Journal of Dentistry 43 (2015) 1448–1455

Contents lists available at ScienceDirect

Journal of Dentistry
journal homepage: www.intl.elsevierhealth.com/journals/jden

Monomer elution in relation to degree of conversion for different types

of composite
Pong Pongprueksaa,b , Jan De Muncka , Radu Corneliu Ducac, Katrien Poelsc ,
Adrian Covacid , Peter Hoetc, Lode Godderisc , Bart Van Meerbeeka ,
Kirsten L. Van Landuyta,*
a
KU Leuven BIOMAT, Department of Oral Health Sciences, KU Leuven (University of Leuven) & Dentistry, University Hospitals Leuven, Kapucijnenvoer 7, box
7001, 3000 Leuven, Belgium
b
Department of Operative Dentistry and Endodontics, Faculty of Dentistry, Mahidol University, No.6, Yothi road, Rajthawee, Bangkok 10400, Thailand
c
Centre for Environmental and Health, Department of Public Health and Primary Care, KU Leuven (University of Leuven), Kapucijnenvoer 35, 3000 Leuven,
Belgium
d
Toxicological Center, University of Antwerp, Universiteitsplein 1, D.S.551, 2610 Wilrijk, Belgium

A R T I C L E I N F O A B S T R A C T

Article history: Objectives: The aim of this study was to determine the degree of conversion (DC) and the monomer
Received 12 August 2015 release of three composite types when employed following a layer- and bulk-filling technique.
Received in revised form 14 October 2015 Methods: The release of monomers from a ‘conventional paste-like’ (Filtek Z250), a ‘conventional
Accepted 16 October 2015
flowable’ (Filtek Supreme XTE Flowable) and a ‘bulk-fill’ flowable composite (Filtek Bulk Fill Flowable)
from the same manufacturer (3M ESPE, Seefeld, Germany) was determined. Ten cylindrical specimens
Keywords: per composite were built, either in two 2-mm layers or in one 4-mm bulk. DC was measured at the
BisGMA
specimen top and bottom surface using micro-Raman spectroscopy, after which the specimens were
BPA
UDMA
immersed in 2 ml absolute ethanol for 24 h at 37  C. This solution was refreshed weekly during six weeks
TEGDMA and the concentration of BisGMA, BisEMA(6), BisPMA, UDMA, TEGDMA and BPA was determined by
BisPMA liquid chromatography/mass spectroscopy.
BisEMA(6) Results: DC at the specimen top and bottom was similar except for the bulk-fill technique, which resulted
Degree of conversion in significantly lower DC at the specimen bottom. The release of BisGMA and TEGDMA was initially very
Monomer elution high, but rapidly dropped in the second week. In contrast, the release of BisPMA and UDMA increased
LC/MS–MS initially, but then declined towards the sixth week. BisEMA(6) release was relatively steady over time. All
composites released small amounts of BPA. The total monomer release was significantly lower for the
layer- than the bulk-filling technique.
Conclusions: The slightly reduced degree of conversion at 4-mm depth resulted in a higher monomer
elution when the composite was applied following a bulk-fill application method.
Clinical significance: Applying a flowable and a bulk-fill composite following a bulk-fill application
method resulted in a significantly reduced degree of conversion at the bottom of polymerized composite
specimens when compared to a layer-application method. This reduced polymerization degree was
reflected in significantly increased monomer release.
ã 2015 Elsevier Ltd. All rights reserved.

