UNIVERSIDADE ESTADUAL PAULISTA

“JÚLIO DE MESQUITA FILHO”

Instituto de Ciência e Tecnologia

Campus de São José dos Campos

ORIGINAL ARTICLE DOI: https://doi.org/10.4322/bds.2023.e3672

Braz Dent Sci 2023 Jan/Mar;26 (1): e3672

Antimicrobial eﬃcacy of S-PRG containing toothpastes on

S. mutans

bioﬁlm development

Eﬁcácia antimicrobiana de dentifrícios contendo S-PRG sobre o desenvolvimento de bioﬁlme de

S. mutans

Manuela da Silva SPINOLA1 , Jacqueline Landi MENDONÇA2 , Maíra Terra GARCIA3 , Taciana Marco Ferraz CANEPPELE2 ,

Juliana Campos JUNQUEIRA3 , Carlos Rocha Gomes TORRES2 , Alessandra Buhler BORGES2 

1 - Faculdade de Odontologia da Universidade Braz Cubas, Mogi das Cruzes, Brazil.

2 - Instituto de Ciência e Tecnologia da UNESP, Departamento de Odontologia Restauradora, São José dos Campos, Brazil.

3 - Instituto de Ciência e Tecnologia da UNESP, Departamento de Biociências e Diagnóstico Oral, São José dos Campos, Brazil.

How to cite: Spinola MS, Mendonça JL, Garcia MT, Caneppele TMF, Junqueira JC, Torres CRG, et al. Antimicrobial efcacy of S-PRG

containing toothpastes on

S. mutans

biolm development. Braz Dent Sci. 2023;26(1):e3672. https://doi.org/10.4322/bds.2023.e3672

ABSTRACT

Objective: to investigate the antimicrobial effects of toothpastes containing bioactive surface pre-reacted glass

particles (S-PRG) on

S. mutans

biolms adherence, initial colonization and maturation. Material and Methods:

a reference UA 159 and a clinical

S. mutans

(SM6) strain were used. Bovine enamel specimens were randomly

allocated into the groups (n=5): toothpastes containing 0%; 1%; 5%; 20%; 30% S-PRG; positive control

dentifrice (NaF+triclosan); and negative control (distilled water). For biolm development, samples were

placed in a 24-well plate containing articial saliva (4h), followed by adding 1mL of articial saliva, BHI broth

and 225μL of

S. mutans

suspension. Treatments with toothpastes were applied previously or after 4h and 24h

of biolm formation. Samples were incubated for 48h at 37°C in 5%CO2 and biolm was detached and seeded

in Petri dishes for determining the number of viable cells. Data were analyzed by ANOVA and Tukey test (5%).

Results: signicantly lower microorganisms’ adherence (p<0.05) was obtained for all S-PRG toothpastes, with

similar results to NaF+triclosan for SM6 and 20 and 30%S-PRG groups exhibiting higher inhibition effect than

the NaF+Triclosan for UA159. Antibacterial effect on the early-stage biolm was also observed for the S-PRG

groups, but was not superior to the NaF+Triclosan toothpaste. For the mature biolm, the effective antimicrobial

potential of S-PRG toothpastes was observed only for the SM6 clinical strain, but was not higher than the positive

control. Conclusion: experimental S-PRG toothpastes were effective to inhibit

S. mutans

biolm growth by

exhibiting antimicrobial activity, being promising agents to prevent cariogenic biolm development.

KEYWORDS

Biolm; Dental enamel; Giomer;

Streptococcus mutans

, S-PRG.

