Avaliação in vivo e in vitro do Biosilicato® frente ao estresse oxidativo, penetração do peróxido de hidrogênio e eficácia clareadora.
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Data
2019-06-14
Autores
Carminatti, Marina [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Novos produtos estão sendo desenvolvidos com o propósito de atenuar os efeitos dos compostos utilizados em procedimentos clareadores. O objetivo do presente estudo foi avaliar in vivo o potencial terapêutico do gel de Biosilicato® (BS) sobre o tecido pulpar de molares de ratos Wistar, por meio das análises histológica e imunoistoquímica, assim como analisar in vitro, a penetração do H2O2 e a eficácia clareadora em dentes bovinos. Para isso, este estudo foi dividido em 2 partes. Parte 1: Segmento in vivo: 50 hemi-maxilas de 25 ratos foram divididas aleatoriamente em 5 grupos (n=10): Controle- não recebeu qualquer tratamento; CLA- recebeu 30 min do gel clareador H2O2 35%; BS-CLA- recebeu 20 min do gel de BS, seguido de 30 min do H2O2 35%; CLA+BS- recebeu 30 min de uma mistura do H2O2 35% com o gel BS (1:1); CLA+H2O- recebeu 30 min de uma mistura do H2O2 35% com água destilada (1:1). Após 2 dias, os animais foram mortos e as hemi-maxilas separadas para análise histológica em H.E. Foram atribuídos escores à inflamação e os dados submetidos aos testes de Kruskal-Wallis e Dunn e teste de Mann-Whitney (p<0,05). Segmento in vitro: 50 discos de dentes bovinos foram acoplados em câmaras pulpares artificiais, e divididos em 5 grupos (n=10) que receberam os mesmos tratamentos que o segmento in vivo. A penetração do H2O2 por esmalte e dentina foi quantificada baseada na reação deste com o corante violeta leucocristal. A alteração de cor foi analisada pelo sistema CIELab, nos períodos: antes da sessão clareadora – (T0), imediatamente após (T1), e 24 horas após (T2), e os dados submetidos à análise de variância (ANOVA) seguida do teste Tukey (p<0,05). Parte 2: 40 hemi-maxilas de 20 ratos foram divididas aleatoriamente em 4 grupos (n=10): Controle, CLA, BS-CLA e CLA+BS. Após 2 dias, os animais foram mortos e as hemi-maxilas separadas para análise histológica em H.E. e imunoistoquímica para PCNA, HO-1, TNF-α e Substância P. Foram atribuídos escores à inflamação e à imunomarcação de TNF-α, e submetidos aos testes de Kruskal-Wallis e Dunn (p<0,05). Para análise da área correspondente à imunomarcação obtida através da densidade óptica de imunomarcação da Substância P foi realizada a análise da variância (ANOVA) seguida do teste de Tukey para comparação entre as médias. Foram realizadas as contagens das células imunomarcadas para HO-1 e PCNA, e foram submetidas ao teste de normalidade de Shapiro-Wilk para definir o teste estatístico (p<0,05). Na análise histológica, o grupo CLA apresentou predominantemente necrose no terço oclusal e médio da polpa coronária, enquanto o BS-CLA apresentou inflamação severa no terço oclusal e moderada no terço médio, ambos diferentes do Controle (p<0,05). O grupo CLA+BS apresentou inflamação moderada no terço oclusal, sem diferença para o grupo BS-CLA (p>0,05), inflamação leve no terço médio, e ausência de inflamação no terço cervical sem diferença para o Controle (p>0,05). O grupo CLA+ H2O apresentou inflamação leve no terço oclusal e ausente nos demais terços, sem diferença para o grupo CLA+BS e Controle (p>0,05). No terço cervical, o grupo CLA apresentou inflamação moderada, diferente do grupo Controle (p<0,05). Os grupos CLA e BS-CLA, apresentaram maior penetração de H2O2 (p<0,05). Na alteração de cor imediata e após 24 horas, os grupos CLA, BS-CLA e CLA+BS, apresentaram alteração de cor semelhante (p>0,05). Conclusão: O Biosilicato® é capaz de reduzir o processo inflamatório causado pelo gel clareador e auxilia no processo de reparo. O protocolo de aplicação do BS misturado com o gel clareador apresentou melhores resultados, uma vez que reduz a penetração do gel clareador e mantém a eficácia clareadora.