1. Introduction more recently introduced ‘bulk-fill’ composites can be applied and

In order to polymerize sufficiently and to reduce polymeriza-
tion shrinkage stress, it has been recommended to apply dental
composite incrementally, commonly in 2-mm thick layers. The
* Corresponding author at: KU Leuven BIOMAT, Department of Oral Health
Sciences, KU Leuven (University of Leuven), Kapucijnenvoer 7, Blok A, Box 7001, B-
3000 Leuven, Belgium. Fax: +32 16 33 27 52.
E-mail address: kirsten.vanlanduyt@med.kuleuven.be (K.L. Van Landuyt).
cured properly in 4-mm thick layers. Filling cavities with
composite in bulk became feasible thanks to reduced shrinkage-
stress during polymerization and deeper curing. Typically, bulk-fill
composites contain new monomers [1], more translucent filler [2]
and new photo-initiator systems [3].
As bulk-fill composites are intended to fill deep cavities with the
cavity bottom close to the pulp, some biocompatibility concerns
may arise in particular when the composite would not cure
properly at the cavity bottom, so that monomers can leach out in

http://dx.doi.org/10.1016/j.jdent.2015.10.013
0300-5712/ ã 2015 Elsevier Ltd. All rights reserved.

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 P. Pongprueksa et al. / Journal of Dentistry 43 (2015) 1448–1455 1449

Table 1
List of the three composites investigated and their composition.

Composite Shade Compositiona,b Filler sizea Lot number

Filtek Z250 Universal A3 1–10 wt% BisGMA, <5 wt% TEGDMA, 1–10 wt% UDMA, 1– 0.01–3.5 mm zirconia/silica filler N446661
10 wt% BisEMA(6), 75–85 wt% silane treated ceramic filler (average 0.6 mm)
Filtek Supreme XTE Flowable A3 5–10 wt% BisGMA, 5–10 wt% TEGDMA, 15–25 wt% BisPMA, 0.02 mm non-agglomerated/ non- N445013
50–60 wt% silane treated ceramic filler, 5–10 wt% silane aggregated silica filler, 0.005–0.01 mm
treated silica filler zirconia filler
Filtek Bulk Fill Flowable A3 1–10 wt% BisGMA, <1 wt% TEGDMA, 10–20 wt% UDMA, 1– 0.01–3.5 mm zirconia/silica filler N426393
10 wt% BisEMA(6), 10–20 wt% BisPMA, 50–60 wt% silane (average 0.6 mm), 0.1–5.0 mm
treated ceramic filler, 5–10 wt% silane treated silica filler ytterbium trifluoride
a
According to technical information provided by the manufacturer 3M ESPE.
b
Abbreviations: BisGMA, bisphenol A-glycidyl methacrylate; TEGDMA, triethylene glycol dimethacrylate; UDMA, urethane dimethacrylate; BisEMA(6), bisphenol A
ethoxylateddimethacrylate; BisPMA, 2,2-Bis-(4-(3 methacryloxypropoxy) phenyl) propane.