RESUMO

Objetivo: investigar o efeito de dentifrícios contendo S-PRG sobre a colonização inicial e maturação de biolmes de

S. mutans

. Material e Métodos: uma cepa de referência (UA 159) e uma cepa clínica de

S. mutans

(SM6) foram

utilizadas. Espécimes de esmalte bovino foram alocados nos grupos (n=5): dentifrícios contendo 0%; 1%; 5%;

20% e 30%S-PRG; controle positivo (NaF+triclosan); e controle negativo (água destilada). Os espécimes foram

inseridos em uma placa de 24 poços contendo saliva articial (4h), seguido por adição de 1mL de saliva articial,

BHI, 225μL de suspensão de

S. mutans

e foram tratados com suspensões de dentifrícios antes ou depois de 4 e

24h da formação do biolme. Os espécimes foram incubados por 48h e o biolme foi removido dos espécimes e

semeado em placas de Petri para contagem de UFC/mL. Os dados foram analisados por ANOVA e teste de Tukey

(5%). Resultados: houve diminuição na adesão de microrganismos para os grupos tratados com S-PRG (p<0.05).

Para SM6, os dentifrícios contendo S-PRG apresentaram resultados semelhantes ao NaF+triclosan e para a cepa

Braz Dent Sci 2023 Jan/Mar;26 (1): e3672

Spinola MS et al.

Antimicrobial efficacy of S-PRG containing toothpastes on S. mutans biofilm dev elopment

Spinola MS et al. Antimicrobial eﬃcacy of S-PRG containing toothpastes on

S. mutans bioﬁlm development

INTRODUCTION

Dental caries is a biolm and sugar-dependent

disease that results in mineral loss of tooth surface.

The control of the disease is directly related to

the biolm monitoring, which can be achieved

by means of a controlled diet, with less intakes of

carbohydrates, and with efcient oral hygiene, to

disaggregate the cariogenic biolm formed over

the tooth surface [1,2]. Although these methods

are well established, and the prevalence and

severity of dental caries have declined in recent

years, the cariogenic biolm tends to remain in

regions that are difcult to access by brushing, such

as the occlusal pit and ssures, and interproximal

regions, making it still one of the most prevalent

diseases at a world level [3].

Strategies to improve the control of the

acidogenic microorganisms that are prevalent

in the cariogenic biofilm using products with

multiple biological effects such as antibacterial

properties and biolm formation inhibition have

been tested [4,5].

The surface pre-reacted glass ionomer

(S-PRG) filler is an innovative bioactive

material composed by a 3-layer structure

derived from a proprietary technology.

A fluoroboroaluminosilicate multifunctional

glass is covered with polysiloxane, creating a

porous superficial silica-glassy layer. Then, a

polyacrylic acid solution is sprayed on the particle

and penetrates the surface porosities, reacting

with the inner glass core. This creates a thin

intermediate stable pre-reacted glass ionomer

phase [6]. The dental products containing S-PRG

llers are named GIOMER, due to their “Glass

IOnoMER” phase. The bioactive effect of these

materials is exerted by the multi-ions release

(uoride, strontium, borate, sodium, aluminum,

and silicate) derived from the multifunctional

glass in the presence of moisture [7].

These fillers can be incorporated

into preventive and restorative materials.

The antimicrobial/anticaries/remineralizing/

desensitizing effects have been shown

with surface barrier materials, varnishes,

and sealants [8-11]. Favorable results have

been observed regarding the antimicrobial

activity of restorative materials using Giomer

technology [12,13], besides the ability to

reduce the demineralization of dentin adjacent

to restorations [14], and exert acid-buffering

properties, which might reduce the susceptibility

to secondary caries [15].

This motivated the development of

toothpastes containing the S-PRG particles,

providing the benet of releasing the multi-ions

into the oral environment, as an alternative

for caries prevention. Studies have shown

that experimental S-PRG based toothpastes

were effective on the enamel demineralization

prevention [16-18]. However, despite previous

studies present promising results regarding the

addition of S-PRG to dental products, little is

still known about the ideal concentration of

these particles in dentifrices concerning their

antimicrobial efficacy on dental cariogenic

biofilms, aiming to promote strong scientific

evidence so that commercial products can be

developed and be further available in the market.

Thus, the present study reports the ndings on

the inhibitory effect of experimental dentifrices

containing different concentrations of S-PRG on

adherence, initial colonization and maturation

phases of

S. mutans

biolms. The null hypothesis

tested was that there is no difference in the

mutans

biolm inhibition among the toothpastes

in the different phases tested.

