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journalhomepage:www.intl.elsevierhealth.com/journals/dema

Acrylamides and methacrylamides as alternative monomers

for dental adhesives

Stéfani Becker Rodrigues a, Cesar Liberato Petzhold b, Douglas Gamba b,

Vicente Castelo Branco Leitune a, Fabrício Mezzomo Collares a,∗
a
Dental Materials Laboratory, School of Dentistry, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil
b
Organic Chemistry Department, Chemistry Institute, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil

ARTICLE INFO ABSTRACT

Article history: Objective. Synthesize and characterize a methacrylamide monomer for adhesive system and evaluate the
Received 16 April 2018 physicochemical properties of the adhesive resin.
Received in revised form 9 July 2018 Methods. The liquid methacrylamide monomer N,N ,N -(nitrilotris(ethane-2,1-dyil)tris(2-methylacrylamide)
Accepted 27 August 2018 Available online (TMA) was prepared by reaction of methacrylic anhydride and tris(2-aminoethyl)amine with 60% yields. The
xxx TMA structure was analyzed by 1H NMR, 13C NMR, ATR-FTIR and UHPLC-QTOF-MS. Experimental
adhesive resin con-taining bisphenol-A glycidyl methacrylate (BISGMA), 2-hydroxyethylacrylamide
Keywords: (HEAA), 2-hydroxyethylmethacrylate (HEMA) and TMA were formulated. Polymerization kinetics of neat
Organic synthesis TMA and experimental adhesive resin (TMA33%/HEAA66%, TMA50%/HEAA50%, TMA66%/HEAA33%,
Dental adhesives TMA50%/HEMA50%, BisGMA/HEAA/TMA and BisGMA/HEMA) were evaluated using Differential
Acrylamides Scanning Calorimetry. Physiochemical properties for Bis-GMA/HEAA/TMA and BisGMA/HEMA adhesives
Differential Scanning Calorimetry were evaluated by cytotoxicity, ultimate tensile strength (UTS), softening in solvent ( KHN), contact angle
Cytotoxicity (u), microtensile bond strength (mTBS) and failure analysis. A primer was also formulated with
Degree of conversion H2O/HEAA/AMPS (2-acrylamido-2-methylpropane sulfonic acid) and the pH value was verified and
compared to commercial primer.

Results. Adhesive resin with only HEAA and TMA (TMA33%/HEAA66%, TMA50%/HEAA50%,
TMA66%/HEAA33%) showed lower conversion and polymerization rate after 40 s of light activation.
Conversion up to 60% was found for BisGMA/HEAA/TMA and BisGMA/HEMA adhesive resin without
significant difference between groups, p > 0.05. Cytotoxicity, UTS, mTBS, KHN and u showed no statistical
difference, p > 0.05, between BisGMA/HEAA/TMA and BisGMA/HEMA adhesive resin.

Significance. In this study, the proposed synthetic route resulted in a tris(methacrylamide). A new primer
composed without acrylates or methacrylates was formulated for 3-step etch-and-rinse adhesive system
without the presence of HEMA monomer. Physicochemical

∗
Corresponding author at: Dental Materials Laboratory, School of Dentistry, Federal University of Rio Grande do Sul, Porto Alegre, Rua Ramiro Barcelos, 2492
Rio Branco, 90035-003 Porto Alegre, RS, Brazil.
E-mail address: fabricio.collares@ufrgs.br (F.M. Collares).
https://doi.org/10.1016/j.dental.2018.08.296
0109-5641/© 2018 Published by Elsevier Ltd on behalf of Academy of Dental Materials. All rights reserved.

Please cite this article in press as: Rodrigues SB, et al. Acrylamides and methacrylamides as alternative monomers for dental adhesives. Dent Mater (2018),
https://doi.org/10.1016/j.dental.2018.08.296
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properties and cell viability of BisGMA/HEAA/TMA adhesive resin represents an alternative

adhesive resin without HEMA monomer.
© 2018 Published by Elsevier Ltd on behalf of Academy of Dental Materials. All rights
reserved.

