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Evaluation of Antibio LM and Mechanical Properties of New Nanocomposites Based On Acrylic Resins and Silver Vanadate Nanoparticles
Evaluation of Antibio LM and Mechanical Properties of New Nanocomposites Based On Acrylic Resins and Silver Vanadate Nanoparticles
PII: S0003-9969(16)30059-0
DOI: http://dx.doi.org/doi:10.1016/j.archoralbio.2016.03.002
Reference: AOB 3565
Please cite this article as: Castro Denise T, Valente Mariana LC, Silva Cláudia HL,
Watanabe Evandro, Siqueira Renato L, Schiavon Marco A, Alves Oswaldo Luiz, Reis
Andréa C.Evaluation of antibiofilm and mechanical properties of new nanocomposites
based on acrylic resins and silver vanadate nanoparticles.Archives of Oral Biology
http://dx.doi.org/10.1016/j.archoralbio.2016.03.002
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Evaluation of antibiofilm and mechanical properties of new nanocomposites based on acrylic
resins and silver vanadate nanoparticles
(a)
Postgraduate student, Department of Dental Materials and Prosthesis, School of Dentistry of
Ribeirão Preto, USP - University of São Paulo, Ribeirão Preto, SP, Brazil.
(b)
Professor, Department of Dental Materials and Prosthesis, School of Dentistry of Ribeirão Preto,
(c)
Postgraduate student, Postgraduate Program in Materials Science and Engineering - PPG-CEM ,
(d)
Professor, Department of Natural Sciences, GPQM - Research Group of Materials Chemistry,
Federal University of São João del Rei - UFSJ, São João del Rei, MG, Brazil.
(e)
Professor, Laboratory of Solid State Chemistry, State University of Campinas (UNICAMP),
Campinas, Brazil.
Address for correspondence: Andréa Cândido dos Reis, Department of Dental Materials and
Prosthesis – School of Dentistry of Ribeirão Preto - FORP-USP, Av. do Café, s/nº, CEP: 14040-
Abstract
Objective: The purpose of this study was evaluate, for the first time, the impact of incorporation of
Design: The β-AgVO3 was synthesized and characterized by X-ray diffraction (XRD), Fourier
(SEM/EDS). Resins specimens were prepared with 0-10% wt.% β-AgVO3 and characterized by
SEM, XRD and optical microscopy. The antibiofim activity of the samples against Candida
albicans and Streptococcus mutans was investigated by XTT reduction test, colony-forming units
(CFUs), and confocal laser scanning microscopy (CLSM). The flexural strength, hardness, and
surface roughness of the samples containing β-AgVO3 were compared with the pure PMMA
matrix.
Results: The incorporation of 10% β-AgVO3 significantly reduced the metabolic activity of C.
albicans and S. mutans (p<0.05). There was a reduction in microbial load (CFU/mL) of
microorganisms for the different concentrations used (p<0.05), which was confirmed by confocal
microscopy. The addition of β-AgVO3 did not change the mechanical properties of hardness and
surface roughness of the resins (p>0.05). However, flexural strength decreased with the addition
of main microorganisms associated with dental prostheses. However, the viability of clinical use
behavior.
Introduction
prostheses plays an important role in the development of caries, periodontal disease and mucositis
(Rupf et al., 2012; Queiroz et al., 2013). These factors limit the longevity of rehabilitative treatment
and constitute a risk of opportunistic infections, reducing quality of life and generating additional
costs to patients (Larazin et al., 2013). The colonization process on the surface of dental prostheses
is characterized by various steps (Rupf et al., 2012) and occurs because acrylic resins have porosity,
an absence of ionic charge on the surface, roughness, and a capacity to absorb fluids, all of which
lead to the accumulation of microorganisms (Santos, Pithon, Carvalho, Ramos & Romanos, 2013;
Sivakumar et al., 2014). The development of dental acrylic resins capable of inhibiting biofilm
formation is therefore critical in the control of oral disease (Wang, Shen & Haapasalo, 2014).
