Cent. Eur. J. Chem.

• 11(10) • 2013 • 1657-1667

DOI: 10.2478/s11532-013-0297-1

Central European Journal of Chemistry

Synthesis, characterization
and bioevaluation of partially stabilized
cements for medical applications
Research Article

Georgeta Voicu1, Alina I. Bădănoiu1*,

Ecaterina Andronescu1, Carmen M. Chifiruc2
1
Department of Science and Engineering
of Oxide Materials and Nanomaterials,
Faculty of Applied Chemistry and Material Science,
Politehnica University of Bucharest,
011061 Bucharest, Romania

2
Department of Microbiology and Immunology,
Faculty of Biology, University of Bucharest,
060101 Bucharest, Romania

Received 11 February 2013; Accepted 21 May 2013

Abstract: Materials for dental applications, i.e., white mineral trioxide aggregate (WMTA) and partial stabilized cements (PSC) were obtained
using the sol-gel method. The presence of ZnO or/and CaF2 additions in the starting mixture induced changes in the composition,
morphology and grindability of PSCs as compared with WMTA. The presence of foreign elements (Zn or F) in the crystalline lattice of
mineralogical phases, increased their grindability. Thermal analysis (TG&DTA) was used to assess the kinetics of hydration process in
binding systems based on WMTA/PSCs. The presence of foreign elements in PSCs systems increases the reactivity vs. water of these
materials and consequently, the compressive strength developed after 28 days of hardening at 37°C are higher as compared with WMTA.
The in vitro bioevaluation results (trypan blue staining, eukaryotic cells cycle assay by flowcytometry) accounted for a high biocompatibilty
of the obtained materials demonstrating their potential use for biomedical applications.
Keywords: Partial stabilized cement • Sol-gel synthesis • Hydration and hardening processes • Compressive strength • Cytotoxicity- eukaryotic
cell cycle stages
© Versita Sp. z o.o.

1. Introduction responsible for cement setting [9,10]. Although the

mineralogical composition of portland cement and MTA
Mineral trioxide aggregate (MTA) is a biocement is similar, the substitution of MTA with PC is presently
developed for surgical endodontic treatment, mainly for discouraged [4]. Two types of MTA were used for
retrograde filling and perforation repair [1-5]. The main endodontic treatments, i.e., of gray (GMTA) and white
requirements for a material with such applications are: (WMTA) colour [4,8,11]. The lighter colour of WMTA is
good sealling ability, quick setting, dimensional stability mainly due to a lower content in colouring ions, such as
and biocompatibility [4-7]. MTA consists of mineralogical iron, as well as its smaller particles size [4,11]. WMTA is
phases also present in portland clinker (PC), i.e., calcium preferred for dental restoration operations for aesthetical
silicates – Ca3SiO5 (C3S), Ca2SiO4 (C2S) and calcium purposes.
aluminate – Ca3Al2O6 (C3A) [3,4,8]. The development Some studies reported a long setting time and poor
of mechanical strength is determined in these systems handling properties of MTA [1,5,12], therefore a new
mainly by C3S and C2S hydration, while the C3A is material called Partial Stabilized Cement (PSC) has

* E-mail: [email protected]
1657
 Synthesis, characterization and bioevaluation
of partially stabilized cements for medical applications

