J Clin Exp Dent. 2020;12(3):e291-9.

Remineralizing ability of bioactive cements

Journal section: Biomaterials and Bioengineering in Dentistry doi:10.4317/jced.56500

Publication Types: Research https://doi.org/10.4317/jced.56500

Comparative evaluation of bioactive cements

on biomimetic remineralization of dentin

Nazanin Daneshpoor 1, Leila Pishevar 2

1
Restorative dentistry specialist
2
Department of operative dentistry, faculty of dentistry, Isfahan (khorasgan) branch, Islamic Azad university, Isfahan, Iran

Correspondence:
Department of operative dentistry
faculty of dentistry, Isfahan (khorasgan) branch
Islamic Azad university, Isfahan, Iran
[email protected]

Daneshpoor N, Pishevar L. Comparative evaluation of bioactive cements on

Received: 29/10/2019
Accepted: 08/01/2020
biomimetic remineralization of dentin. J Clin Exp Dent. 2020;12(3):e291-9.

Article Number: 56500 http://www.medicinaoral.com/odo/indice.htm

© Medicina Oral S. L. C.I.F. B 96689336 - eISSN: 1989-5488
eMail: [email protected]
Indexed in:
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DOI® System

Abstract
Background: This study was designed to quantitatively compare the presence of apatite peaks on demineralized
dentin to induced bio-mimetically by bioactive commercial materials.
Material and Methods: Dentin slice specimens (n=6) were prepared and demineralized by by17% EDTA for 2
hours. Each disks materials (Theracal, Biodentine, CPP-ACP) were freshly prepared and was maintained in close
contact with a demineralized dentin specimen immersed in PBS solution during one week. To evaluation of bioac-
tivity, cements disks (n=6) were prepared from each material and immersed in PBS solution for one week. The
bioactivity and remineralization ability was evaluated using FTIR spectroscopy and Scanning Electron Microsco-
py. The Ca/P ratio of the surface of dentin and cements disks were compared with one -way ANOVA, independent
T test and Duncan test (α=0.05).
Results: Ca/P weight ratio of Biodentine (187.5) was significantly higher than Theracal (10.10) and Theracal higher
than CPP-ACP (0.37) (P=0.008). Demineralized dentin in contact with Test materials, indicated Ca and P peak after
7 days, but not showed statistically differences between the groups (P=0.08).
Conclusions: The outcome revealed that bioactive cements and CPP-ACP had bioactivity capability during one
week. Biodentine had higher bioactivity between others. Demineralized dentin could be remineralized with bioac-
tive materials.

Key words: Bioactive, cements, fourier transform infrared spectroscopy, scanning electron microscopy, tooth
remineralization.

Introduction matrix, i.e. the collagen fibers, therefore cause dental

Either organic or inorganic constituent of dentin alter caries (1). Contemporary treatment of carious lesions
from bacterial by-products (organic acids) in the bio- is to remove only the infected external dentin, although
film. These acids lead to dentin demineralization sub- the affected internal dentin that can be remineralized is
sequently loss of mineral crystals, and also alter organic preserved (2).

