.g., thermal, mechanical, osmotic, and/or
chemical) on an exposed dentin surface. Dentin exposition allows the
dentinal tubule fluid to move with response to the aforementioned
stimuli, which generates DH (Brannstrom & Astrom, 1972). Thus, DH
are subject during the patient's life, causing structural losses and, con-
sequently, formation of noncarious lesions (Addy, 1992; Ling & Gillam,
1996). The treatments available for DH are based on the analgesic
effect caused either by obliteration of dentinal tubules (i.e., by using
agents like resins, glass-ionomers cements, and toothpastes con-
taining bioceramics), which avoids the movement of the dentinal fluid
inside the tubules, or by blockage of the nerve response associated
with mechanical receptors of the pulp (Canadian Advisory Board on
Dentin Hypersensitivity, 2003). However, these treatments are not

J Biomed Mater Res. 2019;1–9. wileyonlinelibrary.com/journal/jbmb © 2019 Wiley Periodicals, Inc. 1

2 LEE ET AL.
always effective for every patient(Panduric, Knezevic, Tarle, & Sutalo,
2001). Moreover, treatments using agents to obliterate dentinal
tubules may not be long lasting when teeth are exposed to erosive
and acidic challenges from daily brushing and acidic food intake
(Kerns et al., 1991; Panduric et al., 2001). Thus, the search for new
materials and treatments is still of importance. Among the proposals
for treatment of DH, there are those based on laser irradiation associ-
ated or not with application of bioactive materials to the surface of
the exposed dentin (Bakry et al., 2011; Farmakis, Beer, Kozyrakis,
Pantazis, & Moritz, 2013; Ladalardo et al., 2004; Lee, Chang, Chen,
Lan, & Lin, 2005; Lee, Tsai, Tsai, Lan, & Lin, 2005). These treatments
aim to occlude the dentinal tubules not only at the surface level, but
also from inside out by stimulating the formation of reparative dentin
(Farmakis et al., 2013; Ladalardo et al., 2004; Lee, Tsai, et al., 2005;
Lopez et al., 2017).
High-power lasers, such as carbon dioxide (CO2), neodymium:
yttrium–aluminum–garnet (Nd:YAG), erbium: yttrium–aluminum–garnet,
and erbium, chromium: yttrium–scandium–gallium–garnet, are some
types of lasers used in dentistry. Among them, Nd:YAG laser is vastly
used for treating DH because of its unique mode of action in melting
the dentin surface with subsequent resolidification, which leads to
occlusion of exposed dentinal tubules, preventing dentinal fluid move-
ment, and consequently decreasing the symptoms of DH (Farmakis
et al., 2013; Gutknecht, Moritz, Dercks, & Lampert, 1997; Naylor,
Aranha, Eduardo Cde, Arana-Chavez, & Sobral, 2006). CO2 lasers have
also been proposed as a therapy for management of DH due to the
reduction of dentinal tubules and dentin permeability (Gholami,
Fekrazad, Esmaiel-Nejad, & Kalhori, 2011; Moritz et al., 1998; Romano,
Aranha, da Silveira, Baldochi, & Eduardo Cde, 2011; Steiner-Oliveira
et al., 2006; Steiner-Oliveira, Nobre-dos-Santos, Zero, Eckert, & Hara,
2010; Steiner-Oliveira, Rodrigues, Lima, & Nobre-dos-Santos, 2008).