chemical properties of new 3-step etch-and-rinse adhesive

1. Introduction system with acrylamides.

The degradation of adhesive resin is related to the for-

mulation of dental adhesives with hydrophilic functional
2. Methodology
monomers and the presence of water. These adhesive sys-
tems are composed by monomers having hydroxyl, carboxyl,
and ester groups making them more prone to water sorp- 2.1. Monomer synthesis
tion, leading to the disruption of covalent bonds between
monomers of polymeric chain and, consequently, resin weight To a solution of tris(2-aminoethyl)amine (0.533 g; 3.41 mmol)
loss [1–5]. Therefore, resulting in permeable adhesive inter- and triethylamine (3.5 eq.) in 30 mL of anhydrous
face that also promotes lower elastic modulus of adhesive dichloromethane, a solution of methacrylic anhydride
resin and compromises the long-term of restoration treatment (3.5 eq.), in 10 mL of anhydrous dichloromethane, was added
[6]. dropwise in ice bath for 30 min, according to Fig. 1. After stir-
Methacrylates are the most usual monomers in adhesive ring for 16 h at room temperature, 10 mL of water was added,
systems due to chemical and mechanical properties [7–10]. and the aqueous phase was extracted with dichloromethane
However, when exposed to acidity or aqueous environment (3 × 5 mL). After drying the organic phase with anhydrous
the hydrolysis of ester groups occurs quickly, mainly for Na2SO4, the solvent was removed in a rotator evaporator at
hydroxyethyl methacrylate (HEMA) monomer [11,12]. Thus, room temperature protected from light. The crude product
some alternatives such methacrylamides or acrylamides was purified by column chromatography (Silica gel Si 70–230
monomers for HEMA were proposed [13–15]. Since carbamides Mesh), using ethyl acetate:methanol (99:1) as eluent to afford
are more stable hydrolytically due to the lower reactivity of the monomer N,N , N -(nitrilotris(ethane-2,1-dyil)tris(2-
carbonyl group when compared to the ester monomers their methylacrylamide) as a light yellow viscous liquid in 60%
1 13
hydrolysis occurs only in an extreme condition, such as a high yield. H NMR, C NMR, mid-IR, and Mass spectrum are
acid (pH <0.94) environment [7]. Furthermore, acrylamides shown in Appendix A.
1
bond with collagen fibrils by hydrogen bond could be another H NMR (300 MHz, CDCl3) ı (ppm): 6.83 (s, 3H); 5.71 (s, 3H);
advantage over methacrylates [16,17]. 5.29 (s, 3H); 3.39 (q, J = 5.6 Hz, 9H); 2.66 (t, J = 5.6 Hz, 6H), 1.92 (s,
Some acrylamides were synthesized and the final product 9H).
13
was a solid monomer that makes the dissolution with other C NMR (100 MHz, APT) ı (ppm): 168.9 (3); 139.3 (3); 119.7
monomers or solvents more difficult [7,18]. In the meantime, (3); 53.6 (3); 37.3 (3); 18.4 (3).
new routes were developed generating liquid bis(acrylamides)
as an alternative to methacrylate monomers in adhesive resin
2.2. Monomer characterization
as well as primers [7,15,18–30]. These liquid monomers, such
as N,N -diethyl-1,3-bis-(acrylamido)-propane (DEAAP), N,N - 2.2.1. Nuclear magnetic resonance (NMR)
dimethyl-1,3-bis-(acrylamido)-propane (DMAAP) and N,N - NMR measurements were recorded on a Bio Spin GmbH
dimethyl-1,6-bis-(acrylamido)-hexane (DMAAH) showed a 1 13
(Bruker Biospin, Rheinstetten, Germany), H: 400 MHz, C:
similar viscosity as commercial dimethacrylate diluents such
100 MHz, in CDCl3 containing tetramethylsilane (TMS) as
as triethyleneglycol dimethacrylamte (TEGDMA), were com-
an internal standard. The multiplicities were attributed as:
pletely miscible with hydrophilic or hydrophobic monomers
s = singlet; d = doublet; t = triplet; qt = quintuplet; dd = double
and show improved hydrolytic stability [15]. However, the
dublet; ddd = double double dublet e m = multiplet. The hydro-
N-disubstituted acrylamides present a lower reactivity in
gen assignments were attributed based on relative integral
radical homopolymerization compared to unsubstituted acry-
and coupling constant (J) in Hertz (Hz).
lamides. Therefore, new unsubstituted acrylamides and
methacrylamides should be established in the literature,
as the tris(methacrylamide). The addition of trifunctional 2.2.2. Attenuated total reflectance-Fourier transform
methacrylamide monomer, with high molecular weight, in infrared spectroscopy (ATR-FTIR)
the experimental comonomer blend could result in adhesive ATR-FTIR measurements were performed in the Bruker Alpha
resin with increased crosslink density, due to three C C bond. FTIR Spectrometer (Bruker Optics, Ettlingen, Germany). The
Consequently, trifunctional methacrylamide monomer could monomers were dispensed over a diamond crystal of Attenu-
result in an adhesive resin with high mechanical properties as ate total reflectance (ATR) accessory. A total of 64 scans were
collected from 400 cm− to 4000 cm− at a 4 cm− resolution
1 1 1
alternative adhesive resin without HEMA monomer [10,31,32].
Thus, the aim of this study was to synthesize and characterize for each specimen.
a tris(methacrylamide) monomer and evaluate the physico-