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Today, there is a challenge to remineralize remaining together. The null hypothesis is that there are no diffe-
demineralized dentin in deep caries. Dentin reminera- rences between bioactivity of test materials and there are
lization is more difficult than enamel remineralization no differences between them in terms of induction of
that is due to excess presence of dentin organic matrix. remineralization within demineralized dentin.
Most conventional carious dentin remineralzation agent
involves the use of calcium and phosphate ions solution Material and Methods
with different concentration of fluoride. Well known that This study gathered the number 23810201952008 as
common remineralization process occurs by epitaxial approved by the medical ethics committee of Isfahan
growth of residual apatite crystals in partially demine- Azad University.
ralized dentin. It means that there is no remineralzation, -Materials: Three commercial materials (CPP-ACP, Bio-
if there are a little residual crystals or none (3). Biomi- dentine, Theracal) were tested in this study. For the pre-
metic treatment of affected caries dentin, involve guided paration of each disk material, Teflon molds (5 mm dia-
remineralization followed by or mimicry the physiolo- meter and 2 mm thickness) were used. Theracal (Bisco,
gic mechanism of tissue mineralization. This strategy is Schaumburg, IL, USA, LOT 1700002279) was injected
independent of growth of remaining apatite crystals so to the mold, slightly pressed with a sliding glass and li-
partially and well-demineralized dentin can be success- ght cured with LED system (1200 mW/cm2, Demetron
fully remineralized. Bioactive glass, poly electrolytes, Demi, Kerr, USA) for 20 secs from each side. Each The-
amorphous calcium phosphate, and calcium silicate ce- racal disk had 0.13 gr weight. Biodentine (Septodont,
ments are bioactive materials that has been investigated Saint Maur-des-Fossec, France, LOT B19093) was pre-
to bio-mimetically remineralize the dentin (4). pared following manufacturer’s instructions (five drops
Amorphous calcium phosphate is a precursor of hydrox- of liquid added to powder and triturate in amalgamator
yapatite that lead to precipitate calcium phosphate ions, for 30 seconds) then condensed the cement in mold with
as a final stable by-product (5). The casein phospho-pep- a plastic spatula (provided with Biodentine, Septodont)
tides are resulted from tryptic digest of casein (6) and and left to set until 12 min. each Biodentine disk had
have the ability to stabilized calcium and phosphate ions 0.15 gr weight. The construction of CPP-ACP disk, 5%
through releasing small peptide sequences (CPP) resul- weight ratio of CPP-ACP (tooth mousse, GC corpora-
ted from partial enzymatic breakdown that led to remi- tion, Tokyo, Japan, LOT 160804S) added to the formu-
neralization (7,8). Casein phospho-peptide have a high lation conventional glass ionomer luting cement (Fuji
binding affinity for apatite (9). Calcium silicate cements VII, GC corporation, Tokyo, Japan, LOT 160527A) in
are hydrophilic materials that tolerate humidity and set order to set this material as a disk like specimen and
in biologic solutions (blood, plasma, saliva or dentinal evaluating the bioactivity of material (12). This prepa-
fluid). Wetting of tri-calcium silicate cements, cause to red paste was placed in the mold and let to set. Each disk
produce calcium silicate hydrate gel (C-S-H gel) and had a 0.11 gr weight.
calcium hydroxide. Elongated setting time and the need -Bio- property analysis:
to cover with other materials to let them fix are major Apatite deposition on the surface of material disks (Apa-
deficit of the use of MTA like materials, as a pulp cap tite forming ability test): the apatite forming ability (bio-
material (10). Some tri-calcium silicate base material inter activity/ bioactivity) was studied by evaluation of
has introduced with shorter setting time as Biodentine. apatite formation on disk material in the presence of a
Theracal, is a light cure resin modified calcium silicate simulated body fluid (SBF) that in this study the PBS
cement as base and liner that designated for direct and (Phosphate Buffer Saline) was used. PBS is a physiolo-
indirect pulp cap material beneath the resin composite gic saline solution with 7.4 pH, Ca and Mg free, contai-
restorations with the goal of bond between different ma- ning some salts (4.18 K+, 152.9 Na+, 139.5 Cl- and 9.56
terial layers and lower micro-leakage. It has shown that Po4-3 (1.5 H2PO4- and 8.06 HPO42-)). Buffering acid-base
release higher amount of calcium comparison to MTA was added to preserve the pH. Salt concentration of this
and Dycal, therefore it has the ability to alkalizing the solution was considered as body fluids (isotonic) like
surrounding environment (11). plasma (13).
The purpose of this study is to gain insight in calcium After preparation of each disk material (n=6 in each group),
silicate cements (Biodentine and Theracal) and CPP- soaked them in PBS solution separately and kept in plastic
ACP with each other in terms of bioactivity and remine- container (3cm height and 4 cm diameter) at 37°C until 7
ralizing ability of demineralized dentin within the short days (a PBS/cement ratio of 17 ml/gr was used). Then sur-
period (7 days’ immersion time) in phosphate-contained face chemistry (surface component and elemental distribu-
solution (human like situation). Bioactivity and remine- tion of phases) was studied in dry condition by SEM-EDX
ralization ability of this three-mentioned materials have and ATR-FTIR Spectroscopy methods.
been separately analyzed before, but the novelty of this Dentin Remineralization Test (DRT): Remineraliza-
study is the quantitative comparison of these properties tion property (Bio-remineralizaton) was studied by the