Bioceramics like tricalcium phosphate, hydroxyapatite, and bioac-
tive glasses (BGs) are the most applied materials for the treatment of
DH (Borges, da Silva, & Marchi, 2012; Marchi, 2016). BGs have the
ability to promote the formation of hydroxyapatite on the dentin sur-
face (Bakry et al., 2011; Borges et al., 2012; Curtis, West, & Su, 2010;
Gillam, Tang, Mordan, & Newman, 2002; Godoy, Arana-Chavez,
Nunez, & Ribeiro, 2007; Lee, Kang, Wang, Lin, & Lin, 2007). As the
hydroxyapatite layer is similar to the mineral hydroxyapatite found in
the dentin, both can be chemically bonded to each other to promote a
stronger adhesion of the glass particles to the dentin surface (Marchi,
2016). BG can be obtained through different synthesis methods, with
fusion quenching and sol–gel being the most used ones. The sol–gel
synthesis has the advantage of producing glass particles with con-
trolled morphology and size (Xia & Chang, 2007) appropriate for
occlusion of dentinal tubules (Lee et al., 2007). In addition, our
research group has already demonstrated that BG has biocompatibility
and can leach substances in aqueous substrate capable to induce cell
migration, adhesion, proliferation, and differentiation of cultured
human dental pulp stem cells (Lopez et al., 2017). Thus, BG could
induce the formation of reparative dentin to act as a biological seal of
dentin tubules. This reparative dentin is thought to have low perme-
ability, which hinders the diffusion of harmful agents from dentinal
tubules to dental pulp and reduces the movement of dentin fluid, thus
minimizing the DH. Therefore, application of BG to the surface of the
exposed dentin could represent a long-lasting treatment for DH.
Although the application of BG is a promising approach for long-
lasting treatment of DH, BG can also be lost when exposed to erosive
and acidic challenges from daily brushing and acidic food intake
(Curtis et al., 2010; Miglani, Aggarwal, & Ahuja, 2010). Therefore, it is
important to find an effective form of carrying this material to the
dentin surface and hold it there. The hypothesis of the present study
is that high-power laser irradiation could fulfill this task. Irradiation of
BG previously applied to dentin could cause physical and chemical
changes on the dentin surface, which would guarantee the durability
of BG on dentin surface long enough not only to obliterate the den-
tinal tubules, but also to allow BG to exert its bioactivity over the den-
tal pulp. For testing this hypothesis, this in vitro study analyzed the
physical and chemical characteristics of human dentin surface after
application of a synthetic BG paste irradiated or not with Nd:YAG or
CO2 lasers.
2 | MATERIALS AND METHODS
This study was approved by the Research Ethics Committee of the
University of Sao Paulo, School of Dentistry according to protocol
number CAAE#60108016.6.0000.0075.
2.1 | Preparation of dentin specimens
Forty-eight extracted noncarious third molars were cleaned and all
soft tissues around the root surfaces were removed. Then, root scal-
ing was manually performed to remove the cementum layer in order
to expose the dentin surface. The crowns were sectioned at the
enamel–cement junction by using double-faced diamond discs
(KG Sorensen Ind. e Com. Ltda, São Paulo, Brazil) at low speed and
with water cooling. The roots were then sectioned longitudinally in
the buccal-lingual direction, resulting in two dentin specimens per
tooth (n = 96). A cervical window measuring 9 mm2 was made in the
surface of each specimen. All the specimens were stored in distilled
water at 4
C, which was changed weekly until the next step.
In order to simulate a hypersensitive dentin surface, all the speci-
mens were immersed into 17% EDTA solution (Formula e Ação, São
Paulo, Brazil) for 2 min. Next, they were washed with distilled water
for 30 s to obtain a dentin surface with open dentinal tubules. These
specimens were randomly distributed into six experimental groups
(n = 16 per group) according to the treatments used and considering
laser and BG as variables studied (Table 1).
2.2 | Synthetic BG and paste preparation
BG based on the system 58SiO2–33CaO–9P2O5 was synthesized by
using the quick alkali sol–gel method, which is described elsewhere
(Borges & Marchi, 2016; Xia & Chang, 2007). The resulting powder
was mixed with a 30% H3PO4 solution (0.05 g/0.1 mL), forming a
LEE ET AL. 3

TABLE 1 Experimental groups of dentin specimens 40 kV, 40 mA, 5
< 2θ < 70
, time step of 10 s and step size of
0.02 Â
. The FTIR spectra were obtained by using a 610-IR spectrom-
Group Group Description
eter (Varian, Agilent, Palo Alto, CA) in attenuated total reflectance
1 Control (C) With no further treatment
mode from 500 to 4,000/cm. FTIR data were analyzed and deco-
2 Bioactive glass paste Treated with the BG
(BG/ac) dispersed into 30%
nvoluted by using the FityK® software (version 0.9.8).