been developed [5]. This material consists of a silicate The materials used for the sol-gel synthesis were:
cement in which transition elements (Co, Cr and Zn) are calcium nitrate (Ca(NO3)2•4H2O, >99.0%, Reag.
added in order to create crystal defects and high energy ACS., Sigma-Aldrrich, Germany), aluminium nitrate
state which favours the decrease of the cement setting (Al(NO3)3•9H2O, >98.5%, Reag. ACS, Sigma-Aldrrich,
time. Germany), tetraethyl-orthosilicate (C6H16O3Si, 99.0%,
The synthesis of C3S, C2S and C3A via solid-state Reag. ACS., Sigma-Aldrrich, Germany- TEOS), zinc
reactions consists in the sintering of stoichiometric nitrate (Zn(NO3)2•6H2O, >98%, Reag. ACS., Sigma-
mixtures of oxides or carbonates at high temperatures Aldrrich, Germany) and calcium fluoride (CaF2,
for long time [9,10]. Moreover, this type of synthesis >99.9%, Reag. ACS., Sigma-Aldrrich, Germany). These
implies multiple consecutive thermal treatments and materials were dosed in order to obtain a stoichiometric
intermediary material grinding; these operations mixture corresponding to the compositions presented in
could lead to introduction of material impurities and Table 1.
consequently to its colour and other properties change. The main steps of the sol-gel synthesis were the
Therefore the sol-gel method was used to synthesize following: calcium nitrate, followed by aluminium nitrate,
C2S [13,14], C3A [15], GMTA [16] and WMTA [17,18]. The zinc nitrate or/and calcium fluoride were dissolved in
main advantages of this synthesis route (as compared 180 mL water, under magnetic stirring, until a clear
with the conventional one) are lower thermal treatment solution was obtained. TEOS was hydrolysed using a
temperatures and higher purity of the resulted products molar ratio TEOS: water of 1:4. The two solutions were
[13-18]. mixed and continuously stirred at 60°C for 4 hours, and
Preliminary results showed the possibility to then kept the next 96 hours at 70°C, to facilitate the water
synthesize WMTA with good in vitro biocompatibility evaporation and to accelerate the polycondensation
by a sol gel route [18]. In order to obtain materials with reaction, resulting in the formation of a viscous gel. This
high reactivity vs. water and consequently with a faster gel was then dried at 120°C for 420 hours and the final
development of mechanical strength, two methods were product was a white powder [18].
studied: i) preparation of accelerated portland cement The powders were pressed in pellets and thermally
(APC) by WMTA mixing with CaCl2 addition [19] and ii) treated at 1350°C, for 30 minutes. Rapid cooling was
synthesis by sol-gel route of PSCs with ZnO, CaF2 or performed in air.
Zn+ CaF2 mixtures. The choice of these two additions in In order to obtain the cements these materials were
PSC synthesis was based on the fact that Zn, in small grinded for two hours in a laboratory planetary mill.
quantities is generally harmless to human body and is a
key element for bone development [5,17,20], while CaF2 2.2. Materials analysis
is a well known fluxing agent [21,22] and is a caries The white powders resulted by gel drying were
preventing agent [23,24]. analysed by thermal analysis (DTA/TG/DTG) and by
In this paper we present the influence of ZnO or/ X-ray diffraction analysis (XRD). Thermal analysis
and CaF2 additions on the composition, morphology was performed using a Shimadzu DTG-TA-60, in the
and grindability of PSCs synthesized by a sol gel route, 20-1000°C temperature range, with a heating rate
as well as on the hydration and hardening processes of 10°C min-1, in air. X-ray diffraction analysis was
and properties developed by these materials, including performed using a Shimadzu XRD 6000 diffractometer,
in vitro cytotoxicity and influence of the eukaryotic cell with Ni-filtered CuKa radiation (l=1.5406 Å), with scan
cycle. step of 0.02° and counting time of 0.6 s step-1.
The elemental composition of materials (WMTA
and PSCs) obtained from the thermal treatment was
2. Experimental procedure assessed by X ray fluorescence using a Rigaku ZSX
Primus XRF machine. The material’s compositions
2.1. Synthesis of WMTA and PSCs presented in Table 2 are similar to those calculated
One WMTA composition (K) and four PSC compositions based on the designed mineralogical composition
(KZ, KZF and KF) – Table 1, were synthesized using the (Table 1).
sol-gel route, previously described by Voicu et al. [18]. The mineralogical composition was assessed
The advantage of this method, as compared with the by XRD and their morphology by scanning electron
one proposed by Wang et al. [16] is the use of a lower microscopy (SEM) using a HITACHI S2600N equipment.
temperature thermal treatment (1350°C) of shorter The specimens for SEM analysis were covered with a
duration (30 minutes). thin silver layer deposited by dc-sputtering.