New products are being developed with the purpose of mitigating the effects of the compounds used in bleaching procedures. The objective of the present study was to evaluate in vivo the therapeutic potential of Biosilicate® (BS) gel on the pulp tissue of Wistar rats by means of histological and immunohistochemical analysis, as well as to analyze in vitro, the penetration of H2O2 and bleaching efficiency in bovine teeth. For this, this study was divided into 2 parts. Part 1: In vivo segment: 50 hemi-maxilla of 25 rats were randomly divided into 5 groups (n = 10): Control- received no treatment; CLA- received 30 min of the 35% H2O2 bleaching gel; BS-CLA- received 20 min of BS gel, followed by 30 min of 35% H2O2; CLA+BS- received 30 min of a mixture of 35% H2O2 with the BS gel (1:1); CLA+H2O- received 30 min of a mixture of 35% H2O2 with distilled water (1:1). After 2 days, the animals were killed and the hemi-maxils separated for histological analysis in H.E. Inflammation scores and data submitted to the Kruskal-Wallis and Dunn tests and Mann-Whitney test (p<0.05) were assigned. Segment in vitro: 50 discs of bovine teeth were coupled in artificial pulp chambers, and divided into 5 groups (n=10) that received the same treatments as the in vivo segment. The penetration of H2O2 by enamel and dentin was quantified based on the reaction of this with the leucocrystal violet dye. The color change was analyzed by the CIELab system, in the periods: before the bleaching session - (T0), immediately after (T1), and 24 hours after (T2), and data submitted to analysis of variance (ANOVA) followed by the test Tukey (p<0.05). Part 2: 40 hemi-maxillae of 20 rats were randomly divided into 4 groups (n=10): Control- received no treatment; CLA- received 30 min of the 35% H2O2 bleaching gel; BS-CLA- received 20 min of BS gel, followed by 30 min of 35% H2O2; CLA+BS- received 30 min of a mixture of 35% H2O2 with the BS gel (1:1). After 2 days, the animals were killed and the hemi-maxilla separated for histological analysis in HE and immunohistochemistry for PCNA, HO-1, TNF-α and P-Substance. Scores were assigned to TNF-α inflammation and immunostaining, and submitted to the Kruskal-Wallis and Dunn tests (p<0.05). For the analysis of the area corresponding to the immunostaining obtained through the optical density of the Substance P, an analysis of the variance (ANOVA) was performed, followed by the Tukey test for comparison between the means. We counted the immunolabellated cells for HO-1 and PCNA and were submitted to the Shapiro-Wilk normality test to define the statistical test (p<0.05). In the histological analysis, the CLA group presented predominantly necrosis in the occlusal and middle third of the coronary pulp, whereas the BS-CLA presented severe inflammation in the occlusal third and moderate in the middle third, both different from the Control (p<0.05). The CLA+BS group presented moderate inflammation in the occlusal third, with no difference for the BS-CLA group (p> 0.05), slight inflammation in the middle third, and absence of inflammation in the cervical third without difference for Control (p>0,05). The CLA+H2O group had mild inflammation in the occlusal third and absent in the remaining thirds, with no difference for CLA+BS and Control group (p>0.05). In the cervical third, the CLA group had moderate inflammation, different from the Control group (p<0.05). The CLA and BS-CLA groups presented higher penetration of H2O2 (p<0.05). At the immediate color change and after 24 hours, the CLA, BS-CLA and CLA+BS groups presented similar color changes (p>0.05).Conclusion: Given the results obtained in both studies, it can be concluded that Biosilicate® is able to reduce the inflammatory process caused by the bleaching gel and helps in the repair process. The application protocol of BS blended with the bleaching gel showed better results as it reduces the penetration of the bleaching gel and maintains bleaching efficacy.
