Professional Documents
Culture Documents
Biodentine
Publications and
Communications
2005 - 2010
Dec. 2010
TITLE YEAR AUTHORS REFERENCE
Impedance methodology: A new way to characterize the
setting reaction of dental cements
2010
Cyril Villat, V.X. Tran, Nelly Pradelle-Plasse,d, Pierre
Ponthiaux, Francois Wenger, Brigitte Grosgogeat,
Pierre Colon
Dent Mater 26 (2010) : 1127-1132
Ca
SiO
, CaCO
, ZrO
.
With time (after 3 months), the sealing ability of G-bond
Interface (SEM)
Cohesive fracture (B), cement (A), G bond
III (C).
C
A
B
30/74
Time Micro hardness
2H 51.5 ( 1.75)
1D 63.14 ( 1.94)
7D 72.19 ( 6.38)
30D 69.46 ( 1.45)
Table II: Average of hardness (HVN) and standard deviation (into brackets)
Interfaces Dye penetration (%) at D0 Dye penetration (%) at D90
Enamel / BIODENTINE
TM
17.65 (4.35) 19.86 (10.72)
Dentin / BIODENTINE
TM
10.46 (3.23) 14.84 (5)
Table III: Tooth BIODENTINE
TM
INTERFACES
% of microleakage
Adhesive systems Dye penetration (%) at D0 Dye penetration (%) at D90
Xeno
rd
RD94
X100
X100
48/74
A clinical study of a new Ca3SiO5-based material indicated as
a dentine substitute
2009
G.Koubi, J.C. Franquin, P. Colon.
Abstract in Clin. Oral Invest 2009 + poster Conseuro 2009
(Seville, Spain March 12-14th 2009)
49/74
50/74
A Clinical Study of a New Ca A Clinical Study of a New Ca
3 3
SiO SiO
5 5
- -based Material based Material
Indicated as a Dentine Substitute Indicated as a Dentine Substitute
G.F. KOUBI
1
, J.C. FRANQUIN
1
, P. COLON
2
1
Dpt. of Operative Dentistry and Endodontics, University of Marseilles, France.
2
Dpt. of Operative Dentistry and Endodontics, University of Paris 7, France.
OP065
Aim Aim
A new Ca
3
SiO
5
-based material (Biodentine RD94, Septodont) has been developed as a dentine substitute before direct and indirect posterior fillings. A recent study
conclude that this new material is in vitro biocompatible. It could be used as a pulp capping agent and as a bulk restorative material at the same time. A three-year follow-
up randomized multicentric clinical study was initiated to evaluate: 1) its longevity and in vivo biocompatibility as temporary restoration vs a resin composite (Z100, 3M),
and 2) its ability to be combined with an adhesive filling.
Materials and Methods Materials and Methods
Longevity and Biocompatibility in vivo
Three hundred and thirty-four (n = 334) patients (162 female and 172 male), aging from 18 to 80 years, with a
mean age of 47 years, were selected to participate in this study.
Distribution of material (Biodentine RD94 versus Z100) was randomized by means of a computer-generated
randomization list. One dentist, well experienced with both materials, placed all restorations.
Vital first and second permanent premolars and molars with Class I or Class II lesions were included. For
restorations with Z100 (3M, St Paul, MN, USA) inserted according to the manufacturers recommendation , no
cavity liners for indirect pulp capping, and no cavity bases were applied in addition to the adhesive (All Bond
2, Bisco, Ill., USA).
Composition of Biodentine RD94 (Septodont, France)
5900 osc./min
Purified water,
calcium chlor
LIQUID
200 mL
Mixed 25 s.
Linea Tac
Tricalcium silicate,
calcium carbonate,
calcium oxide
POWDER
1 g
For BiodentineRD 94, the new Ca
3
SiO
5
-based cement (Septodont, Saint-Maur, France), developed as a bulk dentine substitute before direct and indirect posterior fillings,
no special chemical procedures were applied on the mineralized walls.