1658
G. Voicu et al.

Table 1. Designed mineralogical composition of WMTA and PSCs.

Material 3CaO·SiO2 (C3S) (%) 2CaO·SiO2 (C2S)(%) 3CaO·Al2O3 (C3A) (%) ZnO (%) CaF2 (%)

WMTA K 70 24 6 - -

KZ 70 24 6 1 -

PSCs KZF 70 24 6 1 0.5

KF 70 24 6 - 0.5

Table 2. Elemental compositions of synthesised biocements.

Composition K KZ KZF KF
(%wt.)
Clinker

Calculated
F 0 0 0.24 0.24

Al 1.20 1.19 1.19 1.20

Si 12.50 12.38 12.31 12.44

Ca 50.68 50.17 50.18 50.68

Zn 0 0.80 0.80 0

O 35.62 35.47 35.29 35.45

Assessed by XRF

F 0 0 0.24 0.24

Al 1.18 1.17 1.18 1.17

Si 12.47 12.37 12.27 12.42

Ca 50.66 50.15 50.17 50.66

Zn 0 0.79 0.80 0

O 35.69 35.52 35.34 35.51

The granulometric characteristics (i.e., Rosin- 2.3. Biological assays

Ramler-Sperling diagrams) of the powders resulted 2.3.1. Cells
after two hours of grinding in the laboratory planetary HCT8 line (ECAC90032006) and human diploid
mill were assessed by laser granulometry by means of a cells (HDC) were used in our experiments. HCT8
Malvern Mastersizer 2000 laser granulometer. line was cultivated in RPMI 1640 (Gibco, NY, SUA)
Free lime content of materials thermally treated at supplemented with 10% heat-inactivated bovine serum
1350°C for 30 minutes was assessed by EDTA method and penicillin/streptomycin at 37°C with 5% CO2. Diploid
as prescribed in European and corresponding national fibroblasts were cultivated in RPMI 1640 (Gibco, NY,
norm - SR EN 196-2:2006 [25]. SUA) supplemented with 10% heat-inactivated bovine
The hydration and hardening processes of WMTA/ serum and penicillin/streptomycin at 37°C with 5%
PSCs were assessed by XRD and thermal analysis of CO2.
pastes (solid/water ratio was 1 g per 0.33 mL), after
28 days of curing. The hydration was stopped by 2.3.2. Cytotoxicity Assay
acetone washing and drying at 60°C for 10 hours. Cytotoxicity was performed using Trypan blue staining
The compressive strength was assessed on paste after the treatment with 100 µg mL of the tested cements.
specimens - cylinders (f=10 mm and h=10 mm), cured In brief, a freshly prepared solution of 50 µL Tripan
the first day in a mould at 37°C and after demoulding blue (0.05%) in distilled water was mixed with 50 µL of
submerged, up to 28 days, in distilled water at 37°C. each of the cellular suspension during 5 min, spread

1659
 Synthesis, characterization and bioevaluation
of partially stabilized cements for medical applications

DTA uV 24 h at 37°C, 5%CO2 and in humid condition. Thereafter,

50
a cells were harvested, washed in phosphate saline buffer
(PBS, pH 7.5), fixed in 70% cold ethanol and maintained
0 K at -20oC, overnight. Each sample was washed in PBS,
treated with 100 µg mL-1 ribonuclease A (RNAse A) for
-50 15 minutes and coloured with 10 µg mL-1 propidium
62
iodide by 1 hour incubation at 37°C. After cell staining
KZF with propidium iodide, the acquisition was done using
-100 567
643 Epics Beckman Coulter flowcytometer. Data were
-150
analysed using WinMDI 2.9 software and expressed
as fractions of cells in the different cell cycle phases
552 611
[28,29].
-200
0 200 400 600 800 1000
Temp.(oC)