phosphoric acid
3 Nd:YAG laser (Nd:YAG) Treated with Nd:YAG laser
3 | RESULTS
irradiation
4 BG/ac plus Nd:YAG laser Treated with the Nd:YAG
(BG/ac/Nd:YAG) laser irradiation after the
3.1 | Characterization of the dentin
application of the BG/ac
The dentin was analyzed by SEM before and after acid etching. Before
in the dentin surface
etching (Figure 1a), the dentin surface was mostly covered by smear
5 CO2 laser (CO2) Treated with CO2 laser
layer and some tubules were exposed. After acid etching (Figure 1b),
irradiation
the dentin surface became free of smear layer free. Dentinal tubules
6 BG/ac plus CO2 laser Treated with the CO2 laser
(BG/ac/ CO2) irradiation after the were opened and presented diameters greater than those before
application of the BG/ac etching. EDS spectrum (Figure 1c) showed that dentin surfaces were
in the dentin surface mostly composed of calcium, phosphor, and carbon, which are associ-
Abbreviations: BG, bioactive glass; Nd:YAG, neodymium: ated with crystals of hydroxyapatite (Ca and P) and collagen fibers (C).
yttrium–aluminum–garnet. Carbon content was also related to carbon coating.
XRD results (Figure 1d) showed a semicrystalline spectrum. The
BG/Ac paste. This paste was actively applied on the dentin surface of
amorphous contribution (expressed as a broad band between 20
and
the specimens of Group 2 (G2), Group 4 (G4), and Group 6 (G6) by
30
) was related to collagen fibers, whereas peaks are related to
using a rotating brush at low speed for 15 s. The paste remained on
hydroxyapatite crystals (ICDD-PDF 086-0740). FTIR spectra results
the surface of these specimens for 5 min. Next, the specimens of G4
(Figure 1e) were divided into two different areas, blue and red, where
and G6 were irradiated with Nd:YAG and CO2 lasers, respectively.
most of the inorganic and organic groups could be, respectively,
found. The inorganic functional groups were assigned to the mineral
2.3 | Laser treatments hydroxyapatite of dentin, where different vibration modes of PO4− 3
Specimens of Group 3 (G3) and G4 were irradiated with a Nd:YAG were identified (ν1 at 960/cm, ν4 at 1,000–1,020/cm, and bending
®
laser (Power Laser TM ST6, Lares Research , Chico, CA) operating at mode at 1,100/cm)—which are typical of apatite. In the organic func-
1064 nm and 1.5 W in a pulsed form, contact mode, repetition rate of tional groups, side chains N H and C H (1,380–1,450/cm), amide II
10 Hz and 100 mJ of energy. The irradiation was performed perpen- (1,530–1,570/cm), amide I (1,600–1,700/cm), and a collagen band
dicularly in scanning movements by using a 400-μm quartz fiber in (1,200/cm) were found, all related to collagen fibers.
