Basic Research—Technology
Solubility and pH Value of 3 Different Root Canal
Sealers: A Long-term Investigation
Kent Urban, Dr, med, dent,* Julian Neuhaus, cand, med, dent,*
David Donnermeyer, Dr, med, dent,* Edgar Sch€afer, Prof, Dr, med, dent,†
and Till Dammaschke, Prof, Dr, med, dent*
Abstract
Introduction: The aim of this study was to compare the
solubility and the change in pH of a tricalcium silicate–
containing sealer (BioRoot RCS [BR]), a mineral trioxide
aggregate–containing sealer (MTA Fillapex [MTA]), and
an epoxy resin–based sealer (AH Plus [AH]) during a
longer period of time. Methods: The solubility test
was carried out in accordance with ISO6876:2012. Three
hundred eighty sealer samples (n = 10 per group) were
stored for 6 months in double distilled water (AD) or
phosphate-buffered saline (PBS). The solubility was
determined on the basis of the mass difference in
percent. The pH measurement was carried out on the
basis of the experimental setup of the solubility test
with the aid of an electrode pH meter. The results
were evaluated for significance by using analysis of vari-
ance, Student-Newman-Keuls, and t test (P < .05).
Results: The highest solubility was found for MTA in
AD (4.65%  1.17%), followed by BR in AD
(3.88%  0.42%) (P < .05). Also in PBS, MTA
(3.24%  0.7%) showed a higher solubility than BR
(1.78%  0.5%) (P < .05). AH was virtually insoluble
during the entire period (0.5%  0.5%) (P < .05). The
pH decreased continuously over time for all samples. Af-
ter 6 months, BR showed the highest pH in AD (pH 10.3),
followed by MTA in AD (pH 8.8), BR in PBS (pH 7.5), AH
in PBS (pH 7.3), MTA in PBS (pH 7.2), and AH in AD (pH
6.3). Conclusions: The solubility of MTA was signifi-
cantly increased compared with BR and AH. Storage in
PBS significantly reduced solubility of MTA and BR. BR
showed a higher pH in both AD and PBS than MTA in
AD and PBS. AH did not cause any significant pH
change. (J Endod 2018;-:1–5)
Key Words
AH Plus, BioRoot RCS, long-term solubility, MTA Filla-
pex, pH value
From the *Department of Periodontology and Operative Dentistry, Westphalian Wilhelms-University, M€unster, Germany; and †Central Interdisciplinary Ambulance in
the School of Dentistry, M€unster, Germany
Address requests for reprints to Prof Till Dammaschke, Westphalian Wilhelms-University, Department of Operative Dentistry, Albert-Schweitzer-Campus 1, Building
W 30, 48149 M€unster, Germany. E-mail address: tillda@uni-muenster.de
0099-2399/$ - see front matter
Copyright ª 2018 American Association of Endodontists.
https://doi.org/10.1016/j.joen.2018.07.026
JOE — Volume -, Number -, - 2018
C alcium silicate cements
such as mineral trioxide
aggregate (MTA) or Bio-
Significance
Solubility of the tricalcium silicate–containing
sealer BioRoot RCS was in accordance with ISO
dentine (Septodont, St.
6876:2012, even after 6 months of immersion in
Maur-des-Fosses, France)
simulated body fluid. The pH of BioRoot RCS in
were introduced to the
PBS was in an alkaline range for about 4 months.
dental market for endodon-
tic treatments, eg, for the
repair of lateral root canal or furcal perforations. In addition, these cements can be
used for retrograde closure after surgical endodontics (apicoectomy) resection or for
apexification. Calcium silicate cements were reported to be biocompatible and bioactive,
eg, they promote bone regeneration (1). Because of these beneficial properties, calcium
silicate cements have also been recommended as root canal filling material (2). However,
as a root canal filling material these cements has some disadvantages; the procedure is tech-
nically demanding and difficult in narrow, curved root canals, and retreatment and a post
insertion are hardly possible. Therefore, in recent years, root canal sealers based on cal-
cium silicate have been developed, which are used together with gutta-percha for root canal
obturation.