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protocol (explains as following) of ability to induce the neity, five IR spectra were done at five point of different
formation of apatite on previously demineralized dentin surface of each specimen.
(13). -Statistical analysis
Dentinal samples were collected from healthy human Bioactivity and remineralizing ability data were statisti-
third Molar. The teeth were kept in 0.5% thymol in 4°c cally analyzed by one -way ANOVA Test, independent T
for a maximum 3 months’ duration. Dentinal slices with –Test and Duncan Test with 22 version of SPSS software
1± 0.1 mm thickness were prepared by an automatic cu- package.
tting-instrument (CNC cutting machine, Tabriz, Iran).
These sections were parallel to CEJ of tooth and done Results
by a new diamond-cutting disk with cooling agent. All -Apatite deposition on the surface of material disks:
samples were checked in terms of any defects like crack SEM-EDX analysis: freshly prepared material SEM
or hole or presence of enamel or pulp tissue by means of analysis showed the presence of constituent elements.
optical microscopy. Therefore, the samples were com- After 7 days’ material soaked in PBS, SEM revealed the
pletely covered by smear layer and kept in deionized appearance of fine spheres on the surface of Biodentine
water, in refrigerator before use. The surface of dentinal and Theracal disks and in the lesser amount on CPP-ACP
slices was evaluating by EDX and FTIR. Then putted disks. In addition, Ca and P peak of the upper surface was
the samples in 15 ml 17% EDTA (Master dent, Dento- a sign of the presence of Ca-P deposits. The decrease in Si
nics Inc., USA, LOT 9515) for 2 h (at room tempera- peak could suggest a major gathering of this element in the
ture) to demineralized the samples (14). Then washed subsurface of CSH (Calcium silicate hydrate) phase and
them with deionized water several time and randomly releasing to the solution. T independent test showed that:
categorized to four groups (n=6 in each group). Disks 1) the mean P peak in three materials were significantly
of set material (CPP-ACP, Biodentine, Theracal) were higher after 7 –days soaking time than fresh materials. 2)
prepared by Teflon mold (5ml diameter and 2mm thick- The mean Ca peak in three materials was not significantly
ness). Each disk of set material was maintained in close altered after 7 –days soaking time. 3) The average of Ca/P
contact with demineralized dentin slices using an ortho- weight ratio in Biodentine group was significantly higher
dontic elastic band and soaked in 15ml PBS solution in than fresh material after 7 days soaking time (P<0.05) but
plastic container and in 37°C for 7 days. This technique in CPP-ACP and Theracal groups after 7 days was less
allowed the dentinal slices readily away from disk ma- than fresh ones (P<0.05) (Table 1). The average of P peak
terial after spent soaking time. One group of deminerali- in fresh and 7days CPP-ACP groups were significantly
zed dentin was maintained in PBS without any treatment higher than other groups (P<0.05).
as a control group. After separation, slices were rinsed FTIR Spectroscopy analysis: Figure1 shows the IR
with deionized water, dried with ethanol, and analyzed. spectra recorded from the surface of fresh material and
-Measurement 7 days’ disks in PBS. Freshly prepared Biodentine and
SEM-EDX: samples were analyzed with environmental Theracal disks revealed bands related to belite, C-S-H
Scanning Electron Microscopy connected to a secondary and commencement of portlandite and anhydrite or gyp-
electron detection for Energy Dispersive Xray analysis sum (calcium sulfate). The elite band is not seen. In fresh
EDX [FIE ESEM, Quanta,200, USA] using an accele- CPP-ACP, bands due to the reaction of carboxylic acid
rating voltage 25 Kv. Elemental analysis (weight% and with Al, Ca and formation of carboxylate salts was seen
atomic %) of samples was done by using ZAF correction and P band due to phosphate of casein phospho-peptide
method. EDX was performed on dry material disk surfa- amorphous calcium phosphate added to Glass ionomer
ce and dentinal slices. Dental samples need to be coated was seen. After 7 days on surface of Biodentine and The-
with gold for subtle discovery. Both surface of dentinal racal disks, band due to carbonate ion characteristics in
slices was analyzed (surface in contact with material [or the form of different chemical phase (calcite, aragonite)
upper] and opposite [or free] surface). and type B apatite carbonate (indeed the common Ca-P
ATR-FTIR: Infrared spectra were recorded by using band) appeared. No band due to primary cement com-
FTIR Spectroscopy (BURKER FT-IR, TENSOR27) ponent was seen that suggested apatite formation layer
equipped with attenuated total reflectance (ATR) acces- that has enough thickness (2µm) to cover the surface of
sory and DTGS detector for analysis of phase composi- cement (average penetration depth of IR spectra with the
tion and crystal structure. The spectral resolution was 4 use of diamond crystal in ATR technique is 2 µm). At the
cm-1 and 64 the number of scan for each spectrum in the end of immersion period in CPP-ACP group: 1- P-peak
region of 400-4000 cm-1. The diameter of ATR acces- relative intensity increase., 2- band due to carboxylate
sory was 2 mm diameter and the IR penetration power salt was seen meaning thin apatite layer formed on ce-
was about 2 µm. due to high absorption of IR spectra in ment surface so cement component is still recognizable.,
water, samples were dried with desiccator before spec- 3- commencement of small C=O peak showed the bioac-
troscopy. To minimize the problems due to inhomoge- tivity and little amount of apatite formation.

