Abstract This study evaluated the bioactive potential of a macro-porous chitosan scaffold incorporated with calcium hydroxide (CH-Ca) and functionalized with bioactive doses of simvastatin (SV) for bone tissue regeneration. Initially, the bioactive dose of SV in osteoblastic cells (SAOS-2) was determined. For the direct contact experiment, SAOS-2 cells were plated on scaffolds to assess cell viability and osteogenic differentiation. The second assay was performed at a distance using extracts from scaffolds incubated in culture medium to assess the effect of conditioned medium on viability and osteogenic differentiation. The initial screening showed that 1 μM SV presented the best biostimulating effects, and this dose was selected for incorporation into the CH-Ca and pure chitosan (CH) scaffolds. The cells remained viable throughout the direct contact experiment, with the greatest cell density in the CH-Ca and CH-Ca-SV scaffolds because of their higher porosity. The CH-Ca-SV scaffold showed the most intense bio-stimulating effect in assays in the presence and absence of osteogenic medium, leading to an increased deposition of mineralized matrix. There was an increase in the viability of cells exposed to the extracts for CH-Ca, CH-SV, and CH-Ca-SV during the one-day period. There was an increase in ALP activity in the CH-Ca and CH-Ca-SV; however, the CH-Ca-SV scaffold resulted in an intense increase in the deposition of mineralized nodules, approximately 56.4% at 7 days and 117% at 14 days, compared with CH (control). In conclusion, functionalization of the CH-Ca scaffold with SV promoted an increase in bioactivity, presenting a promising option for bone tissue regeneration.

Resumo Este trabalho teve como objetivo avaliar a influência da interface de uma restauração adesiva na difusão do peróxido de hidrogênio (H2O2), toxicidade indireta e expressão de mediadores pró-inflamatórios por células odontoblastóides, após clareamento dental em consultório. Cavidades dentárias preparadas em discos de esmalte / dentina foram restauradas com adesivo e submetidas ou não à degradação hidrolítica (HD). Um gel clareador com 35% H2O2 (WG) foi aplicado por 45 min em discos restaurados e não restaurados adaptados às câmaras pulpares artificiais dando origem aos grupos: SD- discos intactos (controle); SD / HP - Discos intactos clareados; RT / HP - discos restaurados e clareados; e RT / HD / HP - discos restaurados, clareados e submetidos a HD. Os extratos (meio de cultura + componentes WG difundidos através da interface esmalte/dentina/restauração) foram coletados e aplicados em células odontoblastóides MDPC-23. Foi avaliada a quantidade de H2O2 nos extratos, bem como a viabilidade (CV), morfologia (CM) e expressão gênica de mediadores inflamatórios (TNF-α e COX-2) pelas células pulpares expostas aos extratos (ANOVA e testes de Tukey; 5% de significância). Todos os grupos clareados apresentaram menor CV do que SD (controle; p <0,05). A maior redução CV e expressão gênica de TNF-α e COX-2 foi observada no grupo RT / HD / HP em comparação com SD / HP e RT / HP (controle; p <0,05). Alterações na CM ocorreram em todos os grupos clareados. A intensidade desses efeitos celulares teve relação direta com a quantidade de H2O2 nos extratos. Concluímos que a presença de uma cavidade contendo restauração adesiva aumenta a difusão de H2O2 após o clareamento em consultório, o que, por sua vez, aumenta a toxicidade indireta dessa terapia e desencadeia a expressão de mediadores pró-inflamatórios pelas células pulpares MDPC-23.