The aim of a root canal filling is the three-dimensional bacteria and fluid tight seal
of the entire root canal system to prevent passage of microorganisms from coronal to
apical or vice versa (3, 4). Therefore, root canal filling materials should be more or less
insoluble to prevent dissolving by body fluids in the root canal. Besides the apical
foramen, numerous microscopic and macroscopic communications exist between
the root canal system and the periodontal ligament and the surrounding bone,
namely dentinal tubules, accessory foramina, and lateral canals (5). Thus, tissue fluid
can easily penetrate the root canal system, resulting in degradation of the sealer mate-
rial. Consequently, a low solubility of sealers in distilled water as stated in the ISO stan-
dard 6876:2012 is required (6). The long-lasting bacteria-proof seal of the root canal
depends mostly on the integrity of the sealer, not the core material (7, 8).
Various publications dealing with solubility of calcium silicate–based sealers can
be found in the current literature. Apparently a disadvantage of these sealers is that the
solubility is higher than that of conventional epoxy-based sealers (9–15). However, it is
still unclear how the solubility develops during a longer period of time. Long-term low
solubility is certainly important for the clinical success of root canal obturation.
It has been shown that the solubility of calcium silicate–based sealers can be ex-
plained by the release of OH and Ca2+ ions (9, 16, 17). Therefore, the pH of the
solutions was also determined.
Solubility and pH value of 3 Root Canal Sealers 1
Basic Research—Technology
The aim of this study was to measure long-term solubility and pH
value of a comparably new tricalcium silicate–containing sealer (Bio-
Root RCS; Septodont, St. Maur-des-Fosses, France) and a MTA-
containing sealer (MTA Fillapex; Angelus, Londrina, Brazil) in compar-
ison with a conventional epoxy resin–based root canal sealer (AH Plus;
Dentsply DeTrey, Konstanz, Germany) during a period of 6 months. The
null hypothesis was that BioRoot RCS and MTA Fillapex will show a
significantly higher solubility and pH value than AH Plus.
Materials and Methods
BioRoot RCS (LOT powder B 17884, liquid B 16766 B), MTA Fil-
lapex (LOT 40367), and AH Plus (LOT compound A 1607000406, com-
pound B 1607000533) were used. All sealers were mixed according to
the manufacturers’ instructions.
On the basis of data of a previous study (13), power calculation
using G*Power 3.1 (Heinrich Heine University, D€usseldorf, Germany)
indicated that the sample size for each group should be at least 9.
Thus, 10 samples were used for each group.
Solubility Test
The solubility was determined on the basis of the results obtained
after immersion of the samples in double distilled water (AD) (Phar-
macy of the University Hospital, M€unster, Germany) and in
phosphate-buffered saline solution (PBS) (Pharmacy of the University
Hospital, M€unster, Germany). The specimens’ change in weight was re-
corded. For all sample preparation, stainless steel ring molds with a
height of 1.6 mm (0.1 mm) and an internal diameter of 20.0 mm
(0.1 mm) were used. All molds were cleaned in an ultrasound
bath with acetone for 15 minutes. Thereafter, a copper wire was fixed
at each mold to hang the specimens in a glass dish in such way that
the surfaces did not touch and the materials remained undisturbed
in the dish. Before use all molds were weighed 3 times
(accuracy  0.0001 g; Sartorius 1801MPS, G€ottingen, Germany),
and the mean was calculated.
The ring molds were placed on a glass plate and filled to slight
excess with the mixed sealer material, avoiding air entrapment. To
ensure complete setting of all sealers before testing, samples were
immersed in physiological solution (PBS) at 37 C for 48 hours (17).
The proper setting was evaluated in a pretest.