MATERIAL AND METHODS

Ethical aspects

The study protocol was approved

by the local Ethics Committee in Research

(CAAE:005611/2017).

UA159 o dentifrício com 30%S-PRG apresentou efeito superior. Efeito antimicrobiano no biolme recém-formado

(4h) foi observado para os grupos contendo S-PRG, mas não foi observado efeito superior ao NaF+Triclosan. Para

o biolme maduro, efeito antimicrobiano do S-PRG foi observado apenas para a cepa clínica, mas não superior ao

efeito do NaF+Triclosan. Conclusão: dentifrícios contendo S-PRG foram ecazes na inibição do desenvolvimento

de biolmes de

S. mutans,

sendo promissores agentes para prevenir o desenvolvimento de biolme cariogênico.

PALAVRAS-CHAVE

Biolme; Esmalte dental; Giomer;

Streptococcus mutans

; S-PRG.

Braz Dent Sci 2023 Jan/Mar;26 (1): e3672

Spinola MS et al.

Antimicrobial efficacy of S-PRG containing toothpastes on S. mutans biofilm dev elopment

Spinola MS et al. Antimicrobial eﬃcacy of S-PRG containing toothpastes on

S. mutans bioﬁlm development

Microorganisms

In this study, one reference

S. mutans

strain (UA159) and one clinical strain were

used. The clinical strain was previously isolated

from the active cavities of a subject according

to Carvalho et al. (2006) [19], was identied

by PCR [20], and confirmed by automatic

sequencing [21], named SM6.

Study design

Six independent experiments were conducted

(three for the

S. mutans

UA159 strain, and three for

the SM6 strain). Each experiment presented one

factor under investigation (treatment toothpastes)

at 7 levels (distilled water as negative control (DW);

commercial NaF+Triclosan-based toothpaste

as positive control; and toothpastes containing

S-PRG: 0%; 1%, 5%, 20%, and 30%). For both

strains, the following effects were investigated:

the inhibitory effect on biolm adherence, initial

colonization and maturation (n = 5/group).

UFC/mL was the response variable.

Specimens’ preparation

Fresh, non-damaged bovine incisors were

collected for this study. The crowns were

separated from the roots and stored into 0.1%

thymol solution at 4ºC until required. Seventy

cylindrical enamel specimens (6 mm in diameter)

were obtained from the labial surface of the teeth,

using a custom-made diamond-coated trephine

mill adapted to a circular cutting machine.

The enamel specimens were polished using SiC

sandpapers in sequential grits of 1200, 2400, and

4000 (FEPA-P, Struers, Copenhagen, Denmark),

under constant water irrigation for 10, 20 and

30 s, respectively, using an automatic polishing

machine (Tegramin 25, Struers). After each paper

grit change, specimens were kept in an ultrasonic

bath for 5 min, to remove debris and abrasive

grains. The specimens were examined under a

stereomicroscope (Carl Zeiss – Stemi 2000 - 20X)

to ensure the absence of cracks or other surface

defects [22]. The specimens were autoclaved

before the beginning of the experiments.

Preparation of the standardized suspension

S. mutans

The microorganisms were cultured in brain-

heart infusion broth (BHI Himedia, Mumbai,

India) at 37oC for 48 h (5% CO2). The microbial

cells in the culture were centrifuged (1300 rpm

for 10 min), and the pellet was rinsed twice

with 0.85% NaCl (Labimpex, Sao Paulo, Brazil).

The cell suspensions were adjusted to 108 cells/mL

using a spectrophotometer at a wavelength of

398 nm and optical density of 0.620 (B5B2,

Micronal, São Paulo, Brazil) [23].

Groups division

For each experiment, testing both the

reference and the clinical strain, the specimens

were divided in seven groups (n = 5 each).