Abstract This paper aimed to assess the influence of adhesive restoration interface on the diffusion of hydrogen peroxide (H2O2), indirect toxicity, and pro-inflammatory mediators expression by odontoblast-like cells, after in-office tooth whitening. Dental cavities prepared in bovine enamel/dentin discs were adhesively restored and subjected or not to hydrolytic degradation (HD). A whitening gel with 35% H2O2 (WG) was applied for 45 min onto restored and non-restored specimens adapted to artificial pulp chambers giving rise to the groups: SD- intact discs (control); SD/HP- whitened intact discs; RT/HP- restored and whitened discs; and RT/HD/HP- restored and whitened discs subjected to HD. The extracts (culture medium + WG components diffused through enamel/dentin/restoration interface) were collected and applied to odontoblast-like MDPC-23 cells. The study evaluated the amount of H2O2 in the extracts, as well as the cell viability (CV), cell morphology (CM), and gene expression of inflammatory mediators (TNF-α and COX-2) by the pulp cells exposed to the extracts (ANOVA and Tukey tests; 5% significance). All whitened groups presented lower CV than SD (control; p<0.05). The highest CV reduction and gene expression of TNF-α and COX-2 was observed in the RT/HD/HP group in comparison with SD/HP and RT/HP (control; p<0.05). CM alterations occurred in all whitened groups. The intensity of these cell side effects was directly related with the amount of H2O2 in the extracts. We concluded that adhesive restoration of dental cavity increases the H2O2 diffusion after in-office whitening, enhancing the indirect toxicity of this therapy and trigger pro-inflammatory overexpression by MDPC-23 cells.

Abstract Objective The objective of this study was to evaluate dental sensitivity using visual analogue scale, a Computerized Visual Analogue Scale (CoVAS) and a neurosensory analyzer (TSA II) during at-home bleaching with 10% carbamide peroxide, with and without potassium oxalate. Materials and Methods Power Bleaching 10% containing potassium oxalate was used on one maxillary hemi-arch of the 25 volunteers, and Opalescence 10% was used on the opposite hemi-arch. Bleaching agents were used daily for 3 weeks. Analysis was performed before treatment, 24 hours later, 7, 14, and 21 days after the start of the treatment, and 7 days after its conclusion. The spontaneous tooth sensitivity was evaluated using the visual analogue scale and the sensitivity caused by a continuous 0°C stimulus was analyzed using CoVAS. The cold sensation threshold was also analyzed using the TSA II. The temperatures obtained were statistically analyzed using ANOVA and Tukey's test (α=5%). Results The data obtained with the other methods were also analyzed. 24 hours, 7 and 14 days before the beginning of the treatment, over 20% of the teeth presented spontaneous sensitivity, the normal condition was restored after the end of the treatment. Regarding the cold sensation temperatures, both products sensitized the teeth (p<0.05) and no differences were detected between the products in each period (p>0.05). In addition, when they were compared using CoVAS, Power Bleaching caused the highest levels of sensitivity in all study periods, with the exception of the 14th day of treatment. Conclusion We concluded that the bleaching treatment sensitized the teeth and the product with potassium oxalate was not able to modulate tooth sensitivity.

Abstract Objective This study verified the occurrence of dental sensitivity in patients submitted to a 35% hydrogen peroxide based product (Whiteness HP Maxx 35% – FGM), skin cold sensation threshold (SCST) and its influence on dental sensitivity. Material and Methods Sixty volunteers were divided into 4 groups (n = 15), according to SCST (low: GI and GIII, and high: GII and IV) and bleaching treatment (hydrogen peroxide: GI and GII, and placebo: GIII and GIV). SCST was determined in the inner forearm for 6 different times using a neurosensory analyzer, the TSA II (Medoc Advanced Medical Systems, Ramat Yishai, Northern District, Israel). Dental sensitivity measurements were performed 10 different times using a thermal stimulus and an intraoral device attached to TSA II, positioned in the buccal surface of the upper right central incisor. Spontaneous dental sensitivity was also determined using the Visual Analogue Scale (VAS). Data were submitted to Student's t-test and Pearson's Correlation Test (α=0.05). SCST remained the same during bleaching treatment. Results Distinct responses of dental sensitivity were found in patients with low and high SCST during the first and third bleaching session (p≤0.05). The teeth submitted to the bleaching treatment became more sensitive to cold than those treated with placebo. Moreover, data obtained with TSA and VAS presented moderate correlation. Conclusions Bleaching treatment increased dental sensitivity and skin cold sensation threshold might represent a determining factor in this occurrence, since low and high SCST patients had different responses to the thermal stimulus in the teeth.