After setting of the sealers, excess material was trimmed to the sur-
face level of the mold by using silicon carbide paper (600 grit). From
each sealer, 50 samples were prepared for immersion in AD and 50
samples for immersion in PBS (150 mL). In each case the 50 samples
were divided into 5 groups of 10 for immersion in AD or in PBS for
14 days, 1 month, 2, 4, and 6 months. Furthermore, 10 ring molds
for each group were prepared for immersion in AD or PBS without
any sealer and were used as negative control during the entire period
of the experiment. All samples were stored in an incubator (Heraeus,
Hanau, Germany) at 37 C and 95% relative humidity. A first fluid
TABLE 1. Solubility of BioRoot RCS, MTA Fillapex, and AH Plus in AD
14 d 1 mo
Material Mean (SD) Mean (SD)
BioRoot RCS 1.600b (0.193) 1.999b (0.318)
MTA Fillapex 3.382c (0.385) 3.397c (0.377)
AH Plus 0.131a (0.125) 0.230a (0.370)
Data expressed as mean percentages with standard deviation (SD) of weight loss for each material and for each immersion period. Values with different superscript letters were statistically different at P = .05
(analysis of variance and Student-Newman-Keuls test).
2 Urban et al.
change was performed on all samples after 24 hours. Subsequently,
all fluids were changed once a week.
The materials in their ring molds were weighed 3 times before the
immersion of the samples. The average reading was recorded. After im-
mersion the samples were weighed again 3 times, and the mass of the
cements was determined to the nearest 0.0001 g. The difference be-
tween the original weight of material and its final weight was recorded
to the nearest 0.0001 g. This difference in mass was calculated as a per-
centage of the original weight of the material, recorded to the nearest
0.001%. The solubility test is a modification of the methodology set
in ISO 6876:2012 (6), eg, solubility testing in PBS. The detailed exper-
imental setup has been described previously (7, 13, 18).
After the solubility test, the sealer samples were dried and sput-
tered with gold for scanning electron microscopy (SEM) and energy-
dispersive x-ray analysis (EDX). For x-ray photoelectron spectroscopy
(XPS) and inductively coupled plasma optical emission spectroscopy
(ICP-OES), appropriate samples were pulverized (19).
pH Value
The pH measurement was carried out in parallel and based on the
experimental setup of the solubility test. The pH value was determined
with an electrode pH meter (accuracy  0.01; Sartorius Basic Meter
PB-11, G€ottingen, Germany). Before each pH measurement, the accu-
racy of the pH meter was controlled with calibration solutions (pH 4, 7,
and 10; Merck, Darmstadt, Germany). In addition, after each individual
measurement, the electrode was cleaned with AD to avoid contamina-
tion of the subsequent fluid. The measurement was carried out at
34 C fluid temperature for all samples to simulate the situation in the
oral cavity. The first pH measurement was carried out after 12 hours,
then every 2 days, and after 14 days weekly before renewal of the test
liquids.
According to the Kolmogorov-Smirnov test, all data were distrib-
uted normally; therefore, they were analyzed by using analysis of vari-
ance and post hoc Student-Newman-Keuls and t test (P < .05).
Results
Solubility
BioRoot RCS and MTA Fillapex were soluble during the entire
investigation period. The highest solubility was found in MTA Fillapex
in AD (4.65%  1.17%), followed by BioRoot RCS in AD
(3.88%  0.42%). Overall, MTA Fillapex was significantly more soluble
than BioRoot RCS, whereas BioRoot RCS was significantly more soluble
than AH Plus at all times investigated and in AD as well as in PBS
(P < .05). AH Plus was virtually insoluble during the entire period
(0.5%  0.5%) (P < .05) (Table 1).
BioRoot RCS and MTA Fillapex were highly significantly more sol-
uble in AD than in PBS (P < .001), except MTA Fillapex after 6 months
(P < .05). For AH Plus no significant difference was found (P > .05)
(Table 2).