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2-methyl-propanesulfonic acid (AMPS). A commercial primer,

Table 1 – TMA homopolymer and experimental adhesive formulation
groups in wt%. Scotchbond Multi-Purpose (3M ESPE, St. Paul, MN, USA), was utilized
to compare with experimental primer.
Groups TMA HEAA BisGMA HEMA
TMA100% 100% – – –
2.3.1. pH analysis
TMA33%/HEAA66% 33.3% 66.7% – –
The pH of experimental and control primer was evaluated using a digital
TMA50%/HEAA50% 50% 50% – –
TMA66%/HEAA33% 66.7% 33.3% – – pHmeter (DM-23; Digicrom Analytical Ltda., São Paulo, SP, BR). Three
TMA50%/HEMA50% 50% – – 50% measurements were performed per group.
BisGMA/HEAA/TMA 10% 24% 66% –
BisGMA/HEMA – – 66.7% 33.3% 2.3.2. Degree of conversion and polymerization kinetics
TMA: N,N ,N -(nitrilotris(ethane-2,1-diyl))tris(2-methylacrylamide); The degree of conversion (DG) and rate of polymerization (Rp) of
HEAA: 2-hydroxyethyl acrylamide; BisGMA: Bisphenol A glycero- homopolymer TMA and resin adhesives (TMA 33.3%/HEAA 66.7%;
late; HEMA: 2-hydroxyethyl methacrylate.
TMA 50%/HEAA 50%; TMA 66.7%/HEAA 33.3%; TMA 50%/HEMA
50%; BisGMA/HEAA/TMA; BisGMA/HEMA) were evaluated via
differential scanning calorimetry (DSC; DSC-Q2000, TA Instrument Co.,
2.2.3. Ultra-high liquid chromatography quadrupole time of DE, USA) with a photocalorimetric accessory (PCA) according Rodrigues
flight mass spectrometry (UHPLC-QTOF-MS) et al. [31]. The intensity and wavelength of the PCA were adjusted to 100
High-resolution mass spectra were obtained with a Q-TOF Micro 2
mW/cm and 390–500 nm, respectively. Three samples per group
instrument (Impact II, Bruker, Bremen, Germany) in electrospray
ionization positive (ES+) mode. A Q-TOF system was used to separate (approxi-mately 12 mg) were polymerized in open aluminum DSC pans
the analytes of interest. Shim-pack XR-ODS III column (50 mm × 2 mm and covered with a quartz lid. An additional empty aluminum pan was
× 1.6 mm) was used to separate the analytes in isocratic mode with the used as a control. The lamp was switched on for 3/10 min and heat flow
mobile phase 40% acetonitrile (0.1% formic acid):60% water (0.1% was recorded. The analysis was per-formed with a nitrogen flow rate of 50
formic acid) (LC, Nexera ×2, Shimadzu, Tokyo, Japan). The flow rate mL/min. Assuming that the value of the heat involved is proportional to
the reacted molar amount, the degree of conversion was deter-mined
was 0.4 mL/min and the column temperature was 35 ◦ C. The opti-mal
according to the following Eq. (1):
MS parameters were as following: capillary voltage 4500 V, source
temperature 200 ◦ C, end plate offset voltage 500 V, mass range (m/z) of
150–700 and calibration with sodium formate.

a (mol%) = 100 [M]0 − [M] = 100Q, (1)

2.3. Adhesive formulation
[M]0 ( Hp,0/ Hp)Qtot
Six experimental adhesive resins were produced in order to analyze the
Hp,o (kJ mol− ) corresponds to the heat of polymerization for a
influence of N,N ,N -(nitrilotris(ethane-2,1-diyl))tris(2-methylacrylamide) 1

total conversion, Hp (kJ mol− ) is the heat of poly-merization obtained by

incorporation at adhesive resins, Table 1. The co-initiation system was
composed by 1.0 mol% camphorquinone (CQ) and 1.0 mol% ethyl 4-
dimethylaminobenzoate (EDAB) for all groups. Reagents were purchased
from Sigma-Aldrich (St. Louis, MO, USA). The com-ponents were
weighed using an analytical balance (AUW220D, Shimadzu, Tokyo,
Japan), and were mixed and ultrasonicated (Cristófoli, Campo Mourão,
PR, BR). To perform monomer photoactivation, a light-emitting diode
2
unit (Radii Cal, SDI Ltd., AU), at an intensity value of 1200 mW/cm ,
1
the apparent area of the curve that corresponds to the total heat of reaction
Qtot (J g− ), and Q cor-responds to the heat released as partial area under
1
the curve for a time, t. The Hp,o value for a double bond of methacry-lates
has been reported to be −56 J mol− [33], for acrylamides, −82.9 J mol−
1 1
and for methacrylamides, −35.2 J mol− [34]. The rate of polymerization
1
(Rp) is proportional to the heat flow released in the isotherm as a function
of irradiation time (t). Thus, Rp (mmol g− s− ) at any point during the
1 1