which four irradiations of 15 s each were performed. Group
5 (G5) and G6 were irradiated with a continuous emission CO2 laser
3.2 | Glass characterization
(Union Medical Engineering Co., CO2 laser system, UM L-30) operat-
ing at 10,600 nm, 1 W, 125.38 W/cm2, and emitting a guide light by a Figure 1 also shows the initial characterization of BG particles. XRD
5-mW diode laser in unfocused mode at a distance of 30 mm from data (Figure 1f) showed a typical amorphous behavior characterized
the dentinal surface. Specimens were irradiated six times in 5-s bursts. by a broad peak between 20
and 30
related to the short-range peri-
The beam diameter was 0.5 mm and a 10-s interval between the irra- odicity of silicon tetrahedrons of glass structure. Small peaks were
diations was given for thermal relaxation during both procedures. also found in the spectrum, which were attributed to a crystalline cal-
cium pyrophosphate phase (Ca2P2O7 ICDD-PDF 033-0297). There-
fore, BG particles are heterogeneous, mostly containing an
2.4 | Characterization
amorphous phase and a small crystalline fraction, as already reported
The dentin, BG, and dentin surface of all experimental groups were by Hong et al. (Hong, Liu, Chen, Chen, & Jing, 2009). The FTIR spec-
characterized by using scanning electron microscopy (SEM), energy trum (Figure 1g) shows the presence of silicate and phosphate func-
dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD) analysis, tional groups, which are listed in the graph. The deconvolutions of the
and Fourier transformed infrared spectroscopy (FTIR). FTIR spectra showed peaks associated with BG structure, such as
SEM and EDS analyses were performed by using a scanning elec- SiO2 (800/cm), SiO4 (989/cm), Q0 (847/cm), Q3 (1,125/cm), Q4
tron microscope (FEI QUANTA 250, Hillsboro, OR) and an energy dis- (1,200/cm), Si O Ca (1,060/cm) ones. The phosphate band at
persive X-ray spectrometer (JSM 6010 LA Jeol, Tokyo, Japan), 1,030/cm may belong to either glassy or crystalline phosphates. The
respectively. XRDs patterns were collected with a D8 Focus diffrac- main Qn species observed were Q2 and Q3, which is in agreement with
tometer (Bruker AXS, Madison, WI) using Cukα and operating at results of the literature (Aguiar, Serra, González, & León, 2009) as the
4 LEE ET AL.
structure of BGs has these species as being the most abundant among
the other ones. SEM micrographs (Figure 1h) show that glass particles
were mostly composed of large micrometric aggregates made of smaller
particles (<2 μm), which means that glass particles were small enough
to occlude the exposed dentinal tubules. EDS analysis (Figure 1i)
showed peaks of the elements present in the glass composition, that is,
Si, Ca, P, and O. The carbon peak was associated to carbon coating.
3.3 | Characterization of the dentin in the
experimental groups
Representative SEM images of the dentin surface of all experimental
groups and EDS analysis of the groups treated with BG/Ac plus lasers
F I G U R E 1 Characterization of the
dentin with SEM micrographs before
(a) and after acid etching (b), EDS (c),
XRD diffraction (d) and FTIR spectra
(e) and characterization of BG particles
with XRD diffraction (f), FTIR spectra (g),
SEM micrograph (h) and EDS analysis (i).
BG, bioactive glass; EDS, energy
dispersive X-ray spectroscopy; FTIR,
Fourier transformed infrared
spectroscopy; SEM, scanning electron
microscopy; XRD, X-ray diffraction
are presented in Figure 2. G1 had a clean dentin surface with open
tubules (Figure 2a). In G3, the dentin irradiated with Nd:YAG laser
(Figure 2e) presented dentinal tubules of smaller diameters in compar-
ison with G1, probably due to a process of melting and resolidification
of the same surface. In G5, the dentin surface irradiated with CO2
laser was melted, exhibiting areas of fusion and resolidification in the
dentin substrate. In areas with no melting, CO2 irradiation resulted in
cracking and pores of varied sizes in the intertubular dentin
(Figure 2j). The peritubular dentin seemed melted, showing tubules
with small openings and wavy aspect (Figure 2e).