The experimental toothpastes were prepared by

Shofu Inc. (Kyoto, Japan), and contained silicic

anhydride, sodium carboxymethylcellulose,

glycerol, sorbitol solution, perfume, and 1 μm

S-PRG particles: 0% (placebo), 1%, 5%, 20%,

and 30% (in weight). The distilled water was the

negative control and a commercial toothpaste

containing NaF (1450 ppm F-), Triclosan,

PVM/MA copolymer, sodium lauryl sulfate,

sorbitol, carrageenan, aroma, and hydrated silica

(Colgate Total 12, Colgate-Palmolive, São Paulo,

Brazil) was the positive control.

S. mutans

biolm formation

Specimens were placed in 24-well plates

containing 2 mL of articial saliva containing

0.33g KH2PO4; 0.34g Na2HPO4; 1.27g KCl; 0.16g

NaSCN; 0.58g NaCl; 0.17g CaCl, 0.16g NH4Cl;

0.2g urea; 0.03g glucose; 0.002g ascorbic acid

and 2.7g mucin [24] and incubated for 4 h at

37oC to allow the formation of a surface pellicle.

After 4h, specimens were rinsed twice with 2 mL

of PBS solution with the use of a pipette and

then 1 mL of BHI broth supplemented with 5%

sucrose and 1 mL artificial saliva were added

to each well. Then, 225 μL of the standardized

S. mutans

suspension was added to each well,

and the plates were incubated for 24 h at 37°C

(5% CO2). The wells were washed three times

with 2 mL PBS for the removal of weakly adhered

cells with the use of a pipette, and 1 mL of fresh

BHI broth supplemented with 5% sucrose and

1 mL of fresh articial saliva was again added

to each well. The plates were incubated for an

additional 24 h at 37oC.

Antibacterial potential of the toothpastes on

S. mutans

biolm development

Specimens were treated with dentifrices

slurries prepared in artificial saliva (1:3) for

Braz Dent Sci 2023 Jan/Mar;26 (1): e3672

Spinola MS et al.

Antimicrobial efficacy of S-PRG containing toothpastes on S. mutans biofilm dev elopment

Spinola MS et al. Antimicrobial eﬃcacy of S-PRG containing toothpastes on

S. mutans bioﬁlm development

5 min in 3 different phases: before the immersion

in articial saliva to evaluate the effect of the

treatments on the microorganisms adherence

phase, after 4 h of biolm growth to evaluate

the antibacterial potential over the early-stage

biofilm, and after 24 h of biofilm growth to

evaluate the antibacterial potential over biolm

maturation.

Cell count for biolm analysis

After nal incubation, all specimens (treated

with the slurries before, after 4h and 24h of

biolm growth) were transferred to Falcon tubes

containing 4 mL PBS. The biofilm was then

detached from the specimens using an ultrasonic

homogenizer (Sonoplus KD2200, Bandelin

Eletronic, Berlin, Germany) at 7 W for 30 s.

Serial dilutions were prepared from the obtained

solution and plated on Petri dishes containing

BHI agar. The plates were incubated for 48 h

at 37oC (5% CO2) to determine the number of

colony-forming units (CFU/mL) [23].

Statistical analysis

Data of CFU/mL were transformed in

log10 and means ± standard deviation were

calculated for each treatment. The Shapiro-Wilk

and Brown-Forsythe tests were used to assess

the normality and homoscedasticity of the data.

They were then analysed by one-way ANOVA

followed by Tukey’s test with a p-value set at 5%,

using Statistica for Windows software (StatSoft,

Hamburg, Germany).

RESULTS

Potential of the toothpastes on

S. mutans

adherence

Figure 1a-1b show mean and standard

deviation data for the tested groups, using the

reference UA159 and the SM6 clinical strains,

respectively. The results of ANOVA one-way

indicated signicant difference for the groups

(p<0.05). The inhibiting effect was higher

when more concentrated toothpastes were used,

indicating a dose-dependent effect. According to

Tukey’s test, the toothpastes containing S-PRG

were capable to inhibit

S. mutans

adherence

when compared to the negative control (p<0.05)

for both strains. For UA159 strain, both 20 and

30% S-PRG toothpastes showed significantly

lower

S. mutans

adherence than the positive

commercial toothpaste containing NaF and

Triclosan. For SM6 strain, 1 to 30% were similar

to the positive control. The 0% S-PRG toothpaste

presented no signicant

S. mutans

adherence

inhibition potential compared to negative control

(p>0.05).