was used. reaction can be derived from the heat flow using the Hp,o of the monomer
according to the following Eq. (2):
One experimental primer with acrylamide was formulated with 54.9%
of water, 45% of HEAA and 0.1% of 2-acrylamido-

Fig. 1 – Reaction scheme of the tris(methacrylamide) synthesis. Et3N-triethylamine; CH2Cl2-dichloromethane; r.t.-room temperature.

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Rp = (dH/dt).(M/ Hp,0.n),
where dH/dt is the heat flow in J mol− s− , M is the concen-
1 1
tration of the monomer, and n is the number of double bonds
per molecule of monomer.
2.4. Mechanical properties of experimental adhesives
resin: BisGMA/HEMA and BisGMA/HEAA/TMA
2.4.1. Softening in solvent
Five samples per group, BisGMA/HEMA, and BisGMA/HEAA/
TMA, were prepared (5.0 ± 0.5 mm diameter and 1.0 ± 0.2 mm
thickness) in a polyvinylsiloxane-based mold. Samples were
subsequently embedded in an acrylic resin and polished
(Model 3v, Arotec, Cotia, SP, BR) with a felt disc that was sat-
urated with an alumina suspension (Alumina, 6 mm, Arotec,
Cotia, SP, BR). Five indentations (10 g/5 s) 100 mm apart from
each other were made in each specimen using a digital hard-
ness Knoop tester (HMV 2, Shimadzu, Tokyo, Japan). The
samples were immersed in a solution of ethanol:water (70:30)
for 2 h. Before and after the immersion period, samples were
submitted to the hardness test (KHN1 and KHN2) and the per-
centual hardness reduction was recorded [35].
2.4.2. Ultimate tensile strength
Five samples per group, BisGMA/HEMA and Bis-
GMA/HEAA/TMA, were prepared in a metallic matrix with an
hourglass design measuring 8 mm long, 2 mm wide, and 1 mm
2
thick, with a cross-sectional area of 1 mm . The adhesives
were light cured for 20 s on each side. The samples were
fixed with a cyanoacrylate adhesive in a metallic device,
and the tests were evaluated in a universal testing machine
(EZ-SX Series; Shimadzu, Tokyo, Japan) at a crosshead speed
of 1 mm/min until failure and the values were expressed in
MPa [36].
2.4.3. Preparation of teeth to contact angle and
microtensile bond strength test
The bovine permanent teeth were extracted, cleaned of
organic debris and stored in distilled water at 4 ◦ C. The buccal
enamel was removed using a water-cooled, low-speed dia-
mond saw to expose the superficial dentine. The smear layer
was produced with 600-grit SiC paper for 30 s in running water.
2.4.4. Contact angle measurement
Five dentin surfaces were polished with 600-grit silicon car-
bide abrasive paper under the wet condition for 30 s, etched
with 37% phosphoric acid for 15 s, washed and dried. A drop
(3 mL) of control (Scotchbond multi-purpose, 3M ESPE, St. Paul,
MN, USA) and experimental (water/HEAA/AMPS) primer was
placed on dentin surface and the static contact angle (u)
between the droplet and the solid surface. After that, primers
were vigorously applied in dentin, and the solvent was evap-
orated for 10 s. A drop of adhesive (3 mL), BisGMA/HEMA and
BisGMA/HEAA/TMA, was placed over dentin with primer and
the contact angle was measured. The samples were analyzed
by an optical tensiometer (Theta, Biolin Scientific, Stock-
holm, Sweden) to measure the mean contact angle (u) to
primer/adhesive and to dentin surface. The parameters used
such as the drop out size, the drop rate, the displacement rate
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(2) and the speed dispersion of the liquids were according to pre-
vious study [37]. In 20 s the test was performed and after 10 s
the mean contact angle was recorded.
2.4.5. Microtensile bond strength ( TBS) and failure
pattern analysis
Twenty dentin surfaces per group, BisGMA/HEMA, and Bis-
GMA/HEAA/TMA, were polished with 600-grit silicon carbide
abrasive paper under the wet condition for 30 s, etched
with phosphoric acid for 15 s, rinsed and dried. Primer
(commercial-Scotchbond multi-purpose or experimental-
H2O/HEAA/AMPS) was vigorously applied for 20 s, and the
solvent was evaporated for 20 s. The adhesives (BisGMA/HEMA
and BisGMA/HEAA/TMA) were applied and photoactivated
for 20 s using a light-emitting diode (Radii cal, SDI, Bayswater,
AU). A composite build-up was performed (Z350, 3M ESPE, St.
Paul, USA) in two increments of 2 mm. After storage in dis-
tilled water at 37 ◦ C for 24 h, the teeth were sectioned in four
2
beams (approximately area of 0.5 mm ) with a slow-speed
saw. The beams were fixed with a cyanoacrylate adhesive in a
metallic device to be submitted to a universal testing machine
(EZ-SX Series; Shimadzu, Tokyo, Japan) at a crosshead speed
of 1 mm/min until failure with values expressed in MPa.
Fractographic failure mode was evaluated with a stereomicro-
scope (HMV2; Shimadzu, Tokyo, Japan) at 10× magnification
and classified as adhesive/mixed or cohesive in dentin/resin
composite.
2.4.6. Cytotoxicity sulforhodamine B (SRB) colorimetric
assay
Primary pulp fibroblasts were derived from two intact human
third molars with incomplete root formation of different
patients without systemic health problems that agreed to
donate tooth for this study and signed the informed con-
sent. For the cytotoxicity assay, the eluate was prepared
immersing the specimen (3 mm diameter × 1 mm thickness)
for BisGMA/HEMA and BisGMA/HEAA/TMA resin adhesives
and for the control group in 1 mL of the medium during 72 h.
3
Cells were seeded in triplicate at a concentration of 5 × 10
per well in 96 well plates, and then treated with 100 mL of
eluate. After 72 h cells were fixed with a concentration of
10% trichloroacetic acid (TCA), they were incubated at 4 ◦ C
for 1 h and subsequently were washed 6 times under run-
ning water and dried at room temperature. Sulforhodamide
B (SRB, Sigma-Aldrich, St. Louis, USA) at 0.4% was added to
stain the cells and the plate was incubated for 30 min at room
temperature. The plate was washed 4 times with 1% acetic
acid to remove unbound excess dye and allowed to dry com-
pletely at room temperature. Trizma solution was added and
the plate incubated for 1 h to allow complete solubilization of
the dye. At the end of this process, the microplates were read
at 560 nm. Cell viability was normalized against the viability
of cells in wells without treatment according to previous study
[38]. Six samples were used to test cell viability and tests were
performed in triplicate
2.5. Statistical analysis
The normality of the data was evaluated using the
Shapiro–Wilk test. Statistical analysis for the degree of
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Fig. 2 – Photopolymerization of homopolymer TMA and TMA/HEAA adhesive systems with different weight proportions by DSC-PCA. TMA100% –
100% of TMA; TMA33%/HEAA66% – 33.3% of TMA + 66.7% of HEAA; TMA50%/HEAA50% – 50% of TMA + 50% of HEAA;
TMA66%/HEAA33% – 66.7% of TMA + 33.3% of HEAA; TMA50%/HEMA50% – 50% of TMA + 50% of HEMA.