the oral cavity and easily reach the pulpal tissues. Methacrylate
monomers are very reactive in nature, and in particular in-vitro
research has shown that they may adversely interact with oral
cells. It has been revealed that they may disturb the redox
homeostasis through the generation of reactive oxygen species,
thereby seriously disturbing vital cell functions [4]. They are not
only cytotoxic at high concentrations [5,6], but they have also been
associated with genotoxicity [7,8]. In this regard, the polymeriza-
tion efficiency is important, as composites with low polymeriza-
tion degree release more monomers [9].
Polymerization efficiency is best measured chemically in terms
of the degree of monomer conversion (DC) using Fourier Transform
infrared spectroscopy (FTIR) or micro-Raman spectroscopy
(mRaman). In particular, the ratio of DC measured at the bottom
to that at the top surface was found to correlate well with surface
micro-hardness, typically measured as an indirect evaluation of
the polymerization efficiency [10]. Curing depth is affected by
many factors such as, among others, the irradiated light-output,
the uniformity of the light bundle, the distance to the composite
surface, the light attenuation by the composite and the surround-
ing tissue, and the light transmittance through the composite [11–
13].
Apart from monomers, composites may also release other
compounds, the most controversial being Bisphenol A (BPA) [14–
17]. Since BPA is used to synthetize some very frequently used
monomers in composites, such as BisGMA and BisEMA, composites
may unintentionally contain BPA as a residue of the monomer
synthesis process. Being a mild xeno-oestrogen, BPA has mainly
been associated with reproductive toxicity [18].
The objective of this study was to determine DC and the release
of monomers of three different types of composite when employed
following a layer- and bulk-filling technique, and to determine
whether any relationship between DC and the release of
monomers may exist up to six weeks. The null hypotheses tested
were (1) that no difference in DC and in the overall release of
monomers was measured for the three different composite types
and (2) that the application technique employed (bulk filling
versus layer filling) did not influence the degree of conversion and
monomer release.
2. Materials and methods
A conventional restorative composite with a paste-like
consistency (Filtek Z250 Universal), a ‘conventional flowable’
composite (Filtek Supreme XTE Flowable) and a ‘bulk-fill flowable’
composite (Filtek Bulk Fill Flowable), all from the same manufac-
turer (3M ESPE, Seefeld, Germany), were selected (Table 1). The
monomer composition of the three composites is alike and could
be retrieved from the technical information provided by the
manufacturer (MSDS).
Cylindrical samples were prepared of each composite type
(n = 10) using a teflon mold (5-mm diameter, 4-mm depth) that
was filled with composite in two 2-mm thick layers or in one 4-mm
bulk. The top and the bottom were covered with a glass slide to
prevent oxygen inhibition. Each of the two layers and the bulk were
cured for 20 s using a polywave LED light-curing unit (Bluephase
20i, Vivadent Ivoclar, Schaan, Liechtenstein). Irradiance at the tip
was measured using the MARC1 Patient Simulator (BlueLight
Analytics, Halifax, NS, Canada) to be around 1.100 mW/cm2.
After polymerization, the specimen was immediately removed
from the mold and four Raman spectra were acquired from the
middle area of the top and bottom surface using micro-Raman
spectroscopy (mRaman; Senterra, Bruker, Ettlingen, Germany). The
surface was excited with a near-infrared (785 nm) laser of 50 mW
and analyzed through a 100 objective and 50-mm pin-hole
aperture. The collected spectra ranged from 50 to 3,500 cm 1 with
a resolution of 9–15 cm 1. The integration time of each spectrum
was set to 20 s with 2 co-additions. The CCD detector to obtain the

Table 2
Characteristics of the dental monomers and their derivatives investigated.

Molecule Name Molecular formula Molecular weight CAS- Supplier

number
BisGMA Bisphenol A-glycidyl methacrylate C29H36O8 512.6 1565-94-2 Sigma–Aldricha
BisEMA(6) Bisphenol A ethoxylateddimethacrylate C35H48O10 628.7 41637-38-1 ESSTECH Incb
BisPMA 2,2-Bis-(4-(3 methacryloxypropoxy) phenyl) propane C29H36O6 480.59 27689-12-9 3M ESPEc
UDMA Urethane dimethacrylate C23H38N2O8 470.56 72869-86-4 Sigma–Aldrich
TEGDMA Triethylene glycol dimethacrylate C14H22O6 286.32 109-16-0 Sigma–Aldrich
BPA Bisphenol A C15H16O2 228.29 80-05-7 Sigma–Aldrich
d16-BPAd Deuterated (d16) bisphenol A C15D16O2 244.38 96210-87-6 Sigma–Aldrich
d4-diethyl phthalated Deuterated (d4) diethyl phthalate C6D4C6H10O4 226.26 93952-12-6 Cambridge Isotope Laboratoriese
a
St Louis, MO, USA.
b
Fremont, CA, USA.
c
Seefeld, Germany.
d
Added to solvent as internal standard for LC/MS–MS analysis.
e
Andover, MA, USA.