Resumo Este estudo teve como objetivo avaliar a penetração trans-amelodentinária do peróxido de hidrogênio (H2O2) aplicados em dentes bovinos pigmentados com chá preto em diferentes intensidades. Divisão dos grupos: AD em água destilada; CP100 em uma infusão de 1,6 g de chá preto para 100 mL de água destilada; CP10 em uma infusão de 1,6 g de chá preto para 10 mL de água destilada. Todos os grupos foram imersos por 6 dias. Para quantificar a penetração de H2O2, as amostras foram colocadas em câmaras pulpares artificiais (CPAs) e submetidas a um tratamento clareador com PH a 38%, uma vez por semana durante 3 semanas. Após o tratamento clareador, a solução tampão de acetato das CPAs com a enzima da peroxidase, foi avaliada num espectrofotômetro de reflexão. A penetração trans-amelodentinária de PH e os valores de L* obtidos em T1, T2 e T3 foram submetidos ao teste estatístico de Kruskal-Wallis e Friedman. Em T1, a difusão de H2O2 no AD foi mais elevada do que em CP100 e CP10. Nos outros tempos de avaliação, os valores de penetração no CP100 e CP10 aumentaram e permaneceram semelhantes. Os valores L* aumentaram significativamente em todos os grupos no T1. No T2, os valores L* foram maiores no AD e os valores em CP100 e CP10 foram semelhantes entre si. No último tempo, o CP10 apresentou os menores valores de L*. Diferentes níveis de pigmentação não afetaram a penetração de H2O2 através do esmalte e dentina e o agente de clareador foi eficaz na alteração cromática.

Abastract This study aimed to evaluate the transenamel and transdentinal penetration of hydrogen peroxide (H202) applied to bovine teeth pigmented with black tea at different intensities. The following groups were formed DW: immersion in distilled water; BT100: immersion in an infusion of 1.6 g of black tea per 100 mL distilled water; BT10: immersion in an infusion of 1.6 g black tea per 10 mL distilled water. All groups were immersed for 6 days. To quantify the penetration of H202, the specimens were placed in artificial pulp chambers (APCs) and subjected to bleaching treatment with 38% H2O2 once per week for 3 weeks. After bleaching treatment, the acetate buffer solution of APCs with peroxidase enzyme was evaluated in a reflection spectrophotometer. The transenamel and transdentinal penetration of H2O2 and the L* values obtained at T1, T2 and T3 were subjected to Kruskal-Wallis and Friedman statistical tests. At T1, the H2O2 diffusion in DW was higher than that in BT100 and BT10. At the other evaluation times, the penetration values in BT100 and BT10 increased and remained similar. The L* values increased significantly in all groups at T1. At T2, the L* values were higher in DW, while the values in BT100 and BT10 were similar to each other. At the end of the experiment, BT10 showed the lowest L* values. The pigmentation level did not affect the penetration of H2O2 through the enamel and dentin and the bleaching agent effectively changed the color of the teeth.