In an effort to add antimicrobial activity to these materials, studies have used additives such
aluminum borate (Redding et al., 2009; Alcântara et al., 2012; Han et al., 2015). The antimicrobial
properties of silver (Ag) dates to 3000 years ago, and the mechanism is based on the interaction of
silver with thiol groups of enzymes involved in bacterial cell metabolism thus causing cell death
(Kwakye-Awuah, Williams, Kenward & Radecka, 2008). Due to this property, silver ions and
silver nanoparticles (AgNPs) have also been introduced in dental materials (Bürgers et al., 2009;
Yang, Zhang, Yu, Zhen & Huang, 2014). For AgNPs these properties appear to be even better in
function to their nanoscale dimensions, interact extensively with microorganisms (Fan et al., 2011;
Holtz, Lima, Souza Filho, Brocchi & Alves, 2012; Kumar, Jolivalt, Pulpytel, Jafari & Arefi-
Khonsari, 2013; Hojati et al., 2013). However, in addition to great aesthetic inconvenience (color),
there is the difficulty of stabilizing nanoparticles when used alone (Holtz, Lima, Souza Filho,
Brocchi & Alves, 2012). AgNPs tend to cluster, which may negatively influence in their
antimicrobial effect by reducing the surface contact area (Shameli et al., 2011).
A possible alternative to overcome this problem was recently proposed and is to support
AgNPs on Ag nanowires vanadate (β-AgVO3) (Holtz et al., 2010). This nanomaterial has
antimicrobial property as it the silver and the vanadium elements act synergistically and interact
with microorganisms' cell membranes. In addition, this compound has a high dispersion of silver
nanoparticles on silver vanadate nanowires, maintaining high surface contact with the
microorganisms (Holtz et al., 2010; Holtz, Lima, Souza Filho, Brocchi & Alves, 2012). Previous
studies have shown the possibility of using this hybrid nanomaterial as an antimicrobial additive
for water-based paint for domestic or hospital environment in order to improve sanitary conditions
and reduce the number of hospital infections once it exhibits antibacterial activity 30 times larger
than that of Oxacillin (Holtz et al., 2010) and as an additive for acrylic resin for using in dental
applications, as it functionalized samples with this nanomaterial showed inhibition zone against
the growth of the main microorganisms in the oral cavity demonstrating the need for studies on
biofilm models as well as the mechanical properties (de Castro et al., 2014).
The objective of this study was therefore to evaluate the antibiofilm activity of two acrylic
resins containing β-AgVO3 against C. albicans and S. mutans, the main microorganisms associated
with dental prostheses. The hardness, surface roughness, and flexural strength of the resins were
also evaluated, as these mechanical properties, together with antibiofilm action, are factors that are
reaction between silver nitrate (AgNO3, Merck 99.8%) and ammonium metavanadate (NH4VO3,
Merck 99%) according to the methodology described by Holtz et al (2010). Initially, 1.3569 g of
AgNO3 and 0.9736 g of NH4VO3 were each solubilized in 200 mL of distilled water. The solutions
were stirred separately on a 65 °C heated surface for 10 min. Next, the silver nitrate solution was
added dropwise using a burette into the ammonium metavanadate solution under constant stirring
at 65 °C. The precipitate obtained was washed with distilled water and absolute ethanol several
Characterization of β-AgVO3
The silver vanadate was characterized to ensure standardization of the resulting materials.
For this purpose, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR),
scanning electron microscopy, and microanalysis (SEM/EDS) were used. XRD analysis was
radiation (λ = 1.5406 Å) at 40 kV and 30 mA. Data were collected continuously from 10° < 2θ <
70° with a velocity of 1°/min to identify the crystal structure. The crystallinity index (IC) of the
sample was determined (Krimm & Tobolsky, 1951). The IC (%) was calculated from the
ratio between the crystalline diffracted area (AC) and the total area of diffraction (AT =
(1)
Infrared spectra with Fourier transform were collected on an FTLA 2000 – 102
spectrometer using transmittance mode in the region of 4000-400 cm-1 to monitor β-AgVO3
synthesis. Finally, the morphological analysis of β-AgVO3 was performed by scanning electron
microscopy (SEM) on a Magellan 400L FEI microscope coupled with an energy dispersive X-ray
thermopolymerizable (TR) (Clássico Artigos Odontológicos®) acrylic resins were used. Six
experimental groups functionalized with β-AgVO3 at concentrations of 0.5, 1, 2.5, 5 and 10%
(wt.%) and a control group without additive were formed. The β-AgVO3 was added to the polymer
particle, and after homogenization, the monomer was added at a ratio of three parts powder
(polymer) to one part liquid (monomer) according to the manufacturer's recommendations. During
the plastic phase, the resin was placed in holes prepared in metal muffles with appropriate
dimensions for each test and was cured according to the manufacturer's recommendations. For the
antibiofilm tests, surface roughness was standardized at 0.2 µm using a Surftest SJ-201P (Mitutoyo
Corporation).