25
TG (%) 3. Results and discussion
b DTA, TG and DTG curves of the white powders resulting
20 from gel drying are presented in Figs.1 and 2.
On the DTG curve of WMTA (K), are present the
following processes, correlated with the information
15
provided by DTA curve: calcium and aluminium nitrates
dehydration and decomposition – effect with maximum
10 at 331°C, followed by calcium hydroxide decomposition
– effect with maximum at 497°C, anhydrous Ca(NO3)2
5 KZF melting (effect with maximum at 564°C) and finally
the effect from 849°C attributed to the decomposition
K of calcium carbonate, formed by partial carbonation of
0
calcium hydroxide [9,10,30,31].
0 200 400 600 800 1000
The DTA curves of KZ and KF (Fig. 2a) present
DTG (m g/s) K similar effects in the range of 400-700°C. A higher weight
0
loss recorded in this temperature range (see also TG –
849

Fig. 2b) for the specimen KZ suggests the presence of

-0,005
KZF higher amount of calcium nitrates.
564

The calcium nitrates are formed in these compositions

-0,01
due to the high concentration of NO3- in the starting
497

602

solution; consequently partial re-precipitation of calcium

-0,015
nitrates could occur. The presence of calcium nitrates
(Ca(NO3)2- JCPDS [07-0204] and Ca(NO3)2•2H2O-
497

-0,02
JCPDS [27-0087]) in the dry gels is confirmed also
331

-0,025
by the XRD spectra- Fig. 3. CaCO3 assessed on DTA
curves cannot be identified with certainty on the XRD
386

c
-0,03
spectra, most probably due to its low quantity (under
0 200 400 600 800 1000 XRD detection limit).
Tem p.(oC)
The free lime values assessed on WMTA and PSCs
Figure 1. DTA (a), TG (b) and DTG (c) curves of dry gel (white
obtained by thermal treatment at 1350°C for 30 minutes
powders) of K and KZF.
are presented in Table 3. Accepted free lime values are
onto a microscope slide and covered with a coverslip. not greater than 2% as it adversely affects the quality of
Nonviable cells became blue-stained. At least 200 cells cement [9]; as it can be seen from Table 3, all values are
were counted per treatment [26,27]. well below this limit.
The XRD spectra of WMTA and PSCs are presented
2.3.3. Cell cycle in Fig. 4. The main mineralogical compounds formed by
For cell cycle analysis, HDC or HCT8 cell lines were thermal treatment are: calcium silicates (3CaO•SiO2 -
treated with 100 µg cement mL-1, and maintained for JCPDS [42-0551], JCPDS [76-0623], JCPDS [31-0301]

1660
G. Voicu et al.

Table 3. Characteristics and properties of synthesised biocements.

Specimen K KZ KZF KF

Free lime* - (%) 1.05 0.32 0.28 0.41

Average C3S crystallite size*- d (nm) 24.54 35.38 38.51 28.91

Compressive strength** - Cs (MPa) 8.57±0.42 13.39±0.46 14.44±0.56 13.76±0.57

* assessed on materials thermally treated at 1350oC for 30 minutes; ** assessed on cement pastes hardened for 28 days at 37oC

DTA uV TG (%)
50 25
a b
0
KF 20

-50 62
655

KZ 15
KZ
560

647

-100
582

10
-150
KF
574

5
-200
590

-250 0
0 200 400 600 800 1000 0 200 400 600 800 1000
Tem p (oC) Temp.(oC)

Figure 2. DTA (a) and TG (b) curves of dry gels (white powders) of KZ and KF.