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mRaman spectra possessed a 1024  256 pixel resolution, and was
cooled down thermo-electrically to a temperature of 65  C.
Results were processed using OPUS 7.0 software (Bruker, Ettlingen,
Germany). DC was calculated as the ratio of peak intensities of the
aliphatic 1639 cm 1 and aromatic 1609 cm 1 peaks in cured and
uncured materials.
After mRaman, the specimens were immediately immersed in
2 ml extraction solvent, containing absolute ethanol (99.99%; CAS:
64-17-5, VWR, Haasrode, Belgium) with d4-diethly phthalate and
d16-BPA added as internal standards (Table 2). The negative
control consisted of the extraction solvent. Care was taken to use
only glass containers and glass pipettes to avoid contamination.
Specimens were kept in an incubator at 37  C and the extraction
solvent was refreshed every week during six weeks. Stock
solutions of BisGMA, BisEMA(6), BisPMA, UDMA, TEGDMA and
BPA (Tables 2 and 3) were prepared by dissolving commercial
reference standards in the extraction solvent (Table 2), so as to
prepare calibration standards. Quantification of the different
compounds in the samples was performed by ultra-pressure
liquid chromatography (UPLC), in combination with tandem mass
spectrometry (MS–MS). UPLC/MS–MS analysis of the samples was
conducted on a Waters1 Acquity UPLCTM, coupled to a Waters1
Micromass Quattro PremierTM Mass Spectrometer (LC/MS–MS,
Acquity System, Waters Corporation, MA, USA) using electro spray
ionization (ESI). A 10 mL aliquot of each sample was introduced on
an Acquity UPLC BEH C18, 50 mm  2.1 mm, 1.7 mm column, held at
a temperature of 40  C.
For BisGMA, BisEMA(6), BisPMA, UDMA and TEGDMA, the
mobile phase used for the chromatographic separation was a
mixture of 0.1% formic acid in water (A) and 0.1% formic acid in
Table 3
Molecular structure of the monomers investigated.
Molecule Molecular structure
BisGMA
BisEMA(6)
BisPMA
UDMA
TEGDMA
BPA
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acetonitrile (B) using the following gradient: the program was
starting at 10% B, increasing linearly to 100% B for 2 min, then held
for 2 to 2.5 min at 100% B, and finally brought back to the initial
status for 2.5 to 4.0 min. A flow rate of 0.35 ml/min was applied. The
analyses were performed in the positive ESI mode; a multiple
reaction monitoring (MRM) method was used with argon as the
collision gas.
For BPA, the mobile phase used was a mixture of 0.5% ammonia
in water (A) and 0.5% ammonia in methanol (B) using the following
gradient: the program was starting at 5% B, increasing linearly to
95% B for 3 min, then held for 3 to 3.7 min at 95% B, and finally
brought back to the initial status for 3.7 to 5.0 min. A flow rate of
0.50 ml/min was applied. For BPA, the analyses were performed in
the negative ESI mode; a MRM method was used with argon as the
collision gas.
The peak area was determined for each monomer and plotted
versus the concentration using linear regression analysis for
BisGMA (r2 = 0.988), BisEMA(6) (r2 = 0.904), BisPMA (r2 = 0.956),
UDMA (r2 = 0.891), TEGDMA (r2 = 0.981) and BPA (r2 = 0.987). The
calibration standards were used to quantify the monomer
concentration in the sample solutions. The monitored ion
transitions, the applied cone voltage, the collision energy for each
transition and the limit of quantification (concentration giving a
signal-to-noise (S/N) ratio  10) are reported in Table 4.
2.1. Statistical analysis
A linear mixed-effects model was constructed (nlme package,
R3.1.0, R Foundation for Statistical Computing, Vienna, Austria) to
assess DC. In this model, three fixed effects, namely ‘COMPOSITE’,
 P. Pongprueksa et al. / Journal of Dentistry 43 (2015) 1448–1455 1451

Table 4
Characteristics of each molecule detected in the analysis.

Molecule RTa (min) Parent ion (m/z) Daughter ion (m/z) Electrospray Cone (V) Collision energy (V) LOQb (mg/ml)
BisGMA 2.16 535.3 535.3 + 50 15 0.0057
BisEMA(6) 2.50 646.5 291.0 + 20 20 0.0625
BisPMA 2.64 481.4 261.2 + 20 10 0.0053
UDMA 1.95 493.7 493.7 + 50 10 0.002
TEGDMA 1.71 309.6 309.6 + 35 10 0.0008
BPA 2.28 226.9 211.8 – 40 20 0.013
d16-BPA 2.22 241.0 222.9 – 40 20 –
d4-diethyl phtalate 1.75 227.6 153.2 + 15 18 –
a
Retention time.
b
Limit of quantification.