2 mo 4 mo 6 mo
Mean (SD) Mean (SD) Mean (SD)
2.537b (0.181) 2.970b (0.135) 3.882b (0.415)
4.054c (0.799) 4.143c (0.429) 4.649c (1.165)
0.141a (0.084) 0.202a (0.220) 0.541a (0.560)
JOE — Volume -, Number -, - 2018
 Basic Research—Technology
TABLE 2. Solubility of BioRoot RCS, MTA Fillapex, and AH Plus in PBS
14 d 1 mo 2 mo 4 mo 6 mo
Material Mean (SD) Mean (SD) Mean (SD) Mean (SD) Mean (SD)
BioRoot RCS 0.936b (0.183) 0.991b (0.174) 1.325b (0.193) 1.518b (0.395) 1.784b (0.493)
MTA Fillapex 1.844c (0.508) 1.717c (0.519) 1.922c (0.508) 2.199c (0.214) 3.242c (0.697)
AH Plus 0.110a (0.081) 0.192a (0.050) 0.232a (0.040) 0.211a (0.081) 0.240a (0.051)
Data expressed as mean percentages with standard deviation (SD) of weight loss for each material and for each immersion period. Values with different superscript letters were statistically different at P = .05
(analysis of variance and Student-Newman-Keuls test).

The control samples of empty molds did not change weight after
immersion in water or PBS, respectively, after 6 months.
On the surface of all BioRoot RCS (after 14 days) and MTA Fil-
lapex (after 1 month) specimens immersed in PBS, a white precip-
itate was visible. This precipitation was much more pronounced in
the BioRoot RCS group (Fig. 1). However, this precipitation could
not be determined in the AH group. The deposits adhered to the sur-
faces even after drying and were hard to remove. The main compo-
nents of the precipitate were analyzed by XPS and EDX, and the
minor constituents were identified with ICP-OES. Moreover, the
chemical composition of the precipitate was investigated by
XPS and the surface morphology by SEM (Fig. 1), as described

Figure 1. SEM micrograph of the sealer surface after 6-month storage in PBS or AD. Original magnification, 1000. The bar represents 10 mm. (A) BioRoot RCS
after 6 months in PBS; (B) BioRoot RCS after 6 months in AD; (C) MTA Fillapex after 6 months in PBS; (D) MTA Fillapex after 6 months in AD; (E) AH Plus after
6 months in PBS; (F) AH Plus after 6 months in AD.

JOE — Volume -, Number -, - 2018 Solubility and pH value of 3 Root Canal Sealers 3
Basic Research—Technology
previously (19). The precipitation was calcium hydroxyapatite in the same range as that reported by Pr€ullage et al. (13) with
(Ca10(PO4)6(OH)2)  n H2O). 1.763% and 1.785%, respectively. Thus, the results were reproducible.
The discrepancy between the findings of different studies concerning
pH Value solubility might be attributed to variations in methods used, eg, to dry
The pH decreased continuously over time for all samples. The pH the samples after having subjected them to solubility testing (15).
of BioRoot RCS in AD dropped from 12.1 to 10.3 after 6 months and A similar picture emerges for MTA Fillapex. Here too the values are
from 11.4 to 7.5 in PBS. In AD the pH for MTA Fillapex was 10.3 after controversial. Whereas some authors found a comparatively low solu-
12 hours and 8.8 after 6 months, whereas in PBS the pH dropped from bility of 0.452% to 1.76% (11, 13, 15) after 24 hours, others reported a
9.3 to 7.2. For AH Plus in AD the pH was 9.3 after 12 hours and 6.3 after high solubility of 11.1% (14) to 14.24% (12). Comparable higher
6 months, and in PBS it was 7.6 and 7.3, respectively (Table 3). values of about 15% were also reported after 7 days (9, 10). The
After 6 months, BioRoot RCS showed the highest pH (pH 10.3) in variety in MTA Fillapex solubility may be explained by the fact that
AD, followed by MTA Fillapex in AD (pH 8.8), BioRoot RCS in PBS (pH MTA Fillapex hardly sets completely under different conditions. This
7.5), AH Plus in PBS (pH 7.3), MTA Fillapex in PBS (pH 7.2), and AH sealer was unable to set within 1 week in a dry environment (17) or
Plus in AD (pH 6.3) (Table 3). did not set completely after 1 week even when stored at 37 C and
95% relative humidity (13), whereas a setting time of more than 19
Discussion hours was measured for samples immersed in Hank’s balanced salt so-
lution (HBSS) (17). It may be speculated that in studies where a high
Methods solubility was reported, the setting of MTA Fillapex may be incomplete.