Fig. 3 – Degree of conversion and polymerization rate of experimental adhesive resins BisGMA/HEMA and BisGMA/HEAA/TMA.
BisGMA/HEAA/TMA – 66% of BisGMA + 24% of HEAA + 10% of TMA; BisGMA/HEMA – 66.7% of BisGMA + 33.3% of HEMA.

Table 2 – Degree of Conversion (DC) and Rp of the TMA homopolymer and experimental adhesive resins.

Rp (mmol g− s− )
1 1
Groups DC (%)
DC
DC tmax (s) t40s Rpmax tmax (s)
D C
TMA100% 35.6 (±1.69) 412 22.3 (±1.28) 3.2 10.4
TMA33%/HEAA66% 71.1 (±4.57)ABC 556 15.1 (±0.75)CD 2.1 41.1
TMA50%/HEAA50% 82.0 (±3.54)A 1042 10.6 (±0.84)D 1.3 10.9
C B
TMA66%/HEAA33% 60.0 (±2.61) 219 39.0 (±0.9) 5.2 7.6
TMA50%/HEMA50% 78.0 (±1.70)A 409 21.6 (±0.60)C 2.0 40
BisGMA/HEMA 63.7 (±4.04)AB 41 62.5 (±2.65)A 11.3 7.7
69.0(±14.5)ABC
A
BisGMA/HEAA/TMA 292 60.5 (±8.63) 8.4 3.3
TMA: tris(methacrylamide); HEAA:2-hydroxyethyl acrylamide; HEMA:2-hydroxyethyl methacrylate; BisGMA: Bisphenol A glycerolate dimethacrylate. Different capital
letter indicates statistical difference in same column (p < 0.001).