‘LAYERS’ (layer- versus bulk-filling) and ‘SURFACE’ (top versus
bottom), were included, as well as all first order interaction terms.
The specimen measured was considered as a random effect. To
assess the difference in DC at the top and bottom surface, specific
contrasts were calculated from this model (contrast package,
R3.1.0, R Foundation for Statistical Computing). All tests were
performed at a significance level of a = 0.05. The absolute
monomer elution data were analyzed by one-way ANOVA and
Tukey’s multiple comparison test (a = 0.05). The BPA curve fitting
and any correlation between DC at the top/bottom surfaces and
total monomer release were calculated (R 3.1.0, R Foundation for
Statistical Computing).
3. Results
DC at the specimen top and bottom surface is presented in
boxplots for the three different composites investigated in the five
experimental groups (Fig. 1). For all composites, the mean DC
varied between 60 and 70%. DC measured at the top and bottom
surface were significantly different for all experimental groups (all
p values were smaller than 0.0013). The difference in top-bottom
DC was, however, mostly very small (<2%), in particular when the
composite was applied in layers. Despite being small, the
differences in DC between the experimental groups were
statistically significant as the linear mixed-effects model evaluates
differences on an individual-sample base. While DC varied from
specimen to specimen, the difference in top-bottom DC within one
sample was more consistent. The release from the three
composites is presented in Fig. 2 for the five experimental groups.
The highest release was recorded for BisGMA during the first week
for all the three composites in the five experimental groups, after
which it substantially dropped. TEGDMA was eluted almost as
much as BisGMA from the conventional flowable composite Filtek
Supreme XTE Flowable, when it was applied following both the
layer- and bulk-filling technique. UDMA was eluted at a relatively
steady rate from Filtek Z250 and Filtek Bulk Fill Flowable. No
UDMA was released from Filtek Supreme XTE Flowable, which
corresponds to the composition retrieved from the technical

Fig. 1. Graph representing the degree of conversion (DC in%) for the three composites measured at the top and bottom surface of the 4-mm high cylinder specimens for the
five experimental groups. The thick horizontal line in the box represents the median DC; the boxes represent the first quartile (Q1) to the third quartile (Q3) the whiskers
represent the lower and the upper quartile and the dark circle represent the mean value. Statistical significant differences in DC are indicated by asterisks. The difference
between DC top and bottom are indicated in percentage.

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Fig. 2. Graph representing the release (mmol) of the different monomers from the three composites as observed during 6 weeks for the five experimental groups.

information provided by the manufacturer. The release of BisEMA
(6) was similar to that of UDMA, with no BisEMA(6) released from
the flowable composite. Finally, very peculiar was the release of
BisPMA that markedly increased up to the fourth week for the two
flowable composites applied following both the layer- and bulk-
filling technique.
All composites tested released BPA. As expected, however, the
amount of BPA released was much lower than that of the other
monomers, this irrespective of the composite and filling technique
(Fig. 3). A steep reduction in release was noted after 1 week with a
more gradual decrease after 2 weeks. After 6 weeks, the release
BPA was very close to quantification limit (about 0.56 nmol).
The total monomer elution after 6 weeks is presented in Table 5.
The conventional paste-like composite Filtek Z250 Universal
released significantly fewer monomers than the other two
composites tested. A significantly higher total monomer elution
was recorded for both the flowable composites, irrespective of the
application method (bulk-filling or layer-filling technique). No
correlation was observed between total monomer elution and DC
at the top surface (r2 = 0.083, p = 0.042) nor at the bottom surface
(r2 = 0.527, p < 0.001) (Fig. 4).

