Establishment of a saliva donor selection for in vitro biofilm growth

Resumo Introdução O emprego de biofilmes polimicrobianos, utilizando a saliva como inóculo, é um modelo promissor para o estudo de biofilmes cariogênicos in vitro. Entretanto, ainda não existe uma padronização para seleção de doadores de saliva. Objetivo O objetivo deste estudo foi estabelecer uma metodologia para seleção de doadores de saliva utilizando fatores salivares microbianos e características in vitro do biofilme. Material e método Para doação de saliva foram selecionados vinte voluntários. Os voluntários permaneceram 24 horas sem escovar os dentes e ficaram em jejum por 2 horas antes da coleta da saliva. Foram avaliados os seguintes parâmetros: viabilidade das bactérias anaeróbias totais e mutans streptococci; concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM) da clorexidina; capacidade de formação de biofilme por meio da biomassa; e a suscetibilidade dos biofilmes à clorexidina. Resultado A viabilidade bacteriana da saliva, a capacidade de formação de biofilme e a suscetibilidade do biofilme à clorexidina foram apresentadas como média e intervalo de confiança (95%). A diferença entre a viabilidade do biofilme (mutans streptococci e bactérias totais) após tratamento com NaCl 0,9% e diacetato de clorexidina 0,2% foi comparada pelo teste t de Student com nível de significância estabelecido em 5%. A viabilidade total de bactérias anaeróbias (mediana) foi de 7,28 log 1+UFC/mL (unidades formadoras de colônia/mL). A viabilidade dos mutans streptococci na saliva apresentou mediana de 5,47 log 1+UFC/mL. Para capacidade de formação de biofilme a mediana da biomassa foi de 0,1172 A570. Conclusão O tratamento com clorexidina reduziu significativamente os mutans streptococci e a viabilidade total das bactérias. A metodologia para seleção do doador de saliva foi estabelecida com sucesso

In vitro effect of low-fluoride toothpaste supplemented with sodium trimetaphosphate, xylitol, and erythritol on enamel demineralization

Regular use of toothpaste with fluoride (F) concentrations of ≥ 1000 ppm has been shown to contribute to reducing caries increment. However, when used by children during the period of dental development, it can lead to dental fluorosis. Objective: In this study, we aimed to evaluate the in vitro effect of a toothpaste formulation with reduced fluoride (F) concentration (200 ppm) supplemented with sodium trimetaphosphate (TMP: 0.2%), Xylitol (X:16%), and Erythritol (E: 4%) on dental enamel demineralization. Methodology: Bovine enamel blocks were selected according to initial surface hardness (SHi) and then divided into seven experimental toothpaste groups (n=12). These groups included 1) no F-TMP-X-E (Placebo); 2) 16% Xylitol and 4% Erythritol (X-E); 3) 16% Xylitol, 4% Erythritol and 0.2%TMP (X-E-TMP); 4) 200 ppm F (no X-E-TMP: (200F)); 5) 200 ppm F and 0.2% TMP (200F-TMP); 200 ppm F, 16% Xylitol, 4% Erythritol, and 0.2% TMP (200F-X-E-TMP); and 7) 1,100 ppm F (1100F). Blocks were individually treated 2×/day with slurries of toothpastes and subjected to a pH cycling regimen for five days (DES: 6 hours and RE: 18 hours). Then, the percentage of surface hardness loss (%SH), integrated loss of subsurface hardness (ΔKHN), fluoride (F), calcium (Ca), and phosphorus (P) in enamel were determined. The data were analyzed by ANOVA (1-criterion) and the Student-Newman-Keuls test (p<0.001). Results: We found that the 200F-X-E-TMP treatment reduced %SH by 43% compared to the 1100F treatments (p<0.001). The ΔKHN was ~ 65% higher with 200F-X-E-TMP compared to 1100F (p<0.001). The highest concentration of F in enamel was observed on the 1100F treatment (p<0.001). The 200F-X-E-TMP treatment promote higher increase of Ca and P concentration in the enamel (p<0.001). Conclusion: The association of 200F-X-E-TMP led to a significant increase of the protective effect on enamel demineralization compared to the 1100F toothpast