Resumo O peróxido de hidrogênio (H2O2) é capaz de penetrar pelos tecidos dentários, alterando a coloração destes, e causar danos a polpa. Este estudo avaliou a penetração por esmalte e dentina, a alteração de cor e a reposta tecidual pulpar, provocadas pelo uso de duas concentrações de H2O2 em protocolos de clareação dentária de consultório. Discos de dentes bovinos em câmaras pulpares artificiais receberam géis clareadores para avaliação da penetração por esmalte e dentina e da alteração de cor, formando os grupos: BLU (H2O2 20% - 1x50 min, Whiteness HP Blue); MAX (H2O2 35% - 3x15 min, Whiteness HP Maxx); e Controle (gel placebo - 1x50 min). A penetração por esmalte e dentina foi quantificada baseada na reação do H2O2 com o corante violeta leucocristal, e a alteração de cor foi analisada pelo sistema CIELab. Vinte ratos Wistar foram divididos em dois grupos (BLU e MAX), e tiveram os molares direito superiores tratados com os mesmos protocolos do estudo in vitro; os molares superiores do lado esquerdo serviram de controle. Após 2 dias, os animais foram eutanasiados e as maxilas examinadas por microscopia de luz. Foram atribuídos escores ao infiltrado inflamatório (1, ausente; 2, leve; 3, moderado; 4 severo ou necrose). Os dados foram submetidos a testes estatísticos (=0,05). O grupo MAX apresentou maior penetração de H2O2 por esmalte e dentina (p<0,05). A alteração de cor foi semelhante nos grupos clareados (p>0,05), mas diferente quando comparados grupos clareados com controle (p<0,05). MAX apresentou inflamação severa nos terços superiores da polpa coronária, e BLU apresentou inflamação moderada (p<0,05). Assim, protocolo para procedimento clareador de consultório utilizando baixas concentrações de H2O2 deve ser de escolha na clínica, por reduzir a penetração por esmalte e dentina, causando menos danos à polpa, e proporcionar mesma eficiência clareadora.

Abstract Hydrogen peroxide (H2O2) penetrates into the dental hard tissues causing color alteration but also alterations in pulpal tissues. Hard-tissue penetration, color alteration and the pulp response alterations were evaluated for two in-office bleaching protocols with H2O2. For trans-enamel/dentin penetration and color alteration, discs of bovine teeth were attached to an artificial pulp chamber and bleached according to the groups: BLU (20% H2O2 - 1x50 min, Whiteness HP Blue); MAX (35% H2O2 - 3x15 min, Whiteness HP Maxx); Control (1x50 min, placebo). Trans-enamel/dentin penetration was quantified based on the reaction of H2O2 with leucocrystal violet and the color analyzed by CIELab System. Twenty Wistar rats were divided into two groups (BLU and MAX) and their maxillary right molars were treated according to the same protocols of the in vitro study; the maxillary left molars were used as controls. After 2 days, the animals were killed and their maxillae were examined by light microscopy. The inflammation of pulp tissue was scored according to the inflammatory infiltrate (1, absent; 2, mild; 3, moderate; 4, severe/necrosis). Data were analyzed by statistical tests (α=0.05). MAX showed higher trans-enamel/dentinal penetration of H2O2 (p<0.05). The color alteration was similar for both groups (p>0.05), and different when compared to Control group (p<0.05). MAX showed severe inflammation in the upper thirds of the coronal pulp, and BLU showed moderate inflammation (p<0.05). In-office bleaching protocols using lower concentrations of hydrogen peroxide should be preferred due to their reduced trans-enamel/dentinal penetration since they cause less pulp damage and provide same bleaching efficiency.

<p>Este estudo avaliou a alteração de cor, a citotoxicidade e a difusão de peroxido de hidrogênio ocorridos em diferentes protocolos clareadores caseiros: peróxido de carbamida (PC) 10% por 3 ou 1,5 h; peróxido de hidrogênio (PH) 6% por 1,5 h ou 45 min. Para a quantificação da penetração do peróxido, discos de dentes bovinos foram posicionados em câmaras pulpares artificiais (CPAs) contendo solução tampão de acetato, que foi coletada para avaliação em espectrofotômetro. Para análise da citotoxicidade, os espécimes foram adaptados nas CPAs contendo meio de cultura, que posteriormente foi aplicado sobre células odontoblastóides MDPC-23 por 1 h. O metabolismo celular foi avaliado pelo teste MTT e a alteração de cor dos espécimes foi analisada pelo sistema CIE L*a*b*. Os dados foram submetidos a ANOVA e teste de Fisher (α=5). O tratamento com PC10% por 3 horas foi o mais efetivo, enquanto que o tratamento com PH 6% por 45 min produziu a menor alteração cromática. Os grupos PC 10% por 1,5 h e PH 6% por 45 min causaram a menor penetração trans-amelodentinária do peróxido, e PH 6% por 1,5 h, a maior difusão. Nenhum tratamento alterou o metabolismo celular. A redução do tempo de exposição aos peróxidos comprometeu o resultado clareador; maior penetração de peroxido não significa maior efetividade clareadora.</p>