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Table 1: The average of Ca/P weight ratio, Calcium and phosphor wt % on test material
disks separated by time analyzed by EDX.
Fresh 7 days
Group variable P-value
Average Average
Ca/P 73.09 (4.11) 187.51 (70.18) 0.01
Ca 41.9 (1.48) 43.65 (2.25) 0.14
Biodentine
P 0.57 (0.04) 1.24 (0.71) 0.04
Ca/P 0.71 (0.08) 0.37 (0.12) 0.001>
CPP-ACP Ca 1.62 (0.2) 1.46 (0.25) 0.26
P 2.34 (0.56) 4.14 (0.69) 0.001
Ca/P 31.88 (7.99) 10.10 (3.02) 0.001>
Theracal Ca 20.29 (0.6) 19.55 (1.23) 0.21
P 0.66 (0.13) 2.07 (0.59) 0.001>

Fig. 1: FTIR analysis of fresh (A) and 7 days (B) Biodentine disks, fresh (C) and 7 days (D) CPP-ACP disks and fresh (E) and 7 days (F) Thera-
cal disks.

FTIR analysis proved: 1- presence of carbonate apatite demineralization treatment (17% EDTA, 2h) is effective
on the surface of cement after 7 days of being soaked in and can remove the mineralized phase. Indeed, P & Ca
PBS. 2- More prominent apatite phase band (a relative peak was removed and only water and collagen/protein
increase of phosphate and carbonate -band peak intensi- matrix were left on the surface.
ty) after 7 days in Biodentine. 3- Relative increase of P Demineralized dentin, without any treatment, was
peak on the surface of CPP-ACP after 7 days. soaked in the PBS solution for 7 days to ensure that no
-Dentin remineralization test: remineralization occurred during soaking time. Data re-
SEM-EDX analysis: There are some Ca, P, C and O peak vealed that no significant increase in Ca-P peak or re-
in the intact dentin surface. Results confirmed that the mineralization occurred. One-way ANOVA showed that

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the average amount of Ca, P & Ca/P ratio was different According to Table 3, independent t-test showed that the
(P<0.05) between 3 groups (intact dentin, demineralized average amount of Ca, P and Ca/P ratio in 3 groups was
dentin, control group). Duncan test showed that Ca, P significantly higher than the opposite surface (un-trea-
& Ca/P ratio in the intact group was significantly higher ted surface). According to Table 4, one-way ANOVA
than two other groups, but between the two, there are no showed that the average amount of Ca, P & Ca/P ratio in
differences (Table 2). the opposite surface between 3 groups have no signifi-
Demineralized dentin in contact with Biodentine, CPP- cant differences (P>0.05). Ca/P ratio on the Biodentine,
ACP, Theracal after 7 days of being soaked in PBS: ac- Theracal, and CPP-ACP-treated side was 2.87, 2.18, and
cording to EDX, Ca and P peak found on the surface, 2.5 respectively that there are no statically significant di-
it means that remineralization occurred. Another finding fferences between them (P>0.05).
in SEM images was the presence of apatite deposition FTIR analysis: finding obtained from FTIR spectrosco-
in forms of fine intra tubular spheres on the treated side py revealed that EDTA was able to remove the mineral
of dentin. phase from the region of 1180-885 cm-1. In fact, the IR