the C3S crystal lattice [22]. The incorporation of foreign

ions determines also the increase of the crystallite size.
Table 3 presents the average size of C3S crystallites
calculated with Debye-Scherrer equation [32] i.e.,
evaluated from the line broadening of 29.5(29.6)°,
30.7(30.8)° and 51.7(51.8)° peaks.
The morphology of WMTA and PSCs was assessed
using scanning electron microscopy (SEM) – Fig. 6.
As it can be seen on Figs. 6a and 6b in SEM
micrographs of K specimen, polyhedral C3S crystals
(average size 6-8 mm) and round C2S crystals
(average size 2-6 mm) were identified. In case of the
specimen with ZnO addition (KZ), the SEM micrographs
(Figs. 6c and 6d) showed agglomerations of small
crystals of 3-6 mm average size. For the specimens with
Figure 3. XRD spectra of dry gels (white powders). CaF2, the amount of liquid (molten) phase increased
(Figs. 6e-6h) as compared with K and KZ specimens.
and 2CaO•SiO2- JCPDS [33-0302], JCPDS [33-0303], These results are in good correlation with other literature
JCPDS [70-0388]) and calcium alumiante (3CaO•Al2O3- data, fluorides being well known as mineralizers and
JCPDS [38-1429], JCPDS [32-0150]).The XRD peak fluxing agents [22,30].
in the window 2q=51-53° (Figs. 4 and 5) can provide The influence of ZnO and/or CaF2 additions on the
information about 3CaO·SiO2 (C3S) polymorphism [9] grindability of obtained clinkers (WMTA and PSCs) can be
as well as the presence of foreign ions in its crystalline estimated considering the granulometric characteristics
structure. The shifting of the C3S peak at 2q= 51.86o of the powders formed after 30 minutes of grinding, in
(CuKa radiation) - recorded for specimen K, to lower similar conditions (Fig. 7). The particle sizes of PSCs
values for the PSCs (i.e., 51.72° - KZ, 51.76° - KZF (KZ, KZF and KF) are smaller than those of WMTA (K),
and 51.8° - KF) suggests the presence of foreign ions in i.e., the grindability of these materials is higher. This can

1661
 Synthesis, characterization and bioevaluation
of partially stabilized cements for medical applications

5000 A - 3CaO.SiO2;

A,B
B - 2CaO.SiO2;

A,B
4500 C- 3CaO.Al2O3;

A
A

A
4000

A
A,B

A
C

C
B
KF

A
A

A
C

B
3500

Intensity (counts)
3000

2500
KZF
2000

1500
KZ
1000

500
K
0
14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58

2 q (Degrees)
Figure 4. The XRD spectra of materials obtained by thermal treatment at 1350°C for 30 minutes.

900
K -1350
800 KZ -1350
KZF - 1350
700 KF - 1350
Intensity (counts)

600

500

400

300

200

100
51 51,25 51,5 51,75 52 52,25 52,5 52,75 53

2 q (Degrees)

Figure 5. C3S peaks in the XRD window 2ϴ=51-53°.

be explained by the incorporation of foreign elements in of crystalline anhydrous phases (C3S, C2S and C3A),
the silicates and aluminate structures, thus affecting the as well as of portlandite (calcium hydroxide), the main
microsymmetry and electrostatic relations, the chemical crystalline product formed by calcium silicates hydration.
bonds or/and coordination of ions in the structure [33]. The small amount of calcium carbonate present in the
The increase of grindability is important in connection pastes hydrated for 28 days is formed by portlandite
with both material reactivity vs. water as well as its carbonation, its sensitivity to CO2 being well known
aesthetical properties – the degree of whiteness of the [9,30].
material increases with the increase of its fineness. The thermal analysis can provide quantitative
The hydration and hardening processes in WMTA information regarding both crystalline hydrates
and PSCs systems were assessed by XRD and DTA/ (portlandite and calcium aluminates hydrates) as well
TG analysis on pastes hydrated for 28 days. The as poorly crystalline hydrates such as calcium silicates
XRD spectra presented in Fig. 8 show the presence hydrates (C-S-H) [9,10,30]. The DTA and TG curves

1662
G. Voicu et al.

(a) (b)

(c) (d)

(e) (f)

(g) (h)
Figure 6. SEM micrographs of: K - a and b; KZ – c and d; KZF - e and f; KF – g and h.

1663
 Synthesis, characterization and bioevaluation
of partially stabilized cements for medical applications

d
c
b
a

Figure 7. Rosin-Rammler-Sperling diagram of materials grinded for 30 minutes in similar conditions: a- K; b-KZF, c-KZ; d-KF.