<p>This study evaluated the color change, cytotoxicity and hydrogen peroxide (HP) diffusion caused by different home bleaching protocols: 10% carbamide peroxide (CP) for 3 or 1.5 h, 6% hydrogen peroxide for 1.5 h or 45 min. To quantify the peroxide penetration, disks of bovine teeth were placed in artificial pulp chambers (APCs) containing acetate buffer, which was collected for evaluation in a spectrophotometer. For analysis of cytotoxicity, specimens were adapted in APCs containing culture medium, which subsequently was applied on MDPC-23 odontoblast-like cells for 1 h. Cellular metabolism was evaluated by methyl tetrazolium (MTT) assay and the color change of the specimens was analyzed using the CIE L * a * b * system. The data were submitted to ANOVA and Fisher test (α=5%). The treatment with 10% CP for 3 h was the most effective, and 6% HP for 45 min produced the lowest color change. The groups 10% CP for 1.5 h and 6% HP for 45 min had the lowest trans-enamel dentinal HP penetration, and the 6% HP for 1.5 h had the highest. None of the protocols affected cellular metabolism and morphology. In conclusion, reduced peroxide exposure time reduced the bleaching result; higher HP diffusion did not mean higher effectiveness.</p>

O objetivo deste estudo foi avaliar a desmineralização e a penetração do peróxido de hidrogênio (HP) em dentes com lesões incipientes submetidos ao tratamento clareador. Para analisar a penetração do peróxido de hidrogênio, discos de esmalte/dentina hígidos e desmineralizados foram posicionados em câmaras pulpares artificiais contendo solução tampão de acetato. Após o tratamento clareador, esta solução foi submetida à análise da densidade óptica no espectrofotômetro e as superfícies dos discos foram analisadas por meio de microscopia eletrônica de varredura (MEV) e microscopia de luz polarizada (MLP). Os discos restantes foram submetidos à análise de microdureza transversal em diferentes profundidades. Os dados foram submetidos aos testes ANOVA e teste PLSD Fisher (= 5%). Observou-se que os dentes previamente desmineralizados mostraram maior penetração de HP (p<0,05). O tratamento clareador causou alterações em uma profundidade de 20 μm em esmalte hígido e até 90 μm em esmalte desmineralizado. As imagens obtidas em PLM e MEV mostraram que o tratamento clareador promove alterações superficiais no esmalte, sendo mais pronunciadas em dentes previamente desmineralizados. Foi concluído que o nível de mineralização do esmalte influencia a penetração do PH e que o agente clareador contribuiu para o aumento da profundidade de desmineralização.

The aim of this study was to evaluate the demineralization and hydrogen peroxide (HP) penetration in teeth with incipient lesions submitted to bleaching treatment. For analysis of HP penetration, sound and demineralized enamel/dentin discs were placed in artificial pulp chambers containing acetate buffer solution. After bleaching treatment, this solution was subjected for analysis of optical density by spectrophotometry and the disc surfaces were analyzed with scanning electron microscopy (SEM) and polarized light microscopy (PLM). The remaining discs were subjected for cross-sectional hardness analysis at different depths. Data were analyzed by repeated measures ANOVA and PLSD Fisher test (a=0.05). It was observed that previously demineralized teeth showed greater HP penetration (p<0.05). The bleaching treatment caused changes to a depth of 20 µm in sound enamel and up to 90 µm in demineralized enamel. SEM and PLM images revealed that the bleaching treatment caused superficial changes that were considerably more accentuated in previously demineralized teeth. It may be concluded that the enamel mineralization level influences HP penetration and the bleaching agent contributed to increase the demineralization depth.