Table 2: Average of calcium, phosphor and Ca/P wt % on the surface of dentin slices.
Variables Intact dentin Demineralized dentin Demineralized dentin 7 days in P-value
PBS (as control group)
Average Average Average
Ca/P 2.41 (0.05) 1.9 (0.13) 1.68 (0.4) <0.001
Ca 39.56 (1.52) 2.34 (0.46) 3.09 (0.53) <0.001
P 16.44 (0.61) 1.29 (0.19) 1.93 (0.57) 0.001

Table 3: Average amount of Ca, P and Ca/P wt ratio in treated and opposite side separated bygroups.
Variable Treated side Opposite side
Group P-value
Average Average
Ca 21.82 (9.97) 2.14 (0.71) 0.001
CPP-ACP
P 8.37 (3.80) 1.06 (0.31) 0.001
Ca/P 2.55 (0.45) 2 (0.23) 0.02
Ca 13.90 (9.21) 2.55 (0.85) 0.01
Biodentine
P 4.71 (3.15) 1.31 (0.39) 0.02
Ca/P 2.87 (0.54) 1.96 (0.32) 0.005
Ca 17.78 (9.82) 2.81 (0.27) 0.004
Theracal
P 7.95 (3.58) 1.63 (0.37) 0.002
Ca/P 2.18 (0.44) 1.79 (0.41) 0.04

Table 4: Average amount of Ca, P and Ca/P weight ratio in three groups by treatment situation.
CPP-ACP Biodentine Theracal
Treatment Variable Average Average Average P-value
Ca 21.82 (9.97) 13.90 (9.21) 17.78 (9.81) 0.39
Treated P 8.37 (3.80) 4.71 (3.15) 7.95 (3.58) 0.18
Ca/P 2.55 (0.45) 2.87 (0.54) 2.81 (0.44) 0.08
Ca 2.14 (0.71) 2.55 (0.85) 2.81 (0.27) 0.24
Untreated P 1.06 (0.31) 1.31 (0.39) 1.63 (0.37) 0.14
Ca/P 2 (0.23) 1.96 (0.32) 1.79 (0.41) 0.51