Characterization of functionalized resins
The characterization of the samples with β-AgVO3 was performed in comparison with
control samples without nanomaterial by XRD and SEM order to identify the changes introduced
by the incorporation and the pattern of dispersion of the particles. These techniques were performed
as described previously for the characterization of β-AgVO3. The morphology of resins was also
analyzed by optical microscopy using a Leica MZ7.5 stereomicroscope coupled with a DFC-
490 camera.
Antibiofilm experiment
Two standard strains were used in the study: Candida albicans (ATCC 10231) and
Streptococcus mutans (ATCC 25175). For inoculum preparation, the microorganisms were seeded
in Petri dishes with specific culture media (Sabouraud Dextrose Agar for yeast and modified SB-
20 Agar for bacteria). After incubation at 37°C for 48 h, standardized S. mutans (108 CFU / mL)
and C. albicans (106 CFU / mL) microbial inocula were obtained in 0.85% saline using a PCB 687
spectrophotometer (BYK Gardner). The biofilm was formed in 24-well polystyrene plates. Sterile
resin samples were placed individually in each well, and 1000 µL of the inoculated culture medium
was transferred to each hole. The plates were incubated at 37°C for 1 h 30 min with stirring at 750
rpm in a bacteriological incubator (Incubator Shaker Mod. - EC-320, Cienlab) for adherence of the
biofilm. After the adhesion period, the specimens were rinsed with 2000 µL of 0.85% saline to
remove non-adherent cells, to buffer the medium, and to remove metabolites. Next, 1000 µL of
sterile culture medium was added to each well to provide more nutrients for the microorganisms.
The plates were incubated at 37°C with 750 rpm stirring for 48 h for biofilm maturation.
Biofilm viability was evaluated using the XTT reduction test ([2,3-bis (2-methoxy-4-nitro-
formation, each specimen was washed once with saline, inserted into a polypropylene tube with
3000 µL of PBS, and sonicated (40 Hz) for 20 min to detach the biofilm. Then, 100 µL aliquots of
homogenized tube suspension and 50 µL of XTT solution were transferred to each well of a 96-
well plate. The plates were incubated in the dark at 37 ° C for 24 h to measure the formation of
Instruments). The number of viable cells was quantified in terms of colony forming units per
milliliter (CFU/mL) after sowing 25 µL of the raw and diluted aliquots (n = 8) in specific culture
media and incubating at 37 °C for 24 h. Qualitative analyses of the biofilms (48 h) were performed
for the control group and for samples containing 2.5, 5 and 10% β-AgVO3. The specimens were
stained using a Live Dead/Back Light Kit (Molecular Probes, Eugene) and incubated for 15 min
under light at room temperature. An image of each group was captured with a confocal laser
Mechanical tests
For mechanical testing, n = 10 was used for each group. The three-point flexural strength
test was conducted on an EMIC DL 2000 universal testing machine at a speed of 5 mm/min and a
load of 20 kgf according to the ISO 20795-1:2008 standard, using specimens with dimensions 65
mm x 10 mm x 3.3 mm. The analysis of surface roughness was carried out according to technical
standard NBR ISO 4287:2002 using Surftest SJ-201P (Mitutoyo Corporation); the analyzer tip
touched the piece and covered a distance of 4 mm, making three measurements on each specimen
hardness tester. Three randomly equidistant measurements were performed on each specimen using
Statistical analysis
the concentration of β-AgVO3 (on 6 levels: 0%, 0.5%, 1%, 2.5%, 5%, and 10%) and acrylic
resin (on 2 levels: SC and TR). After the normal (Shapiro-Wilk test) and homogeneous
(Levene test) distribution of data was verified, 2-way ANOVA, followed by the Tukey test
(α= 5%) was used. Statistical analysis was performed with software (SPSS v17.0; SPSS Inc).
Results
crystallographic card JCPDS 29-1154. The crystallinity index of the sample was estimated to
be 67% according to the background displayed by XRD (Krimm & Tobolsky, 1951). This
result indicates that the sample is not completely crystalline and contains 33% amorphous phase.
The diffraction peaks located at 2θ of 37.79 and 44.10 correspond to the presence of nanoparticles
of metallic silver, Ag0 (JCPDS 87-597), which formed over the entire surface of the crystals (Figure
1).