Effect of fluoride gels with nano-sized sodium trimetaphosphate on the in vitro remineralization of caries lesions

Objective: To evaluate the effects of fluoride (F) gels supplemented with micrometric or nano-sized sodium trimetaphosphate (TMPmicro and TMPnano, respectively) on the in vitro remineralization of caries-like lesions. Methodology: Bovine enamel subsurface lesions (n=168) were selected according to their surface hardness (SH) and randomly divided into seven groups (n=24/group): Placebo (without F/TMP), 4,500 ppm F (4500F), 4500F + 2.5% TMPnano (2.5% Nano), 4500F + 5% TMPnano (5% Nano), 4500F + 5% TMPmicro (5% Micro), 9,000 ppm F (9000F), and 12,300 ppm F (Acid gel). The gels were applied in a thin layer for one minute. Half of the blocks were subjected to pH cycling for six days, whereas the remaining specimens were used for loosely- (calcium fluoride; CaF2) and firmly-bound (fluorapatite; FA) fluoride analysis. The percentage of surface hardness recovery (%SHR), area of subsurface lesion (ΔKHN), CaF2, FA, calcium (Ca), and phosphorus (P) on/in enamel were determined. Data (log10-transformed) were subjected to ANOVA and the Student-Newman-Keuls’ test (p<0.05). Results: We observed a dose-response relation between F concentrations in the gels without TMP for %SHR and ΔKHN. The 2.5% Nano and 5% Micro reached similar %SHR when compared with 9000F and Acid gels. For ΔKHN, Placebo and 5% Nano gels had the highest values, and 5% Micro, 2.5% Nano, 9000F, and Acid gels, the lowest. All groups had similar retained CaF2 values, except for Placebo and Acid gel. We verified observed an increase in Ca concentrations in nano-sized TMP groups. Regarding P, TMP groups showed similar formation and retention to 9000F and Acid. Conclusion: Adding 2.5% nano-sized or 5% micrometric TMP to low-fluoride gels lead to enhanced in vitro remineralization of artificial caries lesions

Effect of treatment with phosphate, casein phosphopeptide and fluoride on the remineralization: in vitro study

Abstract This study aimed to evaluate in vitro the effect protocols and anticaries agents containing casein amorphous calcium fluoride phosphopeptide-phosphate (CPP-ACPF, MI Paste Plus), sodium trimetaphosphate (TMP) and fluoride (F), in remineralization of caries lesions. Bovine enamel blocks with initial caries lesions were divided into groups (n = 12): 1) Toothpaste without F-TMP-MI Plus (Placebo); 2) Toothpaste 1100 ppm F (1100F), 3) 1100F + MI Paste Plus (1100F-MI Paste Plus), 4) Toothpaste with 1100F + Neutral gel with 4,500 ppm F + 5%TMP (1100F + Gel TMP) and 5) Toothpaste with 1100F + Neutral gel with 9,000 ppm F (1100F + Gel F). For the 4 and 5 groups the gel was applied only once for 1 minute, initially to the study. For the 3 group, after treatment with 1100F, MI Paste Plus was applied 2x/day for 3 minute. After pH cycling, the percentage of surface hardness recovery (%SHR); integrated loss of subsurface hardness (ΔKHN); profile and depth of the subsuperficial lesion (PLM); concentrations of F, calcium (Ca) and phosphorus (P) in enamel was determined. The data were analyzed by ANOVA (1-criterion) and Student-Newman-Keuls test (p 0.001). Similar values of P in the enamel were observed in the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001), except for the 1100F + Gel TMP group, which presented a high concentration (p < 0.001). We conclude that the 1100F+TMP gel treatment/protocol led to a significant increased remineralization when compared to the other treatments/protocols and may be a promising strategy for patients with early caries lesions

Efeito de dentifrícios fluoretados e suplementados com nanopartículas de trimetafosfato de sódio sobre a desmineralização, remineralização e erosão dentária