4. Discussion

In this study, the release of monomers and the DC of three types

Fig. 3. Graph representing the release of Bisphenol A (BPA) as released from the
three composites during 6 weeks for the five experimental groups.
of composites from the same manufacturer were investigated. In
spite of their relatively similar monomer compositions, the total
monomer release varied significantly depending on the composite
type. The total monomer elution for the conventional paste-like
composite (Filtek Z250 Universal) was significantly lower than for
the conventional flowable (Filtek Supreme XTE Flowable) and the
bulk-fill flowable (Filtek Bulk Fill Flowable) composite. Bulk-filling
of the two flowable composites resulted in a significantly lower DC
at 4-mm depth (specimen bottom surface) and a significantly
higher overall release. The null hypotheses that (1) there was no
difference in DC and release for the three different composite
types, and (2) that the application technique (the layer-versus
bulk-filling technique) did not influence DC and release, must thus
be rejected.
The DC was measured using mRaman, a spectroscopic
technique often used for the determination of the polymerization

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Table 5
Total monomer elution after 6 weeks presented in mmol and in percentage (between brackets).

Filtek Z250 Universal Filtek Supreme XTE Flowable Filtek Bulk Fill Flowable

Layer-filled Layer-filled Bulk-filled Layer-filled Bulk-filled

BisGMA 0.162 (31.8) 0.199 (26.4) 0.382 (35.7) 0.213 (27.4) 0.320 (27.5)
BisEMA(6) 0.111 (21.8) 0 (0) <0.001 (0.1) 0.093 (12.0) 0.118 (10.2)
BisPMA 0 (0) 0.410 (54.5) 0.419 (39.0) 0.254 (32.7) 0.421 (36.2)
UDMA 0.200 (39.2) 0 (0) 0 (0) 0.210 (26.9) 0.299 (25.7)
TEGDMA 0.036 (7.1) 0.144 (19.1) 0.270 (25.2) 0.007 (0.9) 0.004 (0.3)
BPA <0.001 (0.1) <0.001 (0.1) <0.001 (0.1) <0.001 (0.1) <0.001 (0.1)
Total1 0.510  0.018a 0.754  0.033b 1.073  0.043c 0.777  0.022b 1.162  0.030d
1
Difference in superscript letter for the total monomer elution refers to a statistically significant difference.

degree by quantification of the amount of unreacted vinyl groups
[19–21]. In contrast to FTIR, mRaman does not require specific
specimen preparation and allows a non-destructive analysis. The
DC of the tested composites varied between 60 and 70% in all
experimental groups, which is the normal range for filled dental
composites [22]. DC measured at the top and bottom surface were
significantly different for all tested groups, even though the
difference in top-bottom DC was sometimes very small (<2%)
(Fig. 1). The statistical method used, mixed-effects model,
compares statistical differences on an individual-sample base,
which explains its high sensitivity. Nevertheless, in particular a
bulk filling application technique did result in clinically relevant
lower DCs at the bottom of the composite samples. This should
primarily be explained by the reduced curing time (20 s for bulk
filling technique versus 2  20 s for incremental technique, as per
instructions of the manufacturer). A reduction in DC of 7.3% and
4.2% for Filtek Supreme XTE Flowable (3M ESPE) and Filtek Bulk Fill
Flowable (3M ESPE), respectively, was recorded. As criterion for
sufficient polymerization, the cure at depth is considered adequate
if DC measured at the bottom reaches at least 90% of the maximum
DC measured at the top surface [10]. Strictly applying this
guideline, an adequate cure at 4-mm depth was achieved by the
flowable bulk-fill composite (Filtek Bulk Fill Flowable: 92%), but
not by the conventional flowable composite (Filtek Supreme XTE
Flowable: 89%). As the monomer composition and the basic
handling characteristics of the flowable and bulk-fill composite
were alike, the (rather slight) difference in depth of cure must most
likely be attributed to the larger and more translucent filler
fraction within Filtek Bulk Fill Flowable [23]. The slight increase in
top-bottom DC for Filtek Z250 and Filtek Bulk Fill when applied
incrementally must be attributed to increased curing time (40 s
instead of 20 s).
LC/MS–MS was used to determine the release of components,
since this technique is more accurate and has a lower detection
limit than high-performance liquid chromatography (HPLC) [24].
To analyze high molecular-weight monomers, such as BisGMA,
BisEMA(6), BisPMA and UDMA, gas chromatography combined
with mass spectrometry (GC/MS) is also not suitable. [25]
Moreover, using GC/MS, large monomers might decompose into
smaller molecules that affect the analysis, which was shown before
for UDMA that decomposed to hydroxyethyl methacrylate (HEMA)
[26]. Absolute ethanol was employed as elution medium, which
should be regarded as a worse-case scenario. Organic solvents have
the ability to penetrate the polymer network, allowing it to swell
and so facilitating the liberation of unreacted and leachable
monomers [27]. Saliva would be a more clinically relevant elution
medium, but it contains a complex mixture of organic and
inorganic ingredients that complicate accurate chemical assess-
ment of monomer elution [28]. Nevertheless, for some monomers,
such as TEGDMA [29] and UDMA [30], similar amounts were found
to be released from composite in saliva and organic solvents. For
other monomers, such as BisGMA [29] and BPA [30], the release in
organic solvents is much higher depending on the hydrophobicity
of the compounds.
Monomer elution depends on the molecular characteristics of
the monomers and the whole material composition [28]. To
monitor the kinetics of monomer release, elution should be
checked at several time points, which is not frequently done
[14,15,17,29,31], One can opt to take a sample of the eluate every
day [32], or one can refresh the solvent frequently as in this study.
Refreshing the solvent may be more clinically relevant, since it