Antibacterial potential of the toothpastes on

early-stage

S. mutans

biolm

The mean and standard deviation data for

the treatments applied after 4 h of reference and

the clinical

S. mutans

strains biolm formation

are shown in Figure 2a-2b, respectively. ANOVA

one-way indicated signicant difference for the

tested groups (p<0.05). According to Tukey’s

test results, all S-PRG containing toothpastes

were signicantly different from the negative

control for both strains (p<0.0001). The most

effective products were the 20% and 30% S-PRG

toothpastes. Nevertheless, all were signicantly

different from the positive commercial product

(NaF + Triclosan) that showed the higher

antimicrobial effect against early-stage biolms

(p<0.0001).

Figure 1 – Mean CFU/mL (log10) and standard deviation data when

treatments were applied previously to

S. mutans

adherence (before

bioﬁlm formation) over the UA159 reference (a) and SM6 clinical

(b) strains. Diﬀerent upper case letters mean signiﬁcant statistical

diﬀerences.

Braz Dent Sci 2023 Jan/Mar;26 (1): e3672

Spinola MS et al.

Antimicrobial efficacy of S-PRG containing toothpastes on S. mutans biofilm dev elopment

Spinola MS et al. Antimicrobial eﬃcacy of S-PRG containing toothpastes on

S. mutans bioﬁlm development

Antibacterial potential of the toothpastes on

mature

S. mutans

biolm

The mean and standard deviation data of cell

counts for the treatments applied after 24 h of

reference and clinical

S. mutans

biolm formation

are shown in Figure 3a-3b, respectively. ANOVA

one-way showed signicant difference for the

groups (p<0.05). According to the Tukey’s test,

there were no significant differences for the

S-PRG containing toothpastes compared to the

negative control, when reference UA159 strain

was tested (p>0.05). For the SM6 clinical strain,

the experimental toothpastes exhibited signicant

antimicrobial effect compared to the negative

control, with higher efcacy when 20 and 30%

S-PRG were used, but they were not superior than

the commercial NaF toothpaste used as positive

control (p<0.05).

DISCUSSION

The effect of toothpastes containing different

concentrations of S-PRG was evaluated at three

distinct moments: before, after 4 h, and after 24 h

of biolm development, to represent the different

clinical situations found in the oral cavity.

The ve minutes treatment time was chosen to

favour the effect of the toothpastes in this in vitro

model, since only one application was performed.

A standard strain of

S. mutans

(UA 159) was

used because this microorganism represents one

of the most important microorganisms in the

aetiology of caries disease [25]. Additionally, a

clinical strain, previously isolated from dental

caries (SM6), was used in order to extend the

knowledge of the antimicrobial properties of the

S-PRG toothpastes to a more real condition [26].

The model used here for biolm growth has been

widely used in the microbiology and cariology

areas [23,27] to simulate the conditions of

a cariogenic biofilm due to the presence of

5% sucrose added to the culture medium.

The biofilm was cultivated over bovine tooth

specimens. The use of bovine specimens in

substitution of human in caries lesion studies is

a viable alternative due to their similar mineral

content [28] and biolm cariogenicity pattern

with both substrates [29]. Besides, they present

great advantages, such as easier acquisition and

the presence of a large and at surface to obtain

the specimens.

Figure 2 – Mean CFU/mL (log10) and standard deviation data for

the antibacterial potential of treatments over the early-stage

S. mutans

bioﬁlms (after 4 h of bioﬁlm growth) cultivated from

UA159 reference (a) and SM6 clinical (b) strains. Diﬀerent upper

case letters mean signiﬁcant statistical diﬀerences.