1 refer to N H bond (3300 cm− ); 2—C O peak (1660 cm− ); 3—C C

conversion was performed using One-way ANOVA and for contact angle, 1 1
pH, ultimate tensile strength, microtensile bond strength analysis, KHN%,
peak (1610 cm− ) and 4—C N peak (1520 cm− ). The reaction was
1 1
KHN1 and cell viability were performed using Student’s t-test, with 0.05 1
confirmed by IR spectrum. In the H NMR spectrum, the vinylic
of the level of significance. For comparison between initial and final
13
micro-hardness (KHN1 and KHN2) a paired Student’s t-test was used. hydrogens signals were observed at 5.29 and 5.71 ppm and in the C
2
NMR spectrum signals of the sp -hybridized C-atoms of the acrylamide
double bonds appeared at 119.7 and 139.3 ppm. The exact m/z value of
TMA, 351.2385 g/mol, was determined by UHPLC-QTOF-MS and the
3. Results structure confirmed by comparison with the the-oretical isotopic profile.
Mass error (err[ppm]) and isotopic profile (mSigma) between the
3.1. Synthesis and characterization of monomers theoretical profile and the sam-ple were calculated, 1.7 ppm and 7.4
1 13
respectively. H NMR, C NMR, mid-IR, and Mass spectrum are shown
Tris(methacrylamide) was prepared in one-step reaction via nucleophilic in Appendix (Figs. A1, A2 and A3).
substitution of methacrylic anhydride and tris(2-aminoethyl)amine with a
yield of 61%, Fig. 1. The main peaks in the IR spectrum were
demonstrated in Appendix when peak

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3.2. Physicochemical properties of dental adhesives 1

The polymerization behavior of monomer TMA and the adhesive resin
was shown in Figs. 2 and 3 and Table 2. TMA50%/HEAA50% adhesive
resin showed the highest degree of conversion, despite the prolonged
period of light emission (1042 s). When the DC was compared after 40 s
irradiation time the higher value was 62.5% for BisGMA/HEMA adhesive
resin following by 60.5% for BisGMA/HEAA/TMA and no difference
was found, p = 0.804. The higher Rpmax, 11.3 mmol g− s− , was found
1 1
anhydride and tris(2-aminoethyl)amine and the structure con-firmed by H
13
and C NMR, FTIR and UHPLC-QTOF-MS. Kinetics of TMA
photopolymerization and adhesives resin with dif-ferent weight
proportions of TMA, HEAA, and BisGMA were investigated. The
physicochemical properties of the adhesive resins were also evaluated.
The addition of a trifunctional methacrylamide monomer, TMA, showed
similar physico-chemical properties of formulated adhesive resins without
HEMA monomer.