Ability to be combine with an adhesive filling
When necessary, a definitive restoration was applied with Z100, in the same conditions previously described. The reason of the new definitive treatment was noted (wear,
fractures, loss of contact point), as well as the longevity of the temporary restoration.
Evaluation
The restorations were evaluated direct after placement (baseline), 15 days, and 6, 12, 24, 36 months. Each restoration was evaluated with slightly modified USPHS criteria
for the following characteristics: anatomical form, marginal adaptation, colour matching, marginal staining, surface texture, and secondary caries. The cavity form, the
depth and the clinical dimension of the cavities were also noticed. Radiographs were taken for assessments of proximal integrity and presence of recurrent caries. Intra-
oral colour slides were taken at baseline and at each recall period with a Nikon 70, equipped with medical lens Nikon 105 mm.
Results Results
The clinical study is still on-going. These results of the distribution of restorations by patients age are summarized in Figure 1, Figure 2, Figure 3.
18-29 y: 21.55%
30-39 y: 20.65%
40-49 y: 22.18%
50-59 y: 16.77%
60-69 y: 15.56%
70-80 Y: 3.29%
Figure 1. Distribution of restorations by patients age
Total RD 94 Z 100
0
100
200
300
Class I
Class II
Class I
Class II
Figure 2. Distribution of restorations by cavity class
0
100
200
300
400
Total RD 94 Z 100
PM
M
PM
M
Figure 3. Distribution of restorations by group of teeth
Longevity and Biocompatibility
On the 334 patients able to be recalled at 6 months, 254 came back for this control, given a recall percentage of 76 % and 48 % for d+1 year.
At the baseline, 24 teeth with deep cavities have to support a pulp capping with Biodentine RD94. At this time, 20 teeth have been checked, all of them with a positive pulp vitality, after
healing periods from 15 days to 26 months
Before 6 months, no Biodentine RD94 need to be replaced, and all restorations evaluated in this study at d+15 and d+6 months demonstrated acceptable clinical performance within the
evaluation period based on the Alfa and Bravo ratings for clinical satisfactory restorations.
After 6 months, some clinical failures were noted with Biodentine
TM
RD94 because of a too rapid occlusal wear.
48 restorations with Biodentine
TM
RD94 have to be replaced by a definitive restoration in Z100.
Ability to be combine with Adhesive filling Z100
None of these 48 retreated patients has lodged negative complaint for pain, unpleasant physiologic or pathologic sensations. The calcium silicate cement was easily preserved on all
dentinal walls, and the operator could easily prepare well calibrated cavities, without any infiltration or secondary caries. All the retreated teeth present a positive vitality test without any
hypersensitivity, and all of them have kept Biodentine
TM
RD94 on the dentinal walls, as bulk or liner in a sufficient quantity to permit directly a normal cavity preparation. No gap and no
secondary decay were observable between the temporary product Biodentine
TM
RD94 and the dentinal walls, and the preparation of the cavity for the definitive restoration was easy.
In these 48 cases, with complementary Z100 restorations, 28 have been yet controlled and considered satisfactory, after a mean period of 14 months. The second series of definitive
restorations in Z100 inserted on temporary filling will be evaluated with the same methods previously described, and then compared to the first series of Z100.
Conclusion Conclusion
Three years after the beginning of the study, the results seems to indicate that the new Ca
3
SiO
5
-based material called Biodentine
TM
RD94 could be used as dentin
substitute for definitive dentinal treatment, in restoration of posterior teeth. It could be used as a pulp capping agent and as a bulk restorative material at the same
time. The mean duration of Biodentine
TM
RD94 restorations was 12 months, with a minimum of 6 months and maxima of 35 months. As marginal leakage and
secondary decays remain a concern in adhesive dentistry, Biodentine
TM
RD94 can be preserved if re-intervention is required, and seems compatible with an
adhesive filling. This new product seems to include the most important qualities for a dentine substitute : biocompatibility and longevity. It could find rapidly a
place in the therapeutic practitioners equipment.