G2 showed dentin mostly covered by crystalline-like structures
arranged in clusters (Figure 2b). At higher magnification, it was possi-
ble to observe smaller particles of BG in the dentin (i.e., underneath
LEE ET AL. 5

F I G U R E 2 Representative SEM images of the dentin surface in all experimental groups and EDS analysis of the groups treated with
BG/ac. (a) G1 (control) shows a clean dentin surface with open tubules; (b–d) SEM images showing dentin mostly covered by crystalline-like
structures in G2 (BG/ac), including EDS (b,c) and EDS analysis (d); (e) dentin irradiated with Nd:YAG laser (G3) showing a clean surface with
peritubular dentin with tubules with small openings and wavy aspect (arrows); (f–h) G4 (BG/ac + Nd:YAG laser) showing BG/ac on the surface of the
dentin surface as small particles closely attached to the intertubular dentin (f) and entering the dentin tubules (g) and (h) EDS analysis; (i,j) typical
dentin surface irradiated with CO2 laser in G5. In areas with no melting, the CO2 irradiation resulted in cracking and pores (arrows) in the
intertubular dentin (j); (k–m) CO2 irradiation of the dentin previously treated with BG/ac (G6). The surface was coated with very small particles of
BG/ac homogeneously distributed in the intertubular dentin (k). At higher magnification, it was possible to observe these particles inside the dentinal
tubules (l); M shows EDS analysis. EDS graphs show Si peaks in all obtained spectra. Peaks of ca, P, and O were observed in all spectra. BG, bioactive
glass; EDS, energy dispersive X-ray spectroscopy; Nd:YAG, neodymium: Yttrium–aluminum–garnet; SEM, scanning electron microscopy

these clusters) homogeneously distributed in the intertubular dentin,
partially occluding the dentinal tubules (Figure 2c). This partial occlu-
sion was also noted in those groups treated with BG/Ac plus Nd:YAG
laser (G4, Figure 2f,g) or CO2 laser (G6, Figure 2k,l). Additionally, G4
had dentinal tubules with diameters greater than those of the dentin
surface directly irradiated with Nd:YAG laser (G3), whereas dentin
specimens in G6 showed a surface with dentinal tubules smaller than
those of the control group (G1). The melting and cracking observed in
G5 were not present in the specimens of G6.
The EDS spectra of G2 (Figure 2d), G4 (Figure 2h), and G6
(Figure 2m) showed Si-related peaks. Peaks of Ca, P, and O were
observed in all analyzed spectra.
Figure 3 shows XRD (Figures 3a,c,e) and FTIR (Figures 3b,d,f) results
for the pairs G1–G2, G3–G4, and G5–G6. Figure 3a shows the compari-
son of the XRD patterns between dentin, BG, BG/Ac, and BG/Ac. The
XRD patterns of dentin surface and BG were previously detailed. As BG
was mixed with phosphoric acid (BG/Ac), a hydrated calcium mono-
hydrogen phosphate (Ca(H2PO4)2H2O, ICDD-PDF 009-0347) was
nucleated as a result of chemical reaction between glass surface and
phosphoric acid, as already reported by Patel et al. (2012). When the
paste was applied to the dentin surface, the Ca(H2PO4)2H2O phase of
BG was converted into brushite (CaHPO42H2O, ICDD-PDF 009-0077).
Similar comparisons were made between G3 and G4 (Figure 3c)
and between G5 and G6 (Figure 3e). In either results, when dentin
was irradiated with Nd:YAG (Figure 3c) or CO2 (Figure 3e) laser, no
structural changes were observed in the inorganic fraction of the den-
tin. The diffraction patterns were similar to those of the untreated
dentin, and the peaks were assigned to hydroxyapatite. When lasers
were applied to the brushite layer, which was formed when the paste
of BG was spread on the dentin surface, a similar spectrum was
observed in the untreated dentin of the specimens.