The main bands observed are located between 400-1000 cm-1, most of them attributed to
bonds between the oxygen and vanadium atoms. In the spectrum, the band located at 1633 cm-1
corresponds to the vibration of the −OH bond of the water molecule (Figure 2).
The strong band observed at 1409 cm-1 in ammonium vanadate is attributed to N−H
symmetric stretching. This band completely disappears in the β-AgVO3 spectrum, indicating its
formation, which is also illustrated by the intensification of the band at approximately 849 cm-1
associated both with the vibration of V−O stretching and with the vibration mode of the Ag−O−V
bond. The bands located at 965, 925, 896 and 653 cm-1 in the spectrum of β-AgVO3 are attributed
to V=O stretching, while the band located at 501 cm-1 may be associated with the V−O vibration
mode. The bands in the region 940-775 cm-1 also suggest the presence of polymeric vanadate
groups. As with water, the presence of silver in the system also affects the vibration mode of the
metal-oxygen bonds, contributing to the displacement of certain bands in relation to the ammonium
vanadate spectrum. This effect could be observed especially for the bands in the 700 cm -1 region.
It can be observed that the β-AgVO3 showed acicular crystals, with a mean diameter of 150
nm and length in the order of micrometers. The EDS spectrum reveals the presence of the elements
silver, vanadium and oxygen, which is consistent with the established formulation for the synthesis
of the material. The peak located at 0.28 keV is attributed to elemental carbon (C), used in tape
Comparing the resins with different β-AgVO3 content, it appears that regardless of
the content and type of resin, there are always β-AgVO3 domains throughout the matrix,
following a standard circular dispersion (Figure 4). The dispersion mode presented by β-
AgVO3 can be related to the particle size presented by the polymer powder used in the
preparation of the resins. The β-AgVO3 crystals appears to have been confined to the area
10% β-AgVO3. Is observed the presence of clusters, which indicates that the method used to
prepare the resins was unable to promote good dispersion of β-AgVO3 in the polymeric
matrix.
X-ray diffraction of the resins revealed that the incorporation of β-AgVO3 in percentages
above 1% resulted in segregation in the system and that it is possible to identify the vanadate
diffraction peaks in the polymeric matrix. This phenomenon becomes more evident when the β-
The XTT reduction and count of colony-forming units per milliliter (CFU/mL) analyses
showed that the metabolic activity of C. albicans and Streptococcus mutans biofilms decreased
significantly as the β-AgVO3 content in the resins increased. The absorbance values obtained using
XTT on C. albicans were reduced significantly compared to that of the control group for both resins
(p = 0.368) when concentrations of 5 and 10% were used (p < 0.05). However, for S. mutans, a
greater reduction was observed only with the addition of 10% (p = 0.023) (Figure 7).
With the incorporation of 10% β-AgVO3, the number CFUs of C. albicans was reduced
compared to that of the control group (p = 0.000) for both resins (p = 0.431). For S. mutans, this
reduction was observed with the incorporation of 2.5% followed by 5 to 10% (Figure 8).
The images obtained by confocal laser scanning microscopy (CLSM) of the metabolic
activity and amount of C. albicans and S. mutans biofilm formed on the surface of certain TR
specimens. Greater cell density and a higher proportion of green cells were observed in the control
group than in the other groups for both micro-organisms. A change in cell viability (increased ratio
samples containing 10% additive, almost complete inhibition was observed (Figure 9).
Mechanical properties
For SC, the incorporation of 0.5% β-AgVO3 resulted in a significant increase in surface
hardness values (26.14 ± 5.13) compared to those of the other groups (p=0.000). The surface
hardness of the TR remained unchanged after the incorporation of different fractions of β-AgVO3
(p=1.000). When comparing the two kind of resins, a significant difference could be observed only
The flexural strength of both resins was reduced when a 2.5% concentration was
control group (p=0.000), which, in turn, showed similar results to the groups containing
There was no statistically significant difference in the surface roughness of the acrylic resins
(p=0.209) when the samples containing β-AgVO3 were compared to the control group (p=0.751).