SEVILLE, SPAIN, MARCH
12th 14th 2009
253
137
116
33
81
48
RD94 Z100
233
124 109
101
45 56
RD94 Z100
Biodentineas bulk Biodentineas liner
Bibliography Bibliography
P. Laurent, J. Camps, M. De Mo, J. Dejou, I. About. Induction of specific cell responses to a Ca3SiO5-based posterior restorative material. Dental Materials 2008;24:1486-1494.
C. Boinon, Collage sur un substitut dentinaire base de silicate de calcium, Thse de Doctorat en Chirurgie Dentaire, Marseille, Juin 2006.
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Induction of specific cell responses to a Ca3SiO5-based
posterior restorative material
2008
Laurent P, Camps J, De Mo M, Djou J, About I. Marseille,
France.
Dent Mater. 2008 Nov;24(11):1486-94.
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dental materi als 2 4 ( 2 0 0 8 ) 14861494
avai l abl e at www. sci encedi r ect . com
j our nal homepage: www. i nt l . el sevi er heal t h. com/ j our nal s/ dema
Induction of specic cell responses to a Ca
3
SiO
5
-based
posterior restorative material
Patrick Laurent
a
, Jean Camps
a
, Michel De M eo
b
, Jacques D ejou
a
, Imad About
a,
a
Laboratoire IMEB - ERT 30, Facult e dOdontologie, Universit e de la M editerran ee, 27 Boulevard Jean Moulin,
13355 Marseille Cedex 05, France
b
Laboratoire de Biog enotoxicologie et Mutagen` ese Environnementale (EA 1784), Facult e de Pharmacie,
Universit e de la M editerran ee, Marseille, France
a r t i c l e i n f o
Article history:
Received 23 January 2007
Received in revised form
16 December 2007
Accepted 25 February 2008
Keywords:
Ca
3
SiO
5
-based dental cement
Biocompatibility
Genotoxicity
a b s t r a c t
Objectives. A Ca
3
SiO
5
-based cement has been developed to circumvent the shortcomings
of traditional lling materials. The purpose of this work was to evaluate its genotoxicity,
cytotoxicity and effects on the target cells specic functions.
Methods. Ames test was applied on four Salmonella typhimuriumstrains. The micronuclei test
was studied on human lymphocytes. The cytotoxicity (MTT test), the Comet assay and the
effects on the specic functions by immunohistochemistry were performed on human pulp
broblasts.
Results. Ames test did not show any evidence of mutagenicity. The incidence of lympho-
cytes with micronuclei and the percentage of tail DNA in the Comet assay were similar to
the negative control. The percentage of cell mortality with the new cement as performed
with the MTT test was similar to that of biocompatible materials such as mineral trioxide
aggregate (MTA) and was less than that obtained with Dycal. The new material does not
affect the target cells specic functions such as mineralization, as well as expression of
collagen I, dentin sialoprotein and Nestin.
Signicance. The new cement is biocompatible and does not affect the specic functions of
target cells. It can be used safely in the clinic as a single bulk restorative material without
any conditioning treatment. It can be used as a potential alternative to traditionally used
posterior restorative materials.
2008 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.
1. Introduction
Commonly used direct restorative materials for Class I and II
cavities are resin composites and amalgams [1,2]. In the early
1980s, amalgamrestorations were reported to release mercury
vapors which may be harmful to the environment, the dentist
as well as the patient [3].
Direct composite restorations have gradually been used
to replace amalgam for anterior restorations and small- to
C and pro-
cessed for immunohistochemistry. The effect of the materials
on the cytodifferentiation was evaluated by studying the spe-
cic protein expression of control cells compared to that of
cells cultured with the medium after being in contact with
the test material [20].