In a previous study it was shown that BioRoot RCS was significantly In the present study a proper setting was evaluated in a pretest.
less soluble in simulated body fluid (PBS) than in AD. In addition, after In contrast to BioRoot RCS and MTA Fillapex, AH Plus was more or
14 and 28 days, a hydroxyapatite precipitate was observed on the sur- less insoluble, which is in accordance with the recent literature. In pre-
face of the sealer. Presumably because of this precipitation, the solubility vious studies the solubility of AH Plus was determined to range between
of BioRoot RCS decreased. Nevertheless, the solubility was significantly 0.045% and 0.8% (7, 9–11, 13–15, 18).
higher than that of a conventional resin-based sealer (AH Plus) (13). BioRoot RCS does not include a calcium phosphate phase; howev-
The increased solubility of calcium silicate–based sealers in water er, this phase was identified when the material was immersed in a phys-
has also been described by other authors (9, 10, 12, 14, 15). iological solution such as HBSS (17) or PBS (13). This deposit seems to
Therefore, the purpose of this study was to investigate how solubility impede a further increase of solubility. The SEM micrographs showed
develops during a long-term period of 6 months. It has been speculated significantly more precipitated crystals on the surface of BioRoot RCS
that the hydroxyapatite precipitate leads to a kind of self-sealing effect of than on MTA Fillapex (Fig. 1). Hence, it may be speculated that if a cal-
BioRoot RCS. Whether the precipitation is stable in the long term, or that cium silicate–containing sealer is immersed in simulated body fluid, the
it may lead to a further decrease in solubility, has been completely un- calcium ions combined with phosphate are promoting the formation of
clear so far. To the best of our knowledge, there is no study investigating a superficial layer of hydroxyapatite, which is able to fill open voids orig-
the solubility of sealers during such a long period of time. inated by the high solubility (14). If voids reduced over time in the pres-
The solubility tests performed in the present study followed to a ence of simulated body fluid, calcium silicate–based sealers in
great extent the methodology of ISO 6876:2012 (6). According to combination with gutta-percha may improve sealing ability by the depo-
ISO 6876:2012, the solubility test is required solely in AD (6). The sol- sition of hydroxyapatite (calcium phosphate) at the wall-dentin-sealer
ubility of calcium silicate–based sealers in AD does not predict the real interface (21). Such an apatite deposition on the sealer surface may
stability and integrity of such sealers in vivo. To provide a better under- compensate the solubility (14). Hence, it may be assumed that calcium
standing of what will happen in contact of the sealers to body fluid, a silicate–based sealer may have a self-sealing ability. Nevertheless, the
solubility testing in PBS was carried out here (13, 20). high solubility of calcium silicate–based sealers remains an important
issue requiring further research.
Solubility
Most studies on the solubility of the calcium silicate–based sealers
investigated in distilled water had a relatively short examination period, pH
making comparison with the values of this study difficult. The pH values of all sealers were higher when tested in AD than in
For BioRoot RCS the given solubility values are contradictory. PBS. Under a clinical point of view, it may be speculated that the pH
Although in a previous study the solubility of BioRoot RCS was values measured in PBS as simulated body fluid are more realistic
1.174% after 24 hours (13), other authors reported about 11.05% that those obtained in water. In AD, BioRoot RCS and MTA Fillapex
(15) and 14.2% (14) solubility after 1 day. The solubility found in showed an alkaline pH during the entire period of the experiments,
the present study after 14 days (1.600%) and 28 days (1.999%) are whereas AH Plus showed a slightly acidic pH after 4 months. In PBS,
BioRoot RCS showed an alkaline pH for 4 months, whereas MTA Filla-
pex had a neutral pH already after 2 months. AH Plus was more or less
TABLE 3. pH Values of BioRoot RCS, MTA Fillapex, and AH Plus in AD and PBS
pH neutral during the entire experimental time (Table 3).