This result indicates that the polishing process of functionalized resin samples provides a surface
similar to conventional resin and does not promote an uneven surface that would encourage the
This study demonstrated that nanostructured silver vanadate (β-AgVO3) can be easily
synthesized by the precipitation reaction between silver nitrate and ammonium metavanadate. The
β-AgVO3 formed had acicular morphology with a mean crystal diameter of 150 nm and length in
the order of micrometers, similar to the pattern observed in the images obtained by
transmission electron microscopy (STEM) by Castro et al (2015). The sample was not fully
Upon completion of the synthesis and confirmation of the formation of β-AgVO3 by X-ray
diffraction and FTIR spectrum, different amounts of β-AgVO3 were incorporated into two acrylic
resins used in the manufacture of dental prostheses. The antibiofilm action and mechanical
properties of the resulting materials were then evaluated, as these factors are directly related to the
in dentistry practice (Larazin et al., 2013; Sato, Ohshima, Maeda & Ohkubo, 2013), this study used
CFU, XTT reduction, and CLSM to evaluate the modified resins' antibiofilm activity. As in studies
involving other compounds, the antimicrobial effect was dose dependent (Fan et al., 2011; Shameli
et al., 2011; de Castro et al., 2014; Han et al., 2015). In the analysis of cellular metabolism using
the XTT method, a decline in the metabolism of C. albicans was observed with the incorporation
of 5 and 10% β-AgVO3. For S. mutans, this decline was clearly evident only at a concentration of
10%. The CFU count showed a significant reduction in the number of viable microorganisms when
vanadate was incorporated at percentages of 2.5, 5 and 10% for S. mutans and 10% for C albicans.
Note that there was no complete correlation between the methodologies used in this research
because of the difference in sensitivity of each technique as well as the morphological and
metabolic characteristics of each microorganism. However, in general, the higher the concentration
of β-AgVO3 incorporated into the resins, the higher the antibiofilm activity observed. CLSM
analysis qualitatively complemented the previous analyses, showing a reduction in the amount of
prerequisite for the colonization of dental prostheses which will function as a reservoir of
infection. Thus, the resin surface topography is critical for the biofilm accumulation,
interfering with the success of rehabilitation treatment and quality of life of patients
In dental literature, the surface roughness values of acrylic resins can range from
0.02 to 7.6 µm. Smooth and highly polished surfaces are important not only for the patient's
comfort, but also for denture longevity, good aesthetic results and base plate retention
(Akalın-Evren, Kulak-Özkan, Ozcan & Kadir, 2014). The presence of antimicrobial agents
may be useless if the tested acrylic resin is able to retain greater amounts of debris. The
values of surface roughness in this study were close to 0.2 µm, regardless of the concentration
of the nanomaterial incorporated in resins. This value which can be considered a value below
which microbial adhesion is not expected (Bollen, Lambrechts & Quirynen, 1997). These
results demonstrate that the incorporation of β-AgVO3 did not impossible to obtain a smooth
In order to predict the performance of the materials when exposed to the forces of the oral
cavity, hardness and flexural strength tests performed. Hardness in the case of dental resins, may
be associated with forces arising from occlusion and mechanical cleaning as well as contact with
oral fluids that can alter the surface properties of the prosthesis (Ayaz & Durkan, 2013; Goiato et
al., 2013). The results showed that the surface hardness of SC increased significantly with the
incorporation of 0.5% β-AgVO3, suggesting that vanadate had no negative effect on the degree of
resin polymerization. This phenomenon is observed because, in general, hardness shows a positive
correlation with the degree of polymer conversion (Faltermeier, Rosentritt & Müssig, 2007). At
concentrations above 0.5% β-AgVO3 and for TR, hardness remained practically unchanged.
Finally, flexural strength is an important property for dental resins, as the major cause of
prosthesis fracture in the oral cavity is related to stresses caused by repetitive chewing movements
(Ayaz & Durkan, 2013). According to ISO 20795-1:2008 (ISO, 2008), the flexural strength of
polymeric materials must be at least 50 MPa. Based on the flexural strength values obtained,
to the control group. The incorporation of 10% promoted results below those required by this
standard, the average being 48.03 MPa. Castro et al (2015) found that incorporation of 5 and
10% β-AgVO3 causes a reduction in impact strength of acrylic resins which may increase the
Generally, composite materials with lower mechanical properties that of the matrix are
obtained when the filler content is below the critical concentration or when it forms clusters that
weaken the system. It is believed that for the system in question, the non-improvement in resistance
with the addition of low concentrations of β-AgVO3 and the significant reduction observed with
concentrations of 5 to 10% was caused by the dispersion mode and the formation of agglomerates.