Fig. 4. Graphs presenting the correlation analysis between DC at the specimen top and bottom surface and the absolute monomer elution during 6 weeks for the five
experimental groups.

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1454 P. Pongprueksa et al. / Journal of Dentistry 43 (2015) 1448–1455
simulates to some extent the saliva flow in the mouth. In general,
we observed a release of BisGMA and TEGDMA in an enhanced
amount during the first week, but their release considerably
dropped from the second week on. In contrast, BisPMA and UDMA
elution was relatively stable over time, or even continued to
increase from the beginning to the fourth week and then gradually
decreased towards the six-week time point. BisEMA(6) eluted at a
constant rate during the whole experiment. Thus, the monomer
composition of the eluate changed from mainly BisGMA/TEGDMA
to mainly BisPMA over time. This phenomenon may be caused by
saturation of the solvent with BisGMA/TEGDMA during the first
week, hampering the elution of BisPMA and UDMA. In addition,
pores created by elution of smaller monomers (like TEGDMA) may
facilitate the release of larger and more hydrophobic monomers
(like BisPMA) over time. More research is necessary to evaluate
potential toxicological implications of these findings.
BisEMA(6) eluted in a constant concentration over time but at
lower elution concentrations than BisGMA for Filtek
Z250 Universal and Filtek Bulk Fill Flowable. This could be
attributed to compositional differences, or to different elution
kinetics of BisEMA(6) and BisGMA. However, since the exact
proportion of the monomers in the composites is not disclosed by
the manufacturer, it is difficult to determine the exact reason.
BisEMA(6) has been used to substitute the large amounts of
TEGDMA present in the conventional paste-like (hybrid) compos-
ite Z100 in order to reduce polymerization shrinkage of the
successor composite Filtek Z250.
BisPMA is a monomer typically used in composites from 3 M
ESPE, and has been used to substitute the high content of the main
dimethacrylate monomers, such as BisGMA in composite. Its linear
structure of BisPMA is very similar to that of BisGMA, except that
there are two oxygens fewer than in BisGMA [33]. Its lower
molecular weight may explain its release at relatively high
concentrations over longer periods compared to BisGMA. Even
though this monomer showed a lower allergenic potential than
other dimethacrylate monomers in previous animal [34] and
human (patch test) research [35], more research should be
dedicated to its degradation and potential toxicity of the resultant
degradation products given the longer release period of BisPMA as
compared to that of BisGMA and TEGDMA (Fig. 2).
UDMA has been used to replace TEGDMA in the conventional
paste like composite Filtek Z250 Universal as compared to its
predecessor Z100, this in a similar way as mentioned above for
BisEMA(6). UDMA’s elution kinetics was similar to that of BisPMA;
its release increased from the beginning and slowly decreased
towards the end of the six-week evaluation period. Noteworthy are
the reported concerns regarding the cytotoxicity of UDMA [36,37].
Moreover, UDMA has been described to degrade into HEMA [26],
which is known for its cytotoxic and potential genotoxic effects [5–
7].
Since all tested composites contained monomers with a
bisphenol A-core, such as BisGMA, BisPMA and BisEMA(6), it
was no surprise that all composites released small amounts of
Bisphenol A (BPA). It is known that composites may contain BPA as
a residue of the synthesis process of these monomers. BPA has been
a toxicity concern in dental material science for more than a