The deconvolution of the FTIR spectra for the dentin specimens
irradiated with Nd:YAG and CO2 lasers, G3 (Figure 3d) and G5
(Figure 3f), respectively, showed no structural changes in the inor-
ganic phase. In G4 and G6, peaks associated with BG were observed,
such as SiO2 (800/cm), SiO4 (989/cm), Q0 (847/cm), Q3 (1,125/cm),
Q4 (1,200/cm), and Si O Ca (1,060/cm) structures. In the functional
groups, PO4 (1,005/cm), P O P (906/cm), C O and Q0 (875/cm),
6 LEE ET AL.

F I G U R E 3 XRD diffraction (a,c,e) and FTIR (b,d,f) spectra of experimental groups: G1 and G2 (a,b), G3 and G4 (c,d), and G5 and G6 (e,f). FTIR,
Fourier transformed infrared spectroscopy; XRD, X-ray diffraction

and PO4−3 (955/cm) were found to be associated regarding all spectra Si O Si bonds and also P O P bonds occurs when the solution con-
(Figure 3). taining glass precursors is alkalinized, leading to the formation of
SiO2 P2O5 particles. When drying the solution by lyophilization, the
ionic calcium crystallizes into calcium nitrate (its precursor) around the
4 | DISCUSSION
particle of SiO2 P2O5. Calcium nitrate thermally degrades when sub-
mitted to calcination, releasing NO2, while calcium ions diffuse from the
BGs, as well as laser irradiation, have been proposed as treatment for
edges to the center of the primary particles. Considering that phospho-
DH (Bakry et al., 2011; Farmakis et al., 2013; Ladalardo et al., 2004;
rus and calcium are elements with high chemical affinity, there is the
Lee, Chang, et al., 2005; Lee, Tsai, et al., 2005). In a previous study
formation of a calcium phosphate phase near to the surface of the glass
(Lopez et al., 2017), we showed that BG had a bioactivity toward oste-
particles, where there is a higher concentration of calcium ions. A
ogenic differentiation of stem cells derived from human exfoliated
model of this mechanism is shown in Figure 4.
deciduous teeth. This would indicate that the application of this BG to
From the obtained results, it was verified that the formation of
the surface of a hypersensitive dentin could induce the formation of
heterogeneous particles was beneficial for facilitating surface reac-
reparative dentin. However, this effect could be improved if the glass
tions between BG and phosphoric acid. In these reactions, the
particles could react with the dentin and hold in place long enough
Ca2P2O7 phase resulting from BG particles is dissolved in the phos-
not only for obliteration of the dentin surface tubules, but also for BG
phoric acid solution and then precipitated as Ca(H2PO4)2H2O
to exert its bioactivity to the dental pulp. With the hypothesis that
(Equation 1). In G2, the FTIR spectra shows peaks either from the
high-power laser irradiation could fulfill this task, the aim of this study
amorphous phase of BG or from the crystalline phase of
was to analyze the physical and chemical characteristics of the dentin
Ca(H2PO4)2H2O, as can be seen in Figure 4b. In addition, a band at
surface after application of BG/Ac irradiated or not with Nd:YAG laser
or CO2 laser. Firstly, we have addressed the characterization of glass and 889/cm is associated with HPO4−2 groups, which in turn is related to

dentin as well as the reactions between glass, lasers, and dentin surface. the excess of phosphoric acid which had not been reacted.

It was noted that the glass particles were heterogeneous, consisting

of a glassy and a crystalline phase. The mechanism in which a glass is Ca2ðglass
+ 2−
Þ + 2HðaqÞ + 2HPO4 ðaqÞ + H2 O ! CaðH2 PO4 Þ2 H2 O:
+
ð1Þ
processed through sol–gel method supports the understanding of the
calcium pyrophosphate nucleation along the synthesis. According to When the BG/Ac is dispersed on the dentin surface, new phase
Lin, Ionescu, Pike, Smith, and Jones (2009), a condensation reaction of changes were observed. According to the model proposed on
LEE ET AL.
FIGURE 4 Illustrative representation of the mechanism of formation of BG particles. BG, bioactive glass
Equation 2, the unreacted HPO4−2 anions partially solubilize superficial
2+
hydroxyapatite from dentin surface, leaching Ca that, in turn, react
with the Ca(H2PO4)2H2O phase, forming brushite. The formation of
this phase was observed in the works done by Bakry et al. (2011),
Bakry, Takahashi, Otsuki, & Tagami, 2013), who reported that such
chemical reaction can be formed due to the abundant calcium concen-
tration from the BG and dentin, and it is precipitated due to the low
pH of the BG/Ac paste.
CaðH2 PO4 Þ2 H2 O + Ca2ðdentin
+
Þ ! 2CaHPO4 2H2 O + H : +
ð2Þ
A process of melting and resolidification was noted only when Nd:
YAG (G3) or CO2 (G5) lasers were applied to the dentin, but which
was much more intense in G5, where the organic phase of dentin was
clearly damaged. On the other hand, when the irradiation occurred on
the dentin surface coated with BG/Ac, this collagen degradation was
no longer observed. Similar results were reported by Bakry et al.