The aim of this study was to evaluate the ability of conventional toothpaste (1100 ppm F) supplemented with micrometric or nano-sized sodium trimetaphosphate (TMP; TMPnano ), in reducing demineralization (in vitro), promote remineralization ( in situ ) and reduce erosion tooth (in vitro). In the study of demineralization of bovine enamel blocks (n = 96 ) selected by the initial surface hardness ( SHi ) were divided into eight groups of toothpaste (n = 12) without fluoride and without TMP (Placebo), 1100 ppm F (1100 ppm F) 1100 ppm F associated 1% TMP micrometric and nano-sized (1100 1%TMP; 1100 1%TMPnano), 1100 ppm F associated with 3% TMP micrometric and nano-sized (1100 3%TMP; 1100 3%TMPnano), 1110 ppm F associated with 6% TMP micrometric and nano-sized TMP (1100 6%TMP; 1100 6%TMPnano). The blocks were subjected to pH cycling for five days, and treatment with their toothpastes made 2x/day. Next, we determined the final surface hardness (SHf), integrated mineral loss (IML), differential integrated mineral loss (ΔIML) and fluoride (F) in the enamel. The results were subjected to analysis of variance followed by Student-Newman-Keuls test (p < 0.001). The group 1100 3 %TMPnano had the lowest mineral loss (SHF, IML and ΔIML) followed by 1100 3%TMP group (p < 0.001). The group 100 3%TMPnano showed a higher concentration of F in the enamel, followed by 1100 6%TMPnano (p < 0.001). For the study of remineralization of bovine enamel blocks (n = 192) were selected by the hardness of the surface after demineralization (SH1), and divided into four groups: Placebo, 1100 ppm F, 1100 3 % TMP micrometric (1100 TMP) and 1100 3% TMP nano-sized (1100 TMPnano ). Twelve volunteers wore palatal appliances with four blocks of demineralized enamel, for three days, and brushing held 3x/day. After the period of remineralization...O objetivo deste estudo foi avaliar a capacidade de dentifrícios convencionais (1100 ppm F) suplementados com trimetafosfato de sódio micrométrico ou nanoparticulado (TMP; TMPnano), em reduzir a desmineralização (in vitro), promover a remineralização (in situ) e reduzir a erosão dentária (in vitro). No estudo de desmineralização, blocos de esmalte bovino (n = 96) selecionados pela dureza de superfície inicial (SHi) foram divididos em oito grupos de dentifrícios (n = 12): sem fluoreto e sem TMP (Placebo); 1100 ppm F (1100 ppm F); 1100 ppm F associado 1% TMP micrométrico e nanoparticulado (1100 1%TMP; 1100 1%TMPnano), 1100 ppm F associado a 3% TMP micrométrico e nanoparticulado (1100 3%TMP; 1100 3%TMPnano), 1110 ppm F associado a 6% TMP micrométrico e nanoparticulado (1100 6%TMP; 1100 6%TMPnano). Os blocos foram submetidos a ciclagem de pH durante cinco dias, sendo o tratamento com os respectivos dentifrícios realizados 2x/dia. A seguir, determinou-se a dureza de superfície final (SHf), perda mineral integrada (IML), diferencial da perda mineral integrada (ΔIML) e fluoreto (F) no esmalte. Os resultados foram submetidos à análise de variância seguida pelo teste Student-Newman-Keuls (p < 0,001). O grupo 1100 3%TMPnano apresentou a menor perda mineral (SHf, IML e ΔIML) seguido pelo grupo 1100 3%TMP (p < 0,001). O grupo 1100 3%TMPnano apresentou a maior concentração de F no esmalte, seguido pelo 1100 6%TMPnano (p < 0,001). Para o estudo de remineralização, blocos de esmalte bovinos (n = 192) foram selecionados pela dureza de superfície pós-desmineralização (SH1), e divididos em quatro grupos experimentais: Placebo, 1100 ppm F, 1100 3% TMP micrométrico (1100 TMP), 1100 3% TMP nanoparticulado (1100 TMPnano). Doze voluntários utilizaram dispositivos palatinos, com quatro blocos de esmalte..