5000 A - 3CaO.SiO2; B - 2CaO.SiO2;

CH

CH- Ca(OH)2; CC-CaCO3

A,B, CH
4500

4000

A, CH
A, B,CC

CH
CH

CH
A, B

3500
B

A
Intensity (counts)

KF 28 d.
3000

2500
KZF 28 d.
2000

1500

K 28 d.
1000

500
KZ 28 d.
0
15 20 25 30 35 40 45 50 55 60

2 q (Degrees)
Figure 8. XRD spectra of pastes hydrated 28 days.

DTA uV TG (%)
60
a 29
40 b
20
K 28 d. 27
0
25
-20

-40 685
142 23
KZ 28 d.
70 856
-60 657
21
-80 KZ 28 d.
-100 19 K 28 d.
-120
487 17
-140
455
-160 15
0 200 400 600 800 1000 0 200 400 600 800 1000
Temp.(oC) Temp.(oC)

Figure 9. DTA (a) and TG (b) curves of K and KZ pastes hydrated 28 days.

1664
G. Voicu et al.

TG (%)

29
b
27

25

23

21
KZF 28d.

19

17 KF 28 d.

15
0 200 400 600 800 1000
Temp.(oC)

Figure 10. DTA (a) and TG (b) curves of KZF and KF pastes hydrated 28 days.

100
(a) 100
(b)
90 90
80 80
70 DHC-Control 70 HCT8- Control
60 60
K,3d K,3d
% 50 % 50
KZ,3d KZ,3d
40 40
KZF,3d KZF,3d
30 30
KF,3d KF,3d
20 20
10 10
0 0
Go/G1 S G2/M Go/G1 S G2/M

Figure 11. Influence of WMTA (K) and PSC (KZ, KZF and KF) pastes hydrated for 3 days on the cellular cycle phases of: a – DHC; b- HCT 8 cells.

recorded on pastes hydrated for 28 days are presented of hydration processes as compared with WMTA (K
in Figs. 9 and 10. Three endo-effects could be seen on specimen). The higher reactivity vs. water of PSCs, as
DTA curve: compared with WMTA, is the direct consequence of the
- the large effect from 70-82°C and the shoulder from modification of crystalline structure, morphology and
142-164°C are most probably due to the superposition grindability induced by the ZnO and CaF2 additions.
of the following processes: loss of humidity and water The compressive strength of PSCs specimens
bound in calcium silicates hydrates (C-S-H) and calcium are higher than those recorded for WMTA (see
aluminate hydrates; Table 3), which is in good agreement with the information
- the endo-effect from 455-487°C is due to the obtained by DTA&TG.
portlandite dehydration, and The dye exclusion test was used to determine the
- the large endo-effect from 657-685°C (and the number of viable cells present in a cell suspension after
shoulder at 856-852°C) is attributed to the decomposition 24 hours treatment with the obtained cements (pastes
of CaCO3, with different crystallinity degrees, formed by hydrated 3 and 28 days). This test is based on the
Ca(OH)2 carbonation during specimens preparation and principle that live cells possess intact cell membranes
curing. that exclude certain dyes, such as trypan blue, eosin,
The results presented in Table 4 for the PSCs (KZ, or propidium iodide, whereas dead cells do not, so that
KZF and KF), show that the weight loss values recorded these substances accumulate inside the cells [31].
between 20-1000oC are higher when compared The cell cytotoxicity was calculated from the tripan
with WMTA (K). Also, calculated portlandite content blue assay on two cellular cell lines, i.e., HCT8 and
considering (or not) carbonation influence, is higher in DHC. The cytotoxicity induced by the tested compounds
PSCs specimens, suggesting an increase of the kinetics on the two cellular lines was reduced, ranging from 7%

1665
 Synthesis, characterization and bioevaluation
of partially stabilized cements for medical applications

Table 4.Weight losses recorded on TG curves and calculated Ca(OH)2 content for pastes hydrated for 28 days.