2.7.1. Immunohistochemistry
The cells were permeabilizedfor 15 minwith0.5%TritonX-100
in PBS. Primary antibodies were diluted in PBS containing 0.1%
Bovine Serum Albumin (BSA). The incubation with primary
antibodies was performed overnight at 4
C. Anti-collagen I
antibodies were used at 40 g/ml and anti-nestin antibody
at 5 g/ml. Anti-dentin sialoprotein antibody was diluted
1:200 in PBS. Immunostaining was revealed using the labeled
streptavidin-biotin kit (LSAB; Dako Corporation, Carpinteria,
CA, USA) according to the manufacturers instructions. Glyc-
ergel was used as a mounting medium (Dako Corporation).
Controls were performed by omitting primary antibodies or
incubationwithunrelatedprimary antibodies (cytokeratin19).
All controls were negative.
2.8. Genotoxicity assays
2.8.1. Ames test
S. typhimurium TA97a, TA98, TA100, and TA102 strains were
grown overnight from frozen cultures in Oxoid nutrient broth
No. 2 for 1012 h. Mutagenicity assays were performed as
described [21]. The genotype of each S. typhimurium tester
strain was conrmed in each experiment, and negative and
positive controls were routinely included.
After the preparation and setting of the cement, it was
ground to prepare a stock solution prior to testing by adding
60 mg of the cement in 1 ml of Nutrient Broth No. 2 (NB 2)
mediumor DMSOsolvent for 24 hat 37
C
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dental materi als 2 4 ( 2 0 0 8 ) 14861494 1489
for 48 h and revertant colonies were counted with an auto-
mated colony counter (Spiral System Instruments, Bethesda,
MS, USA). The experiments were carried out in the presence
and in the absence of an S9 fraction isolated from liver of
phenobarbital/-naphtoavone-treated rats. This S9 fraction
(4%) was routinely included in an S9-Mix, and the amount of
protein was adjusted to 1.25 mg protein per plate. A substance
was qualied positive if it induced a dose-related and repro-
ducible increase inthe numbers of revertants or twice as many
spontaneous revertants per plate [22].
2.9. Micronucleus test
This work was performed on lymphocytes obtained by
vein puncture from 6 healthy non-smoking donors, after
informed consent, and collected in glass tubes containing
lithium heparin anticoagulant according to Digue et al. [23].
Briey, cultures were carried out by adding 0.7 ml of whole
blood to 9.3 ml of X-VIVO
TM
Medium (Bio-Whittaker, Bel-
gium) supplemented with 25% fetal calf serum (Gibco, Life
Technologies
TM
, Germany), heparin (50 U/ml), and antibiotics
(penicillin 100 Ul/ml and streptomycin 100g/ml). The cells
were stimulated with phytohemagglutinin (1 mg/ml), a spe-
cic mitogen agent of human T-lymphocytes. The cells were
then cultured for 72 h at 37
C in a humidied atmosphere
containing 5% CO
2
.
The Ca
3
SiO
5
cement extract was prepared as described
above inthe culture mediumor DMSOandaddedtothe culture
at 24 h. The cells were directly exposed to serial dilutions (1%,
2.3%, 3.7%, and 5%) of the cement extracts for 48 h. Negative
control was achieved by adding DMSO at a nal concentration
of 0.1%. MitomycinC, used as a reference genotoxic agent, was
used as positive control 5g/ml. Cytochalasin B was added to
the culture (5g/ml) 44 h after PHA stimulation.
The cultures were stopped at 72 h and the cells harvested
by centrifuging (10 min at 360g). They were then treated
by a mild hypotonic treatment (1 min in KCl 0.075 M) and
immediately xed with methanol:acetic acid (3:1). This x-
ing step was repeated twice after 20-min storage at 4
C. Cells
were smeared on pre-cleaned microscope slides and air-dried.
Staining was performed with 5% Giemsa in Milli-Q water for
15 min.
Stainedslides were codedandscoredby light microscopy at
400 magnication. For each slide, 1000 Giemsa-stained bin-
ucleated lymphocytes with a well-preserved cytoplasm were
scored for the presence of micronuclei. In the micronucleated
binucleated cells, the number of MN per cell was recorded.