decade. Recently, the European Food Safety Authority (EFSA) has
lowered the tolerable daily intake (TDI) from 50 mg/kg/bw/day
(microgram/kilogram/body weight/day) to 4 mg/kg/bw/day [38].
Nevertheless, the maximum BPA elution measured in this study
(0.053 mg for Filtek Bulk Fill Flowable after 1 week) was still a
significant magnitude lower than the tolerable amount for 1 week
(28 mg/kg/bw/week). As the size of the presently used composite
blocks (5-mm diameter  4-mm depth) more or less reflects a large
single-tooth restoration (like a MOD restoration on a molar or a
veneer restoration on a front tooth), it is unlikely that even the
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placement of several of these restorations will result in a high
exposure to BPA.
The lowest total monomer elution was found for the
conventional paste-like composite Filtek Z250, which was
significantly lower than the total monomer elution measured in
any other experimental group (Table 5). This must in the first place
be attributed to the lower resin content (15–25 wt%) in a
conventional restorative composite as compared to that of Filtek
Supreme XTE Flowable and Filtek Bulk Fill Flowable. Application
following the bulk-filling technique increased the total monomer
elution by about 30%. For the conventional flowable composite
Filtek Supreme XTE Flowable not only the total monomer elution
increased, but also the proportional composition of the eluate
changed. When applied following the bulk-filling technique, the
amount of BisGMA and TEGDMA almost doubled as compared to
when applied following a layer-filling technique. The release of
BisPMA remained, however, alike irrespective of the filling
technique employed.
No correlation between DC and monomer release was found at
the top nor the bottom specimen surface (Fig. 4). However,
significantly higher monomer elution was measured for the
specimens prepared using the bulk-filling technique, this in
accordance with a significantly lower DC measured at the
specimen bottom surface. But even for these specimens, DC had
no predictive value in terms of monomer elution. This could be
ascribed to several factors. First, there was a high variability in DC.
In general, we observed that the differences in DC between the
specimen top and bottom surfaces were smaller than the overall
variability in DC between different specimens. Also in another
study on the correlation between DC and release of well-
polymerized specimens, similar results were obtained [17,39].
Only in studies with suboptimal polymerization (by reducing
curing time [9] or by changing the type and concentration of the
photo-initiator [40]), a clear correlation between DC and release
was found. Secondly, it is likely that other factors than DC also
influence monomer release, such as for example cross-link density,
but more research is necessary to evaluate these factors.
To conclude, monomer release highly depends on the type of
composite and the application method. The slightly reduced
degree of conversion at 4-mm depth resulted in a higher monomer
elution when the composite was applied following a bulk-fill
application method.
Acknowledgements
Drs. Pong Pongprueksa received a scholarship from the Royal
Thai Government supported by the Ministry of Sciences and
Technology (MOST). The project was financed, in part, by the
G.0496.10 grant of the Research Foundation-Flanders (FWO). ESPE
gratefully acknowledged for providing the composite and BisPMA.
We would like to thank Esstech for providing raw monomers used
for the analytical analyses.
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