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spectra recorded after demineralization only showed co- bonate and phosphate apatite band with different peak
llagen (amide band) in 1700-1100 cm-1 region. However, intensity on the surface of Biodentine, Theracal and
spectral characteristic due to apatite was not showed. CPP-ACP confirm the apatite formation and bioactivity.
Demineralized dentin after contact with experimental The intensity of these peaks relatively in Biodentine was
material for 7 days in PBS solution: dentin specimen higher than Theracal and Theracal was higher than CPP-
with different spectral was remineralized. In the IR ACP. Therefore, the null hypothesis (there are no di-
spectrum related to Biodentine and Theracal, carbonate fferences between bioactivity of materials) was rejected.
and phosphate apatite formed and signed in the presence Differences in composition could affect the amount and
of bands in about 781, 1020, 1409 cm-1. In CPP-ACP, rate of apatite deposition. The major constituent of Bio-
phosphate band related to the phase appeared in 1450 dentine (80.1%) is tri-calcium silicate (21). There is also
and band due to apatite revealed in 611 and 1028 cm-1. calcium chloride as an accelerator. On the other hand,
Theracal is consist of 45% Portland cement type III, as
Discussion a source of Ca ions (22). Both materials have hydration
A bioactive material provokes a positive response from chemical setting reaction, Biodentine hydration process
the host. This material should be able to extract a biolo- is more elongated, but there is an uncompleted hydra-
gic response in the interface and induce the formation of tion reaction in Theracal because of light curing and fast
a band between material and tissue. Bioactivity concept polymerization (21). CPP-ACP added to GI, polymeri-
(apatite-forming ability) has a close relation with bio in- zed into the composition, may cause a fewer amount of
teractivity i.e. ability to exchange data with the biologic Ca ions release.
system. This means that bioactive materials chemically All three materials after 7 days’ immersion time, show
react with body fluids in terms of tissue repair. So, the an increase in P peak, however, this peak in CPP-ACP
material ability to induce apatite formation on demine- was more both in freshly prepared and mainly after
ralized dentin (remineralization ability) is strongly re- 7 days of storage (Table 1). This could be due to the
lated to its bio interactivity and bioactivity (15). These constituent component, calcium phosphate and phos-
materials are able to form apatite on their surface in a phate absorption from PBS solution. In CPP-ACP and
short induction time (13). Different methods have been Theracal groups, the average Ca/P ratio in 7 days was
used to analyzing the efficacy of the remineralization lower than fresh material (<0.05). This could be due to
process and bioactivity in dentinal tissue. Evaluating the the formation of apatite and phosphate absorption from
methods can provide quantitative and qualitative data. PBS solution than Biodentine. The storage period did
Recent studies have evaluated the entrance of minerals not have a significant effect on remineralization efficacy,
to dentin through indirect qualitative analysis like Pola- this early remineralization was done in one week and
rized Light Microscopy (16). Semi- quantitative analysis then progressed slowly (23). Apatite formation could be
like Transverse Microradiography (17) and Transmis- due to Ca release from Ca silicate cement under a wet
sion Electron Microscopy (TEM) (18) and Spectrosco- condition and Ca-P been released from CPP-ACP. Du-
pic analysis like Raman and FTIR (19,20). However, ring soaking in fluids, Biodentine (like other calcium si-
each method has some limitations. IR and Raman are licate cement) immediately release Ca ions and make an
non- destructive novel techniques in the assessment of alkaline pH on the outer surface that leads to nucleation
the vibrational mode of silicate, carbonate, hydroxyl, and crystallization of hydroxyapatite on cement surface
phosphate. In addition, they determine the remineraliza- (21). Ca ions react with phosphate the group in PBS so-
tion effectiveness and the extent of the remineralization lution and deposit in forms of Ca-P. Higher bioactivity
process simultaneously. IR Spectroscopy is more sensi- of Biodentine could be due to higher alite/belite ratio.
tive in the determination of CSH and portlandite phase Indeed, alite release more calcium hydroxide than belite.
in the presence of bismuth oxide than Raman Spectros- This study shows that tested materials in contact with
copy. On the other hand, elite and belite band in FTIR is demineralized surface had induced a significant remine-
more defined. EDX is a subtle and repatriation analysis ralization after 7 days, all three materials on deminerali-
method in studying calcium silicate cement and bioac- zed surface revealed Ca-P peak that is confirmed by an
tive calcium phosphate materials that has an ability to EDX analysis. However, between three groups there are
detect constituent elements and components. The use of no differences. So, the second hypothesis is accepted.
multiple analysis could get the obtained data closer to On demineralized dentin surface in contact with Bio-
reality. dentine and Theracal, carbonate and phosphate IR band
This study clearly shows that the Calcium silicate ce- appeared that confirmed the dentin remineralization.
ment and CPP-ACP has bioactivity property, due to re- The appearance of IR band related to phosphate band in
sults from SEM- EDX (finding of fine spherules on the about 1000 cm-1 region, showed new apatite formation
surface of materials with high Ca/P weight ratio. Moreo- that approximately matched with normal dentin spec-
ver, according to result from FTIR, the presence of car- trum.