Figure 3 – Mean CFU/mL (log10) and standard deviation data for

the antibacterial potential of treatments over

S. mutans

bioﬁlms

maturation (after 24 h of bioﬁlm growth) cultivated from UA159

reference (a) and SM6 clinical (b) strains. Diﬀerent upper case

letters mean signiﬁcant statistical diﬀerences.

Braz Dent Sci 2023 Jan/Mar;26 (1): e3672

Spinola MS et al.

Antimicrobial efficacy of S-PRG containing toothpastes on S. mutans biofilm dev elopment

Spinola MS et al. Antimicrobial eﬃcacy of S-PRG containing toothpastes on

S. mutans bioﬁlm development

The null hypothesis was rejected, as the

toothpastes containing S-PRG were able to inhibit

the adherence of the microorganisms and all the

concentrations tested reduced the development

of the early-stage biofilm. The toothpastes

containing 20% and 30% were superior to the

positive control in inhibiting biolm development

for the standard strain, while toothpastes from

1 – 30% were had similar effect to the positive

control in inhibiting biofilm development for

the clinical strain. These results agree with

previous studies, in which the ions released from

S-PRG in composite materials [13,30] and its

eluate [31,32] inhibited the growth of

S. mutans

biolm. Inhibition of initial microorganisms is

an important factor in prevention of the biolm

development because coaggregation inuences

the properties of plaque [33]. The observed

inhibition of biolm growth and effect on the

newly formed biolm are probably associated

with the release of BO3

3-, F- and Sr2+ as it has been

suggested that these ions are able to diffuse into

the biolm [34].

It is known that BO3

3- presents great

ability to prevent bacterial growth, because

it acts by inhibiting quorum sensing inside

bacteria [35]. Quorum sensing is characterized

as a highly specic system that controls different

bacterial activities such as the production of

signalling molecules, transport, and perception

of the surrounding environment. This system is

responsible for the process of bacterial adhesion

to surfaces, the production of the extracellular

polysaccharide matrix, and the virulence of the

biolm [36,37].

The F- ions are inhibitors of carbohydrate

metabolism in oral Streptococci [38] because

they can penetrate the cell and bind to enolase

and ATPase, inhibiting these enzymes and

leading to the reduction of carbohydrate

metabolism [39-41]. The inhibition of enolase

causes a reduction in phosphoenolpyruvate levels

and, consequently, a reduction in glucose uptake

by phosphoenolpyruvate phosphotransferase

system [42,43] while the inhibition of ATPase

causes ineffective extrusions of protons [44],

leading to an acidication of cell cytoplasm and

reduced metabolism and acid tolerance by the

cells [41,45].

Regarding the Sr2+ released from the S-PRG

fillers, the cariostatic activity caused by the

presence of such ions in oral products is not

completely understood, since it seems that its

concentration in saliva and plaque is usually

not able to exert an important antimicrobial

effect [46]. Nevertheless, the release of Sr2+

might represent an important ally in products

containing S-PRG, since its cariostatic properties

may act synergistically to F- to inhibit bacterial

activity [47].

In mature biolms (24h), S-PRG toothpastes

had no effect on the standard strain, but all

concentrations of S-PRG were able to decrease

the number of bacteria in the biolm cultivated

from the clinical strain SM6. Nevertheless,

this effect was lower than the NaF + triclosan

toothpaste. Triclosan, in the concentration of

0.3% in the dentifrice used as the positive control,

has well-established antimicrobial properties

when added to toothpastes to act against dental

plaque [48], although it has been replaced in

the recently launched toothpastes due to its

potential hormonal disfunction properties [49].

This active agent may be released in a higher

concentration than the ions present in the S-PRG

particle, to exert its antimicrobial effect on the

mature biolm, but this was not tested in the

present study.