BioRoot MTA The pH values given in literature for BioRoot RCS range from 11.7
RCS Fillapex AH plus Control after 3 hours (14) to about 11–12 after 1 day (14, 15, 22, 23) and 8.7
AD PBS AD PBS AD PBS AD PBS (14) to 12.7 (22), respectively, after 28 days. For MTA Fillapex, pH
12 h 12.1 11.4 10.3 9.3 9.3 7.6 7.5 7.5
values of 9.27–9.68 (12, 14, 24) after 3 hours can be found in the
14 d 11.9 10.8 10.1 8.8 8.9 7.6 7.5 7.5 literature. After 1 day the pH changed to 8.02 (15) and 9.5 (14),
1 mo 11.7 11.0 10.3 8.1 8.2 7.5 7.4 7.3 whereas after 7 days it was 7.76 (24) to 9.2 (14). After 1 month, values
2 mo 11.6 9.6 10.7 7.6 7.8 7.4 7.5 7.4 of 8.21 (12), 8.4 (14), and 11 (11) were reported.
4 mo 11.4 8.0 10.3 7.4 6.4 7.4 7.4 7.5 Overall, the pH values of the present study are in accordance with
6 mo 10.3 7.5 8.8 7.2 6.3 7.3 7.5 7.3
the data given in recent literature. It has to be kept in mind that to the

4 Urban et al. JOE — Volume -, Number -, - 2018


best of our knowledge, this is the first study evaluating the pH during a
period of 6 months and also in PBS. An alkaline environment during
such a long period of time as caused by BioRoot RCS may play a positive
role in apical healing. In particular, the alkaline pH of the sealer may
increase the pH value of cytoplasm and body fluid in the periapical re-
gion, which may be lowered because of inflammation. Furthermore, the
high pH may neutralize the lactic acid released from osteoclasts and
thus may contribute to the hard tissue formation by activating alkaline
phosphates (25), have antimicrobial effects inside the root canal (26),
prevent dissolutions of mineralized components of teeth and favor the
deposition of mineralized tissue (27–30), and activate the
solubilization of bioactive components from the organic matrix (31)
and the deposition of apatite (32, 33). Thus, it has been
demonstrated that BioRoot RCS stimulates the release of tissue
growth factors such as vascular endothelial growth factor, fibroblast
growth factor 2, and bone morphogenetic protein 2, which
participate in the mineralization process of bone (34). Even though
BioRoot RCS increased the pH value of cell culture medium to 11,
the sealer had a positive influence on the osteoblast cell metabolism
(bioactivity) and was biocompatible (23).
The null hypothesis of the present study was confirmed; BioRoot
RCS and MTA Fillapex showed a significantly higher solubility and pH
value than AH Plus during a period of 6 months.
Conclusion
The solubility of BioRoot RCS and MTA Fillapex increased during a
period of 6 months. However, this increase was significantly lower in
PBS than in AD. For BioRoot RCS in PBS the solubility was less than
3% after 6 months as required according to ISO 6876:2012 (6),
whereas MTA Fillapex was significantly more soluble. AH Plus was virtu-
ally insoluble during the entire period. Despite the higher solubility, the
pH of MTA Fillapex was lower compared with BioRoot RCS. The pH was
in an alkaline range for BioRoot RCS in PBS for about 4 months,
whereas for MTA Fillapex it was just for 1 month. AH Plus was not alka-
line.
Acknowledgments
The authors thank Priv.-Doz. Dr. Gunther Brunklaus, Institute
for Physical Chemistry, Westphalian Wilhelms-University, M€unster,
Germany for analyzing the white precipitation of BioRoot RCS and
MTA Fillapex after storage in PBS by XPS, EDX, ICP-OES, and SEM.
The authors deny any conflicts of interest related to this study.
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