Desenvolvimento de gel tópico com reduzida concentração de fluoreto: capacidade de inibir a desmineralização e promover a remineralização do esmalte dentário

O objetivo foi avaliar a capacidade de géis com baixa concentração de F suplementados com trimetafosfato de sódio (TMP), em reduzir a desmineralização (in vitro) e promover a remineralização (in situ) de esmalte dentário. No estudo in vitro, blocos de esmalte bovino (n = 160) selecionados pela dureza de superfície (SHi) foram divididos em oito grupos de géis (n=20): sem F e TMP (Placebo); TMP3%, TMP5%; 4500 μg F/g (4500); 4500 μg F/g + TMP3% (4500 TMP3%), 4500 μg F/g + TMP5% (4500 TMP5%), 9000 μg F/g (9000) e 12300 μg F/g (Gel ácido). Os blocos foram submetidos a ciclagem de pH durante cinco dias, após aplicação tópica dos géis. A seguir, determinou-se a dureza de superfície final (SHf), perda integrada de dureza de subsuperfície (ΔKHN), CaF2, F, Ca, P formados e retidos no esmalte. Os resultados foram submetidos à análise de variância e teste de Bonferroni (p0,05). O Gel ácido apresentou a maior concentração de CaF2 formado. O grupo 4500 TMP5% apresentou o maior valor de F formado (p0,05). O P formado foi semelhante entre os grupos (p>0,05) e o Gel ácido apresentou a maior concentração de P retido no esmalte. Concluiu-se que é possível inibir a desmineralização do esmalte com gel fluoretado de baixa concentração suplementado-o com TMP. Para o estudo in situ, blocos de esmalte bovinos (n=240) foram selecionados pela dureza de superfície (SH1), após desmineralização, e divididos em cinco grupos experimentais: Placebo, 4500, 4500 TMP5%, 9000 e Gel ácido. Doze voluntários utilizaram dispositivos palatinos, com quatro blocos de esmalte desmineralizados, durante três...The aim of this study was to evaluate the capacity of gels with low concentration of F supplemented with sodium trimetaphosphate (TMP) to reduce the dental enamel demineralization (in vitro) and remineralization (in situ). In the in vitro study blocks of bovine enamel (n=160) were selected through surface hardness (SHi) and divided into eight groups (n = 20): gel without F and TMP (Placebo), TMP3%, TMP5%, 4500 μg F/g (4500), 4500 μg F/g + TMP3% (4500 TMP3%), 4500 μg F/g + TMP5% (4500 TMP5%), 9000 μg F/g (9000) and 12.300 μg F/g (acid gel). The blocks were submitted to pH cycling during five days after topical application of gels. After that, the surface hardness (SHf), the subsurface hardness integrated loss (ΔKHN), CaF2, F, Ca and P formed and retained in enamel were determined. The results were submitted to analysis of variance and Bonferroni tests (p 0.05). The P formed was similar between the groups (p> 0.05) and the acid gel showed the highest concentration of P retained in enamel. It was concluded that it is possible to inhibit enamel demineralization with low F concentration gel by supplementing it with TMP 5%. In the in situ study bovine enamel blocks (n=240) were selected through surface hardness (SH1) after demineralization and divided into five groups: Placebo, 4500, 4500 TMP5%, 9000 and Acid gel). Twelve volunteers wore palatal appliances with four demineralized blocks, during three days after topical application of F (ATF). Two blocks were removed immediately after the ATF to ... (Complete abstract click electronic access below)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Application of a projective method for the assessment of children’s dental fear

The aims of this study were to assess children's dental fear by using a projective technique CDFP (Children’s dental fear picture test). 40 patients from a pediatric dental clinic, between 5-9 years old, took part in this study. At first, the CDFP was applied to the children. This method has three subtests and its overall assessment classifies children as "fearful, non fearful and uncertain". The children answered the CFSS-DS (Children Fear Survey Schedule–Dental Subscale). The results showed that CDFP has inter-examiner reliability and a significant correlation with the CFSS-DS was observed