Micronuclei were expressed in terms of micronucleated cells
per 1000 binucleated lymphocytes. All the slides were exam-
ined twice by the same scorer. As a measure for toxicity, the
binuclearity index (BI) was determined by scoring the binu-
cleated cells for 1000 lymphocytes (mono- and binucleated
cells) and linked to the percentage of lymphocytes that pro-
duced complete cell division for the different drugs tested,
and then provided an index of cytotoxicity [24]. An extract
of a material was considered positive if at least a three-fold
increase of the numbers of micronuclei over negative controls
was observed at one or more dilutions of the original extract
[25,26].
2.10. Single-cell gel (Comet) assay
The Ca
3
SiO
5
cement extract was prepared and put in MEM
medium (60 mg/ml) for 24 h at 37
C, layered
onto a pre-coated slide with 1.5%regular agarose, and covered
with a coverslip. After brief agarose solidication in a refrig-
erator, the coverslip was removed and the slides immersed
in lysis solution (2.5 mol/l NaCl, 100 mmol/l EDTA, 10 mmol/l
TrisHCl buffer pH 10, 1% sodium sarcosinate with 1% Triton
X-100, and 10% DMSO) for about 1 h. Prior to electrophore-
sis, the slides were left in alkaline buffer (pH >13) for 20 min
and electrophoresed for another 20 min, at 25 V (0.86 V/cm)
and 300 mA. After electrophoresis, the slides were neutral-
ized in 0.4 mol/l TrisHCI (pH 7.5) xed in absolute ethanol,
and stored at room temperature until analysis blindly in a
uorescence microscope at 400 magnications. In order to
minimize extraneous DNA damage from ambient ultraviolet
radiation, all steps were performed with reduced illumina-
tion. Anautomatic analysis system(Comet Assay II; Perceptive
Instruments, Haverhill, UK) was used to determine DNA dam-
age. Tail moment (product of tail DNA/total DNA by the center
of gravity) was considered to estimate DNA damage from 50
cells per treatment.
3. Results
3.1. Determination of the toxicity with or without
dentin disc interposition
When the toxicity was evaluated indirectly through a dentin
slice, the analysis of variance failed to showa statistical differ-
ence between the new cement, Pro Root MTA, and Dycal (ns)
(Table 1). None of the materials was cytotoxic. However, when
the toxicity was evaluated without dentin slice interposition,
the analysis of variance showed a statistically signicant dif-
ference among the three materials (P <0.001). The Duncan test
Table 1 Cytotoxicity after indirect contact between the
materials and culture medium through a dentin disc
New Ca
3
SiO
5
cement MTA Dycal
Undiluted 0 8% 0 9% 0 8%
50% 0 4% 0 4% 0 4%
10% 0 4% 0 3% 0 4%
The newCa
3
SiO
5
cement, MTA, and Dycal were applied on the coro-
nal side of the dentin slices in Plexiglass devices with pulp pressure
simulation. After 24 h, the culture media in contact with the pul-
pal side of the dentin slices were used to determine cell viability.
The pulp broblasts were incubated with these media (either undi-
luted, or diluted in the culture medium to 50% or to 10%) for 24 h
before applying the MTT test on human pulpal broblasts. Opti-
cal density values of untreated control cultures normalized to 100%
was in the range of 0.90.95. The results are expressed as mean cell
toxicityS.D.
57/74
1490 dental materi als 2 4 ( 2 0 0 8 ) 14861494
Table 2 Cytotoxicity after direct contact between the
material and culture medium
New Ca
3
SiO
5
cement MTA Dycal
Undiluted 0 8% 0 9% 22 10%
50% 0 5% 0 5% 10% 5%
10% 0 4% 0 3% 2 2%
The cytotoxicity of the new cement compared to MTA and Dycal on
human pulp broblasts was evaluated after 24 h contact between
the materials and the culture medium (either undiluted, or diluted
in the culture medium to 50% or to 10%) with the MTT test. Both
were less cytotoxic than Dycal (P <0.001). The results are expressed
as mean cell toxicityS.D.
showed that Dycal displayed a higher cytotoxicity than MTA
and the new Ca
3
SiO
5
cement (Table 2).