Specimen Weight loss in temperature (oC) range: Ca(OH)2 content* (%) Corrected Ca(OH)2 content** (%)
20-1000 400-575 575-1000

K 20.27 5.60 5.61 23.02 32.45

KZ 22.51 6.21 5.98 25.53 35.58

KZF 23.48 6.29 7.04 25.86 37.70

KF 22.32 6.45 6.52 26.52 37.48

* calculated based on the weight loss recorded between 400-575°C; ** calculated considering also Ca(OH)2 converted in CaCO3 (based on weight loss
between 575-1000°C)

100
(a) 100
(b)
90 90
80 80
70 DHC-control 70 HCT8-control
60 60
K,28d K,28d
% 50 % 50
KZ,28d KZ,28d
40 40
KZF,28d KZF,28d
30 30
KF,28d KF,28d
20 20
10 10
0 0
Go/G1 S G2/M Go/G1 S G2/M

Figure 12. Influence of WMTA (K) and PSC (KZ, KZF and KF) pastes hydrated for 28 days on the cellular cycle phases of: a – DHC; b- HCT 8 cells.

in case of HCT8 to 15% in case of DHC line. However, composition, morphology and grindability of PSCs as
no significant difference was observed between the 3 compared with WMTA.
and 28 days hydrated pastes. For PSCs, the increase of C3S crystallite size and
The eukaryotic cell cycle consists of two basic the shifting of its XRD peaks (as compared with WMTA),
stages: interphase and mitosis (M). The interphase has suggests the presence of foreign elements (Zn or/and
three stages called Gap 1 (G1), Synthesis (S), and Gap F) in the C3S crystal lattice with positive influence on
2 (G2). In the G1 phase, the cell is growing, replicating the materials grindability; the increase of grindability is
cytoplasmic organelles and preparing for replication of important in connection with both material reactivity vs.
its DNA, which is synthesized during S phase. During water as well as its aesthetical properties. The degree of
the G2 phase cells are preparing for division, which whiteness increases with the increase of fineness.
occurs in the M phase of the eukaryotic cell cycle. The Thermal analysis (TG&DTA) showed an increase
influence of different substances on the eukaryotic cell of the hydration rate in the binding systems with PSCs
faith might be cell cycle phase-dependent, which is a (as compared with WMTA) with direct consequences
reason why this test is used for the in vitro assessment on the mechanical strength of synthesised materials.
of the cytotoxicity. Compressive strength increases of 55-68% were
With respect to the influence of the tested substances recorded after 28 days of hardening at 37°C for the
on the cellular cycle of the two lines, no significant studied PSCs as compared with WMTA.
changes were observed in the two tested cellular lines The in vitro bioassays results demonstrate a high
on the pastes hydrated for 3 days (Fig. 11) and for biocompatibilty of the obtained materials demonstrating
28 days (Fig. 12). their potential for biomedical applications.

4. Conclusions Acknowledgements
The results presented in this paper show that white This paper is supported by the Sectoral Operational
mineral trioxide aggregate (WMTA) and partial Programme Human Resources Development,
stabilized cements (PSC) can be synthesized by sol-gel financed from the European Social Fund and by the
method. The presence of ZnO or/and CaF2 additions Romanian Government under the contract number
in the starting mixture determines the modification of POSDRU/89/1.5/S/64109.

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G. Voicu et al.