for BisGMA/HEMA adhesive resin and the lower value (2.0 mmol g−
1
s− ) for TMA50%/HEMA50% adhesive resin.
1 TMA by N-acylation reactions between methacryloyl chloride and tris(2-
The results of pH of experimental
(H2O/HEAA/AMPS) and Scotchbond primer was
4.0 respectively, p < 0.001. The contact angle of
(H2O/HEAA/AMPS and Scotchbond) and adhesive resin (Bis-
GMA/HEMA and BisGMA/HEAA/TMA) showed in Table 3. Significant
differences between the contact angle values for primers were observed,
33.5 (±4.8) for Scotchbond primer and 18.5 (±5.4) for
H2O/HEAA/AMPS, (p = 0.002). For adhesive resins, the addition of
HEAA and TMA in BisGMA/HEAA/TMA group showed no significant
difference with BisGMA/HEMA adhesive resin, p = 0.387 (Fig. 4).
Knoop hardness and the percentage of softening in solvent was shown
in Table 3. The BisGMA/HEMA resin had higher KHN1 and KHN2
values than the BisGMA/HEAA/TMA adhesive resins (p < 0.001 and p =
0.001). Significant differences between the initial and final hardness
values for each resin were observed (p < 0.05). Addition of HEAA and
TMA showed no significant difference between percentage differences (
KHN) (p = 0.065).
The ultimate tensile bond strength for BisGMA/HEMA and
BisGMA/HEAA/TMA adhesives resin values were not statisti-cally
significant (p = 0.110), Table 3. The immediate mTBS was not
statistically significant between groups (p = 0.402) and the mixed failure
was predominant. Cytotoxicity assay resulted in 96.35% (±5.0) and 93.9%
(±1.0) of cell viability for Bis-GMA/HEMA and BisGMA/HEAA/TMA
group, respectively, and values were not statistically significant, p = 0.25.
4. Discussion
Patras et al. firstly described the synthesis and characteri-zation of
aminoethyl)amine resulting in a solid com-pound with m.p. of 141–142 ◦
primer C [39]. However, no investigation about the polymerization of this
2.7 and monomer was presented it has not been used for adhesives formulations.
primers Unexpectedly, the TMA synthesized from methacrylic anhydride was
liquid. Even after column purification and exhaustive attempts to
crystallize, the product remains liquid and, the characteriza-tion confirmed
the structure of the monomer (Fig. A3).
The kinetics behavior in this study investigated the
homopolymerization of TMA and its copolymerization with HEAA in
different weight proportions. As TMA present three polymerizable
methacrylamide groups, high molecular weight and initial high viscosity,
the homopolymer presented a lower degree of conversion (22.3%) due to
the rigid crosslinked poly-mer chain structure formed (vitrification
process) at earlier stages of the polymerization and lower mobility
between chains. Although HEAA homopolymer presented high R p (24.4
mmol g− s− ) and DCt40s (79%) [31], when TMA was added to the
1 1
formulation (TMA/HEAA) in all concentrations, the copolymerization rate
decreased and higher conversion was only reached after longer light
exposure time, Table 2 and Figs. 2 and 3. It is known that
methacrylamides present rela-tively low reactivity in homopolymerization
due to the steric hindrance of the alkyl substitutes and the onset of
vitrifica-tion at low degrees of conversion [40]. As shown in Fig. 4, when
TMA and HEAA are simultaneously copolymerized, two reactive radical
species could be formed in the initiation step: one resulting in a more
stable tertiary radical (Fig. 4-a) and other in a less stable secondary radical
(Fig. 4-b). Then, in the propagation step, the methacrylamide radical will
react pref-erentially with TMA monomer than acrylamide monomer. In
this way, the higher initial concentration of TMA results in a tapered
copolymer presenting a block of TMA homopolymer

In this study, trimethacryloyl-tris(2-aminoethyl)amine (TMA) was

prepared in the one-step reaction between methacrylic

Table 3 – Results of contact angle ( ) of the experimental adhesives, hardness before (KHN1) and after the immersion in solvents (KHN2), the
percentage of Knoop hardness decrease after ethanol/water immersion ( KHN) and ultimate bond strength (UTS).

Groups u KHN1 KHN2 KHN (%) UTS mTBS Failure pattern

A:12
M:43
A Aa Ab
BISGMA/HEMA 39.5 (±9.0) 18.3 (±0.3) 8.0 (±1.4) 56.0 (±7.6)A 67.7 (±5.8)A 57.0 (±14.1)A CD:20
CC:5
A:9
A Ba Bb
BISGMA/HEAA/TMA 46.7 (±15.0) 12.8 (±0.8) 4.6 (±0.7) 64.4 (±4.6)A 60.5 (±7.7)A 53.1 (±15.4)A M:67
CD:4

A: adhesive failure; M: mix failure; CD: cohesive dentin failure; CC: cohesive composite failure. Different small letter indicates statistical difference in same row (p <
0.05). Different capital letter indicates statistical difference in same column (p < 0.05).

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(Fig. 4-c) followed by a block of HEAA homopolymer, separated by an
intermediary random copolymer segment (Fig. 4-d). This behavior is
probably due to the stability of the activated monomer and a brief time
decreased monomers mobility due to the environment vitrification [9,41].
When the con-centration of HEAA increases, increases the probability of
the formation of acrylamide radical, which also reacts preferen-tially with
the TMA monomer favoring the copolymerization. However, as the TMA
was consumed, the polymerization of HEAA proceeds and a copolymer
presenting an initial random copolymer segment followed by an HEAA
block copolymer was obtained. Therefore, the long block of HEAA
formed lead to decreased mechanical properties similarly of already
showed in a study for a GBisGMA/HEAA adhesive resin [31].
Copolymerization kinetics of an adhesive resin consti-tuted of TMA
and HEMA (weight proportion 50:50) was also investigated (Table 2).
Although the homopolymerization rate of HEAA is higher than HEMA
[31], the copolymerization of HEMA with TMA is faster, since both
monomers result in propagating tertiary radicals. As consequence, a ran-
dom polymer was obtained. However, the copolymerization kinetics
remains beneath the classical BisGMA/HEMA sys-tem. Thus, all
adhesive resin composed with acrylamides and/or tris(methacrylamide)
showed a lower degree of con-version when exposed at light activation for
40 s and therefore were not test mechanical properties. Neverthe-less, the
structure of TMA provides an increase at crosslink density of
BisGMA/HEAA adhesive resin and, consequently,

Fig. 4 – Scheme of polymerization process between TMA and HEAA.