According to this study, a dilution of 10% was chosen for
studying the materials effects onbroblasts specic functions
because it has biological effects without being toxic.
3.2. Inuence of the two materials on pulp broblasts
differentiation into odontoblastic cells
Control cells expressed collagen I, dentin sialoprotein and
Nestin. Pulp broblasts secreted a mineralizd matrix and the
cells, particularly those contacting the mineralizd matrix,
expressed Nestin (Figs. 1 and 2).
Fig. 1 Effect of the new Ca
3
SiO
5
cement on pulp broblast
specic gene expression. Immunohistochemistry was used
to evaluate the effect of the new Ca
3
SiO
5
cement and MTA
on pulp cells specic genes expression. Control cultures
express collagen type I (a) and dentin sialoprotein (b).
When the media containing the new Ca
3
SiO
5
cement (c
and d) and MTA (e and f) extracts were added to the
cultures for 4 weeks, collagen I (c and e) and dentin
sialoprotein (d and f) were also expressed at a high level in
the pulp cells. Original magnications =10.
Fig. 2 Effect of the new Ca
3
SiO
5
cement on pulp cells
mineralization. Immunohistochemistry was used to
evaluate the effect of the new Ca
3
SiO
5
cement and MTA on
pulp cells differentiation and mineralization. Control
cultures express Nestin and secrete a mineralized matrix in
the form of nodules (a). When the media containing the
new cement (b) or MTA (c) extracts were added to the
cultures for 4 weeks, a mineralized matrix deposition was
also observed. Nestin was also expressed at a high level in
pulp cells and its expression was stronger in the mineral
nodules forming cells. Original magnications =10.
After adding the media containing extracts of the new
Ca
3
SiO
5
cement or MTA to the cultured pulp cells, collagen I,
dentin sialoprotein were strongly expressed by the pulp cells
(Fig. 1). Mineral nodule formation was also observed (Fig. 2).
Nestin was expressed by the cells but not in the mineral nod-
ules. The immunostaining intensity was always higher incells
forming the mineral nodules than the cells away from these
nodules.
3.3. Genotoxicity
Ames test did not show any evidence of mutagenicity of
the Nutrient Broth No 2 medium after being in contact with
the new cement, whatever the dilution of the test medium
(Table 3). The mutations observed with the new cement were
comparable to the spontaneous reverse mutations obtained in
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dental materi als 2 4 ( 2 0 0 8 ) 14861494 1491
Table 3 Mutation frequencies of Ames tester strains using the liquid preincubation assay
Metabolic activation
(S9 mix
a
)
Product Volume (l) Number of revertants/plate (meanS.D.)
TA 97a TA 98 TA 100 TA 102
+ NB No. 2 10 171 9 243 1364 382 17
+ DMSO 10 166 7 251 12513 355 16
NB No. 2 5 183 13 265 13511 402 18
DMSO 5 191 11 274 1389 423 26
+
New Ca
3
SiO
5
cement (NB No. 2 extract) 4 162 14 301 13514 360 10
6 177 5 261 1202 397 15
8 177 4 295 1325 351 7
10 192 4 274 15013 345 2
New Ca
3
SiO
5
cement (NB No. 2 extract) 2 215 11 251 16110 500 24
3 223 9 253 17221 424 36
4 225 15 251 16035 439 3
5 205 23 231 18212 517 44
+
New Ca
3
SiO
5
cement (DMSO extract) 4 170 19 292 1193 334 49
6 189 3 252 12613 376 3
8 175 2 288 1451 336 24
10 164 23 437 1365 314 11
New Ca
3
SiO
5
cement
(DMSO extract)
2 193 2 352 1493 421 5
3 186 5 375 1178 445 42
4 224 17 273 1406 463 26
5 173 8 301 1443 435 36
+ B[a]P 0.5 g 112137 42326 100087 679 28