(2011), 2013) who reported not only the formation of a brushite
®
phase when a mixture of NovaMin and phosphoric acid was depos-
ited on the dentin surface, but also the formation of a monetite phase
when the acid paste was irradiated with CO2 laser. This finding indi-
cates that the crystalline phase of brushite can absorb the photon
energy from CO2 lasers, which leads to dehydration reactions as mon-
etite is an anhydrous phase of brushite. Although our results showed
that the brushite phase formed in our acid paste was not transformed
into monetite instead of hydroxyapatite, the results of both studies
are consistent. Bakry et al. used a laser of 0.5 W, whereas we have
7
used one of 1 W, meaning that the energy delivered per unit of area
onto the dentin surface was higher. Consequently, it is reasonable to
assume that monetite might be an intermediate reaction product
between brushite degradation and hydroxyapatite formation as a
higher energy could lead to further dehydration.
According to the Equation 3 proposed by Lin, Lee, Lin, Kok, and
Lan (2001), the brushite phase can be also transformed into hydroxy-
apatite and the condition for this transformation is an excess of cal-
cium in the medium, which could come from the glass or dentin.
6CaHPO4  2H2 O + ðx – 10Þ Ca2 + ! Ca10 – x ðHPO4 Þx ðPO4Þ6 – x
ð3Þ
ðOHÞ2 – x ð8 – 2xÞH + ðx + 10Þ H2 O:
Interestingly, when comparing the PO4 band (1,005/cm) of the
dentin before and after the interaction with either Nd:YAG or CO2
lasers in the presence of BG, it was observed that there was a shift to
higher wave numbers (i.e., 1,008 and 1,009/cm). This shift may be
associated with a possible formation of Si O P bonds, which would
ensure a greater adhesion of the BG on the surface of the dentin as
they would be chemically bonded to each other. A study carried out
by Bilandzic et al. (2017) showed that BG ceramics can be chemically
bonded to bovine enamel when CO2 laser is applied. These findings
indicated that chemical bonds between dentin and BGs might not
have been excluded.
For the very first time, we have reported new insights on an ele-
gant method using laser irradiation plus BG and phosphoric acid to
guide the formation of an inorganic layer very similar chemically to
8 LEE ET AL.
the hydroxyapatite of the human dentin. As suggested in our prior
study (Lopez et al., 2017), it is expected that the reparative dentin will
be formed through its bioactivity when BG remains inside the dentinal
tubules, thus completing the desensitization process by means of
physiological occlusion of the dentinal tubules.
5 | CO NC LUSIO NS
This in vitro study has shown that BG with 30% phosphoric acid can
promote obliteration of the exposed dentinal tubules and the irradia-
tion of this material with either Nd:YAG or CO2 laser improves occlu-
sion. Moreover, new mechanisms were proposed about changes in
the dentin surface induced by application of this BG/Ac, which guided
the formation of amorphous hydroxyapatite when submitted to high-
power laser irradiation.
Based on the results of the present study, one can infer that high-
power laser irradiation of the BG acidic paste present on the human
dentin surface could represent an effective and long-lasting therapeu-
tic approach for DH.
ACKNOWLEDGMENTS
The authors would like to acknowledge the UFABC/CEM facilities
(i.e., SEM, XRD, and FTIR) and governmental grant agencies CNPq
(#134268/2015-6) and FAPESP (#2016/16512-9 and
#2011/19924-2) for the financial support to this project. M.M.M. is
supported by CNPq #307874/2014-1 and R.B. by CNPq #
130637/2016-5. We would also like to thank Elisângela Chinen for
SEM technical support during preparation of the specimens and Telma
Zambanini for SEM analysis support.
ORCID
Márcia M. Marques https://orcid.org/0000-0002-9398-1252
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How to cite this article: Lee EMR, Borges R, Marchi J, de
Paula Eduardo C, Marques MM. Bioactive glass and high-
intensity lasers as a promising treatment for dentin
hypersensitivity: An in vitro study. J Biomed Mater Res. 2019;
1–9. https://doi.org/10.1002/jbm.b.34446