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In Gandolfi study, the SEM of the fresh Biodentine sur- months. In the present study SEM image revealed apati-
face showed a homogenous surface include fine granules te spheroid on treated dentin tubule opening (Fig. 2) that
with 5-micron width of Ca, Si (for tri-calcium silicate) this observation was the same with Xin Li’ observation
and Cl (for calcium chloride). C, Zr and N represent (intra-tubular remineralization contained tubular occlu-
zirconium oxide, carbonate and inorganic ingredients sion). In Xin Li study, resin -free cement had a higher
of Biodentine. After 7 days’ storage in HBSS, Zr and amount and rate of remineralization but this reminerali-
N peak disappeared, Si peak intensity was reduced and zation after 6 months did not reach 100 % even after six
Na, Mg and P peak found in solution. Material surface months. The differences between results may be due to
covered with globular deposits and P peak was raised. the differences in methodology.
Ca/P atomic ratio after 7 days reached 10.82 (24). In the Atmeh’s study on the remineralization potential of Bio-
present study, Ca, Si, Cl was seen in the primary com- dentine and glass ionomer (Fuji IX), on completely
ponent, but Zr, C and N were not seen. After 7 days, the demineralized dentin showed the mineral formation of
Si peak diminished and P peak was increased too but Na Ca-P like Raman peak related to apatite and most flo-
and Mg peak was not seen. The Ca/P atomic ratio of the rescence intensity was represented by Biodentine treated
present study after 7 days had reached 6.85 in Biodenti- dentin (26). Like the present study, Biodentine represen-
ne and 6.89 in Theracal. These differences could be due ted matrix remineralization, intra and peritubular mi-
to HBSS storage solution that has Ca in its composition. neralization in BSE-SEM. In addition, the apatite peak
In a Zhao et al. study, remineralizing effect of CPP-ACP found on normal dentin and dentin treated by Biodenti-
add to Glass ionomer was analyzed. It showed that the ne is completely similar (Fig. 3). Therefore, the forma-
remineralizing ability of CPP-ACP modified glass iono- tion of other Ca- P mineral components like amorphous
mer was improved (25). The present study also indicated calcium phosphate, octa calcium phosphate, di calcium
the same result. Dissolution of ACP in solution lead to a phosphate di hydrate or tri calcium phosphate was im-
super saturated solution, providing a nucleation site for plausible.
Ca, phosphate deposition and apatite formation. With Corral Nunez et al. assessed the bioactive properties of
the increase of pH, more formation of COO- group (pol- Biodentine (27). They concluded that incorporation of
yacrylate), form higher calcium polyacrylate. bioactive glass Nano-particles into Biodentine improve
Xin li et al. studied chemical interplay and reminerai- its bioactivity, after 7 days’ immersion in SBF. The me-
zation potential of resin -free hydraulic calcium silicate dian Ca/P molar ratio of Biodentine was 15.32 (Ca=35.4
cement (hCSCs) (Biodentine and ProRoot MTA) and and P =1.8 Wt%). Biodentine formed an apatite layer on
resin base calcium silicate cement (Theracal) at dentin its surface and inter-tubular precipitation of deposits. At
through Feg-EPMA and Raman Micro Spectroscopy present study, the median Ca/P molar ratio of Biodentine
(23). The result was the same as the present study. All was 15.83 (Ca=43.65 P=1.24 Wt%).
types of cements have remineralization effect after one Fathy’s study on remineralization ability of Biodentine
week but any different storage period was not signifi- and Theracal shown that Biodentine has a higher ability
cant. Unlike the present study, lower remineralization to remineralize the artificial carious dentin with signi-
was attained by resin base Theracal cement even after 6 ficantly higher mineral contents and both have signifi-

Fig. 2: SEM image from Biodentine treated dentin. Pointer shows apatite deposition on opening of
dentinal tubule and occlusion of it.

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Fig. 3: FTIR comparative analysis of intact dentin (A), demineralized dentin (B), CPP-ACP treated dentin (C),
Biodentine treated dentin (D), demineralized dentin 7 days in PBS (E), TheraCal treated dentin (F).

cantly induced remineralization on demineralized dentin FTIR analysis. According to SEM-EDX analysis, demi-
after 1 week or 6 months’ incubation period (28). Accor- neralization dentin can remineralize by bioactive cement
ding to the result of the present study, all three commer- (Biodentine and Theracal) and CPP-ACP (Bio-remine-
cial cement had the same result in terms of reminerali- ralization), but there are no significant differences be-
zing ability after 7 days (Ca=13.9, P=4.71 in Biodentine, tween the groups.
Ca=17.78, P=7.95 in Theracal and Ca=21.82, P=8.37 in
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Conflict of interest
The authors have declared that no conflict of interest exist.

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