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the mechanical properties. Meanwhile, Rpmax of the sys-tem
BisGMA/HEAA/TMA (8.4 mmol g− s− ) is lower than of the
1 1
methacrylic esters BisGMA/HEMA (11.3 mmol g− s− ).
1 1
BisGMA/HEAA/TMA exhibited a delay in initial stages of
photopolymerization [25] but reaching a similar degree of con-version
after 40 s irradiation.
In this study, the crosslink density of BisGMA/HEAA/TMA adhesive
resin was increased due to the chemical structure of TMA, when
compared to acrylamide adhesive resin in a previous study [31]. The
radical activated methacrylamide has a preference to copolymerize with
BisGMA promoting a rigid polymer chain. This difference in polymer
crosslink density was noted in the results of initial Knoop hardness
(KHN1) and mechanical properties of BisGMA/HEAA/TMA which are
higher than KHN1 and mechanical properties of adhesive resin without
TMA monomer in the adhesive resin (66.6%Bis-GMA/33.3%HEAA) of a
previous study [31].
Even if compared to the BisGMA/HEAA/TMA and Bis-GMA/HEMA
adhesive resin, KHN1 and mechanical properties values are significantly
lower, p < 0.05, despite not show-ing the difference in the degree of
conversion in 40 s and KHN%. Although polymeric degradation and
leaching of unreacted monomers can lead to cytotoxic, in this study the
experimental adhesives showed no cytotoxic effect in pri-mary pulp
fibroblasts in 72 h. In the same way, the high degree of conversion and the
similar contact angle allowed the bond strength between the
dentin/adhesive interface with BisGMA/HEAA/TMA adhesive resin
without a difference for BisGMA/HEMA adhesive resin, p < 0.05.
A commercial primer was used for the bond strength of
BisGMA/HEMA adhesive resin and, for BisGMA/HEAA/TMA adhesive
resin, a new primer was formulated to evaluated an interface resin without
HEMA monomer. This primer was characterized by pH and contact angle
and presented a lower pH and greater wettability than the commercial one.
The
difference was due to the composition of the experimental primer where
more water is present which lowers the con-tact angle with dentin. High
amount of water is due to the addition of AMPS monomer, this monomer
is a strong acid and just 0.01 wt% could be added to the 3-step etch-and-
rinse adhesive system. Due to the increased crosslink density with the
addition of TMA and a more susceptibility to hydrolytic degradation of
methacrylates than to methacrylamides and acrylamides [11], it is
expected that in 1 year of water stor-age a higher degradation of
BisGMA/HEMA groups interfaces leading to lower bond strength.
With the limitations of these study, the formulation of a conventional
3-step etch-and-rinse adhesive system with tris(methacrylamide)
monomer, BisGMA/HEAA/TMA, repre-sents an alternative to a new
adhesive system without HEMA monomer, without altering degree of
conversion, contact angle, softening in solvent, ultimate bond strength,
immedi-ately microtensile bond strength, failure pattern and cellular
viability. Future studies related to the long-term degradation of these
adhesive interfaces will be performed to confirm this possibility. As well
as, this monomer is an alternative to using in self-etch adhesive.
Acknowledgments
The authors gratefully acknowledge CAPES (Coordenac¸ ão de Aperfeic¸
oamento de Pessoal de Nível Superior) for the schol-arship (S.B.R.), Julio
C.P. Vaghetti of the LAMAT (Laboratório Multiusuário de Análise
Térmica) and Alexsandro Dallegrave of the UHPLC-QTOF-MS. This
research did not receive any spe-cific grant from funding agencies in the
public, commercial, or not-for-profit sectors.
Appendix A.

Fig. A1 – FTIR-ATR spectrum of TMA. 1—N−H (3300 cm− ); 2—C O (1660 cm− ); 3—C C (1610 cm− ); 4—C–N (1520 cm− ).
1 1 1 1

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1 13
Fig. A2 – H NMR (a) and C NMR (b) of tris(methacrylamide) monomer. H NMR (300 MHz, CDCl3) = 6.83 (ls, 3H); 5.71 (s, 3H); 5.29 (s, 3H); 3.39
13
(q, J = 5.6 Hz, 9H); 2.66 (t, J = 5.6 Hz, 6H), 1.92 (s, 9H). C NMR (100 MHz, APT) = 168.9 (3); 139.3 (3); 119.7 (3); 53.6 (3); 37.3 (3); 18.4 (3).

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Fig. A3 – Mass spectrum of monomer TMA (a) and theoretical isotopic profile (b).

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