ICR 191 0.02 g 55321 NT NT NT
2,4,7 TNFone 0.02 g NT 1653 NT NT
NaN3 0.5 g NT NT 58512 NT
MitC 0.2 g NT NT NT 365854
After preparation and setting of the cement, it was grinded prior to testing. 60 mg of the cement were placed in 1 ml of Nutrient Broth No 2
or DMSO solvent for 24 h at 37
C under mixing. The stock solutions from two independent experiments were tested in triplicate, and results
from both experiments in NB 2 and DMSO are presented. Increasing volumes of test samples (4, 6, 8 and 10l) were incubated with the each
of the bacterial strains for 60 min at 37
C under mixing. The mixture consisting of bacteria and a test compound was plated on plates in VB
medium at 37 C for 48 h and revertant colonies were counted. The experiments were carried out in the presence and in the absence of an S9
fraction. The test was qualied positive if it induced a dose-related and a reproducible increase of the numbers of revertants or twice higher
than the spontaneous revertants per plate. All data are expressed as means S.D. Positive controls were Benzo[a]pyrene (0.5g) with S9 MIX for
all strains. Positive controls were 2-methoxy-6-chloro-9-(3-(2-chloro-ethyl)aminopropylamino)acridine (ICR 191, 0.1g) for TA97a; 2,4,7-trinitro-
9-uorenone (2,4,7-TNFone, 0.02g) for TA98; sodium azide (NaN
3
, 1 g) for TA100 and mitomycin C (MitC, 0.05 g) for TA102 without S9 MIX.
NT: non-tested.
a
The S9 MIX included 4% S9, 4.2 mM NADP and 5.2 mM G6P.
the controls performed with the NB 2 and DMSO solvent. The
results show that the new Ca
3
SiO
5
cement does not induce
reverse mutations either withor without the S9 metabolic acti-
vation system. Similar results were obtained with all bacterial
strains tested.
Table 4 Micronucleated human lymphocytes count in
Ca
3
SiO
5
cement-treated cultures
Ca
3
SiO
5
cement dilution Micronucleated
lymphocytes (%) S.D.
1% 4.0 1.1
2.3% 4.0 1.1
3.7% 4.0 1.2
5% 4.2 1.2
Negative control
a
3.7 1.2
Positive control
b
16.0 6.0
***
Comparison with the control:
***
P <0.001.
a
Culture medium X-VIVO 10.
b
Mitomycin C 5g/ml.
The micronuclei test revealed that after incubating the
lymphocytes with different dilutions of the new cement, the
rate of lymphocytes with micronuclei was similar to that
obtained with the negative control. It ranged from 3.9% to
4.1% with increasing concentrations (15%) in aqueous or
hydrophobic medium. The positive control showed a rate of
16% (Table 4).
The Comet assay performedwithserial dilutions of the new
Ca
3
SiO
5
cement on human pulp broblasts revealed that the
percentage of DNA in the tail ranged from 12.59 for the 0.1%
dilution to 15.58 with undiluted medium. This percentage was
13.19 with the negative control and 46.52 with the positive
control (Table 5).
4. Discussion
The biocompatibility of the newcement is shown in this study
by the absence of cytotoxicity and genotoxicity and the fact
that the new material does not affect the cytodifferentiation
of human pulp broblasts in odontoblastic cells.
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1492 dental materi als 2 4 ( 2 0 0 8 ) 14861494
Table 5 Comet assay on human pulp broblasts
Ca
3
SiO
5
cement dilution Tail DNA (%) meanS.D.
0.1% 12.59 0.96
1% 13.31 0.88
10% 14.90 1.06
Undiluted 15.58 1.08
Negative control
a
13.19 0.96
Positive control
b
46.52 1.45
***
Comparison with the control: ***P <0.001. NS: non-signicant.
a
0.1% DMSO.
b
H
2
O
2
(13.2 mM).