References

[1] M. Torabinejad, T.F. Watson, T.R. Pitt Ford, [19] G. Voicu, A.I. Bădănoiu, E. Andronescu, C. Bleotu,
J. Endod. 19, 591 (1993) Rev. Chim. 63, 1031 (2012)
[2] S.J. Lee, M. Monsef, M. Torabinejad, J. Endod. 19, [20] W-H. Wang, F-H. Lin, Y-L. Lee, C-P. Lin , J. Biomed.
541 (1993) Mater. Res. 81A, 195 (2007)
[3] J. Camilleri et al., Dent. Mater. 21, 297 (2005) [21] I. Teoreanu, T. van Huynh, S. Stoleriu, Rev. Roum.
[4] H.W. Roberts, J.M. Toth, D.W. Berzis, D.G. Charlton, Chim. 48, 947 (2003)
Dent. Mater. 24, 149 (2008) [22] L. Kamici, A. Seimenon-Masseron, A. Ghomari,
[5] F.H. Lin, W.H. Wang, C.P. Lin, Biomaterials 24, 219 Z. Derriche, C.R. Chimie. 9, 154 (2006)
(2003) [23] H.U.V. Gerth, T. Dammaschke, E. Schafer,
[6] T.R.P. Ford, D.J. McKendrey, Oral Surg. Oral Med. H. Zuchner, Dent. Mater. 23, 1521 (2007)
Oral Pathol. 79, 756 (1995) [24] H.T. Dean, Publ. Health. Rep. 53, 1443 (1938)
[7] S.K. Vasudev, B.R. Goel, S. Tyagi, Endodont. 15, [25] Romanian Standard, SR EN 196-2:2006, Methods
12 (2003) of testing cement - Part 2: Chemical analysis of
[8] S.J. Song, F.K. Mante, W.J. Romanov, S. Kim, cement (2006) (in Romanian)
OOOOE(Oral Surg. Oral Med. Oral Pathol. Oral [26] A.M. Grumezescu, A.M. Holban, E. Andronescu,
Radiol. Endod.) 102, 809 (2006) A. Ficai, C. Bleotu, M.C. Chifiriuc, Lett. Appl.
[9] H.F.W. Taylor, Cement Chemistry (Academic Press, NanoBioScience. 1, 77 (2012)
London, 1997) [27] C. Saviuc, A.M. Grumezescu, C. Bleotu, A. Holban,
[10] M. Georgescu, A. Bădănoiu, Cement Concrete C. Chifiriuc, P. Balaure, V. Lazar, Biointerface Res.
Comp.19, 295 (1997) in App. Chem. 1, 111 (2011)
[11] S. Asgary, M. Parirokh, M.J. Eghbal, S. Stone, [28] A.M. Grumezescu, E. Andronescu, A. Ficai,
F. Brink, J. Mater. Sci Mater. Med. 17, 187 (2006) C. Bleotu, M.C. Chifiriuc, Biointerface Res. in App.
[12] T. Dammaschke, H.U.V. Gerth, H. Züchner, Chem. 2, 438 (2012)
E. Schäfer, Dent. Mater. 21, 731 (2005) [29] O. Banu, C. Bleotu, M.C. Chifiriuc, B. Savu,
[13] D. Stephan, P. Wihelm, Z. Anorg. Allg. Chem. 630, G. Stanciu, C. Antal, M. Alexandrescu, V. Lazǎr,
1477 (2004) Biointerface Res. in App. Chem. 1, 72 (2011)
[14] G. Voicu, C.D. Ghiţulică, E. Dinu, E. Andronescu, [30] A. Bădănoiu, J. Paceagiu, G. Voicu, J. Therm. Anal.
Rev. Rom. Mater. 41, 229 (2011) Calorim. 103, 879 (2011)
[15] G. Voicu, C.D. Ghiţulică, E. Andronescu, Mater. [31] A. Migdal-Mikuli, J. Hetmanczyk, L. Hetmanczyk,
Characterization, 73, 89 (2012) J. Therm. Anal. Calorim. 89, 499 (2007)
[16] W-H. Wang, Y-L. Lee, C-P. Lin, F.-H. Lin, J. Biomed. [32] H.P. Klug, L.E. Alexander, (Eds.), X-Ray Diffraction
Mater. Res. 85A, 964(2008) procedures for polycrystalline and amorphous
[17] F. Ndong, S. Sadhasivam, F-H. Lin, S. Savitha, materials (John Wiley and Sons, New York, 1974)
W-H. Wang, C-P. Lin, International Endod. J. 45, 687
557 (2012) [33] L. Opoczy, V. Gavel, Int. J. Miner. Process. 74,
[18] G. Voicu, A.I Bădănoiu, C.D. Ghiţulică, S129 (2004)
E. Andronescu, Digest J. Nanomat. Biostruct. 7, [34] W Strober, Current Protocols in Immunology,
1639 (2012) A.3B.1–A.3B.2 (2001)

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