It was previously shown that the inhibitory

effect of a S-PRG eluate was less pronounced when

the eluate was applied in the post-logarithmic

phase of biofilm growth, suggesting that the

main action of these bioactive particles is related

to the inhibition of

S. mutans

virulence and

growth [32]. The fact that S-PRG could decrease

the number of bacteria in the clinical strain, but

not in the standard strain may be related to the

fact that

S. mutans

standard strain UA 159 is

known to be a highly cariogenic strain, besides

being adapted to the culture medium since it is

the strain of choice for many studies in cariology.

The difference between the cariogenicity and

adaptation potential of the standard strain

UA159 and the clinical strain SM6 is clear when

we observed that for the three types of biolms

tested, the products presented better results with

the clinical strain.

These less favourable results observed

with the mature biolms can be explained by

the fact that the dental products containing

S-PRG particles, in general, are characterized as

agents that constantly release the ions present

in their composition. However, the amount of

ions released from the experimental toothpastes

Braz Dent Sci 2023 Jan/Mar;26 (1): e3672

Spinola MS et al.

Antimicrobial efficacy of S-PRG containing toothpastes on S. mutans biofilm dev elopment

Spinola MS et al. Antimicrobial eﬃcacy of S-PRG containing toothpastes on

S. mutans bioﬁlm development

might not be high enough to act in the rupture

of a mature biolm. In addition, the short time

of exposure to the treatments and the dilution

of the products in articial saliva prior to the

treatment of enamel specimens, to simulate

the dilution caused by saliva in the oral cavity,

may also have influenced the efficacy of the

products. Previous studies have shown that low

concentrations of S-PRG in eluates, for example,

were not effective on mature biolm and that

higher concentrations of these products were

needed to cause a rupture in the mature biolm

when compared to the concentration to inhibit

the biolm development [31]. This may be due

to the fact that the properties of a mature biolm

are completely different when compared to a

newly formed biolm. In a mature cariogenic

biofilm, formed by the interaction between

bacteria and sucrose, we can observe the presence

of an extracellular polysaccharide matrix that is

responsible for its virulence and that interferes

with the physical and chemical properties of the

biolm [50,51].

The presence of the extracellular matrix

promotes a support for the development of the

biolm because it encourages bacterial adhesion,

besides hindering the diffusion of substrates from

the medium to the interior of the biolm [50],

thus impairing the action of the ions inside the

biolm.

The results obtained with the experimental

toothpastes containing S-PRG are promising,

especially those with higher concentrations of

particles in their composition. Nevertheless, more

studies are needed to conrm the action of these

agents on the dental biolm in more relevant

clinical conditions.

CONCLUSION

Experimental dentifrices containing

S-PRG showed antimicrobial activity mainly on

microorganisms’ adherence and on early-stage

mutans

biolm, being promising agents to prevent

S. mutans

biolm growth and development.

Acknowledgements

The authors wish to express thanks to

Shofu Inc., Japan for providing the materials

for this study, and to CAPES and CNPq (processes

38882.434264/2019-01 and 51256/2019) for the

scholarships granted.

Author’s Contributions

MSP, JCJ, CRGT, ABB: Conceptualization.

MSP, JLM, MTG, JCJ: Methodology. MSP, MTG,

ABB: Formal Analysis. ABB: Resources. MSP, ABB:

Writing – Original Draft Preparation. MSP, TMFC,

JCJ, CRGT, ABB: Writing – Review & Editing.

ABB: Supervision. ABB: Project Administration.

Conict of Interest

No conicts of interest declared concerning

the publication of this article.

Funding

Part of this study was funded by CNPq

(process 51256/2019).

Regulatory Statement

The study protocol was approved by the local

Ethics Committee in Research and the approval

code for this study is: CAAE:005611/2017.

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Taciana Marco Ferraz Caneppele

(Corresponding address)

Departamento de Odontologia Restauradora, Instituto de Ciência e Tecnologia da

UNESP, São José dos Campos, Brasil.

Email: taciana.caneppele@unesp.br

Date submitted: 2022 Oct 02

Accept submission: 2023 Jan 21