Although Portland cements are known as non-toxic, in this
work, 3 tests were performed to evaluate the genotoxicity of
the new Ca
3
SiO
5
cement after solubilisation in hydrophilic or
hydrophobic conditions. These tests were performed because
the cement developed here contains a modied polycar-
boxylate in the superplastisizer. It has been reported that
polycarboxylate (Aqualox
Exp material MTA
72/74
Physical, chemical and mechanical behavior of a new material
for direct posterior fillings.
2005
J. DEJOU, J COLOMBANI and I. ABOUT. Marseille, France
abstract : European Cells and Materials Vol. 10. Suppl. 4,
2005 (page 22)
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European Cells and Materials Vol. 10. Suppl. 4, 2005 (page 22) ISSN 1473-2262
Physical, Chemical and Mechanical Behavior of a New Material for Direct
Posterior Fillings.
J. Djou, A Raskin, J Colombani & I. About
Laboratoire IMEB-ERT 30, UFR dOdontologie, Universit de la Mditerrane, Marseille,
FRANCE
INTRODUCTION: A new ceramic material
for direct restorative posterior fillings, a
Ca
3
SiO
5
-based Portland cement, has been
developed to circumvent the shortcomings of
the traditional filling materials. The purpose of
this work was to evaluate the marginal sealing
efficiency, the acid erosion and the effects of
aging in artificial saliva on its structure,
composition and compressive strength.
METHODS: The marginal sealing was
evaluated by the silver nitrate penetration
method without any surface treatment, with or
without aging in Fusayama artificial saliva.
The acidic erosion was evaluated daily in lactic
acid (0.02M) and sodium lactate (0.1M)
aqueous solution (pH 2.74) by measuring the
height loss, for a week.
Aging was evaluated in Meyer-modified
Fusayama artificial saliva1 (pH 5.3).
The height modification of the material was
evaluated for a week. Scanning electron
microscopy was used to examine and
characterize the surface of the sample before
and after aging. The possible dissolution of the
new material in the artificial saliva was
evaluated by measuring the concentration of
Si, Ca, Zr, and inorganic carbonate in the
artificial saliva after 1, 2, 3 and 4 weeks. The
compressive strength was measured 24 hours
after setting and after aging for seven and 28
days.
RESULTS: No difference in marginal sealing
was revealed between the new biomaterial and
the Z250-Optibond solo plus adhesive
restorative system. The same results were
obtained after aging for one week in artificial
saliva. The acid erosion increased with time.
This increase was less rapid than that obtained
with glass ionomer cement reported by
Nomoto R2,3. In artificial saliva there was no
erosion but deposition of white material on the
surface of the material. Scanning electron
microscopic analysis of this material revealed
needle-like crystals with an apatitic appearance
(figure.1).
Fig. 1: Needle-like crystals on the surface of
the material after aging in artificial saliva
The composition of this deposit determined by
X-diffraction analysis seems to confirm the
apatitic composition (ratio Ca/P = 1.6). This
correlates well with the analysis of the
elements in the solution, which reveals a
decrease of Ca concentration with time. There
was a slight but not significant release of Si.
The compressive strength was 136 (20.10) at
24 hours, increased to 169.74 (16.92) after 7
days and then was stable until day 28.
DISCUSSION & CONCLUSIONS: The
marginal sealing without any surface treatment
or adhesive system was equivalent to that of
the reference material used. In spite of the
acidic pH of the artificial saliva, the new
material showed no erosion and an increase in
the compressive strength. The deposition of
apatitic structures might increase the marginal
sealing of the material.
REFERENCES:
1
Reclaru L, Meyer JM.
(1994). Study of corrosion between a titanium
implant and dental alloys. J Dent; 22:159-68.
2
Nomoto R, McCabe JF. (2001). A simple acid
erosion test for dental water-based cements.
Dent Mater ;17(1):53-9.
3
Nomoto R, Uchida K, Momoi Y, McCabe JF.
(2003). Erosion of water-based cements
evaluated by volumetric and gravimetric
methods. Dent Mater.;19(3):240-4.
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