Scanning electron micrographs revealed that the β-AgVO3 dispersion along the polymeric
matrix follows a circular mode, with an increase in the number of domains in the resins with higher
β-AgVO3 content. This distribution can be explained by the fact that when β-AgVO3 is added to
the mixture to prepare the resin, it disperses in the monomer, being confined mainly around the
large polymer particles to the extent that polymerization of the system occurs. As the polymer
particles already existed, vanadate dispersion of the resin was limited. It should also be noted that
increasing the β-AgVO3 fraction from 2.5 to 5 and 10% caused the formation of clusters in the
system, a result that could be enhanced by X-ray diffraction of the resins and optical microscopy.
As the β-AgVO3 was confined to the area surrounding preexisting polymer particles during
the curing stage, there was no possibility of better dispersion, and the increase of its fraction in the
system caused the formation of agglomerates. These agglomerates can act as a stress center and
adversely affect the mechanical properties of resins, especially properties involving more complex
forces, such as bending resistance (Han, Kiat-Amnuay, Powers & Zhao, 2008; Sodagar et al.,
2012). These results are consistent with the findings of the studies of Sodagar et al (2012) and
Shibata et al (2007), when adding silver nanoparticles to acrylic resins, reported that they can act
as impurities in the internal structure of the polymerized material, weakening the system.
The attempt to functionalizing the resin with β-AgVO3 through a simplistic method of
manual manipulation of easy access to the dentist in clinical practice and low processing costs,
proposal in the present study was unable to promote effective dispersion of inorganic
particles in polymeric matrix. The reduced chemical affinity between the inorganic filler of
hydrophilic nature and the polymer, predominantly hydrophobic, may be responsible for the
spatial confinement of the polymer chains. As an approach to overcome this problem other
methods of synthesis and the incorporation should be proposed, among which attempting to
improve the β-AgVO3 compatibility with the polymeric matrix by means of chemical surface
polymer solution followed by solvent removal (Utracki, 2008), use of mixers such as amalgam
mixer (Hamedi-Rad, Ghaffari, Rezaii & Ramazani, 2014) or extruder (Soares et al., 2015)
Considering the method proposed in this study, it may be concluded that acrylic resins
functionalized with β-AgVO3 antibiofilm exhibit significant activity against C. albicans and S.
mutans. The development of new methodologies to improve the dispersion pattern of β-AgVO3 in
a polymer matrix may provide the materials with better mechanical properties and further expand
their antibiofilm action by exposing a larger surface area. Furthermore, analyses that evaluate the
possible cytotoxicity of these materials are required to ascertain the viability of their use.
Conclusions
In this work we have successfully demonstrated that the addition of different concentrations
of β-AgVO3 to dental acrylic resins can promote antibiofilm activity against C. albicans and S.
mutans. This makes the material promising to use for fabrication of dental devices. The dispersion
of β-AgVO3 in the resins suggests that the mixing procedure adopted in this work was not efficient
enough to disperse the resins evenly in the system. Therefore, new methodologies are currently
Conflict of interest
The authors declare that they have no conflict/s of interest related to the present study.
Acknowledgements
We thank the São Paulo Research Foundation – FAPESP (process numbers: 2012/09124-
1 and 2012/09347-0) for funding the study and the Solid State Chemistry Laboratory - UNICAMP
and Oral Rehabilitation Research Laboratory FORP / USP for the facilities.
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Fig. 4 Scanning electron micrograph of the polymer used in the preparation of the resin (a)
Fig. 5 Optical microscopy of SC acrylic resin containing 5% β-AgVO3 (a) and TR acrylic
Fig. 7 Mean absorbance of XTT and Standard Deviation: Identical letters indicate statistical
equality for C. albicans, while equal numbers indicate equality for S. mutans.
Fig. 8 Mean number of colony forming units and Standard Deviation: Identical letters indicate
statistical equality for C. albicans, while equal numbers indicate equality for S. mutans.
Fig. 9 Confocal laser microscopy of the formed biofilm: Candida albicans (a) TR 0%; (b) TR 5%;
(c) TR 10%; Streptococcus mutans (d) TR 0%; (e) TR 5%; (f) TR 10%.
Fig. 10 Flexural strength (MPa) and Standard Deviation: Identical letters indicate statistical equality
Fig. 11 Surface roughness (µm) and Standard Deviation: Identical letters indicate statistical
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Table 1. Mean (SD) of the surface hardness Knoop (HK) of resins containing β-AgVO3
* SC – Autopolymerizable ; TR - Thermopolymerizable
* Lowercase letters - comparison between lines
* Uppercase letters - comparison between columns
* Equal letters - statistical equality