New products are being developed with the purpose of mitigating the effects of the compounds used in bleaching procedures. The objective of the present study was to evaluate in vivo the therapeutic potential of Biosilicate® (BS) gel on the pulp tissue of Wistar rats by means of histological and immunohistochemical analysis, as well as to analyze in vitro, the penetration of H2O2 and bleaching efficiency in bovine teeth. For this, this study was divided into 2 parts. Part 1: In vivo segment: 50 hemi-maxilla of 25 rats were randomly divided into 5 groups (n = 10): Control- received no treatment; CLA- received 30 min of the 35% H2O2 bleaching gel; BS-CLA- received 20 min of BS gel, followed by 30 min of 35% H2O2; CLA+BS- received 30 min of a mixture of 35% H2O2 with the BS gel (1:1); CLA+H2O- received 30 min of a mixture of 35% H2O2 with distilled water (1:1). After 2 days, the animals were killed and the hemi-maxils separated for histological analysis in H.E. Inflammation scores and data submitted to the Kruskal-Wallis and Dunn tests and Mann-Whitney test (p<0.05) were assigned. Segment in vitro: 50 discs of bovine teeth were coupled in artificial pulp chambers, and divided into 5 groups (n=10) that received the same treatments as the in vivo segment. The penetration of H2O2 by enamel and dentin was quantified based on the reaction of this with the leucocrystal violet dye. The color change was analyzed by the CIELab system, in the periods: before the bleaching session - (T0), immediately after (T1), and 24 hours after (T2), and data submitted to analysis of variance (ANOVA) followed by the test Tukey (p<0.05). Part 2: 40 hemi-maxillae of 20 rats were randomly divided into 4 groups (n=10): Control- received no treatment; CLA- received 30 min of the 35% H2O2 bleaching gel; BS-CLA- received 20 min of BS gel, followed by 30 min of 35% H2O2; CLA+BS- received 30 min of a mixture of 35% H2O2 with the BS gel (1:1). After 2 days, the animals were killed and the hemi-maxilla separated for histological analysis in HE and immunohistochemistry for PCNA, HO-1, TNF-α and P-Substance. Scores were assigned to TNF-α inflammation and immunostaining, and submitted to the Kruskal-Wallis and Dunn tests (p<0.05). For the analysis of the area corresponding to the immunostaining obtained through the optical density of the Substance P, an analysis of the variance (ANOVA) was performed, followed by the Tukey test for comparison between the means. We counted the immunolabellated cells for HO-1 and PCNA and were submitted to the Shapiro-Wilk normality test to define the statistical test (p<0.05). In the histological analysis, the CLA group presented predominantly necrosis in the occlusal and middle third of the coronary pulp, whereas the BS-CLA presented severe inflammation in the occlusal third and moderate in the middle third, both different from the Control (p<0.05). The CLA+BS group presented moderate inflammation in the occlusal third, with no difference for the BS-CLA group (p> 0.05), slight inflammation in the middle third, and absence of inflammation in the cervical third without difference for Control (p>0,05). The CLA+H2O group had mild inflammation in the occlusal third and absent in the remaining thirds, with no difference for CLA+BS and Control group (p>0.05). In the cervical third, the CLA group had moderate inflammation, different from the Control group (p<0.05). The CLA and BS-CLA groups presented higher penetration of H2O2 (p<0.05). At the immediate color change and after 24 hours, the CLA, BS-CLA and CLA+BS groups presented similar color changes (p>0.05).Conclusion: Given the results obtained in both studies, it can be concluded that Biosilicate® is able to reduce the inflammatory process caused by the bleaching gel and helps in the repair process. The application protocol of BS blended with the bleaching gel showed better results as it reduces the penetration of the bleaching gel and maintains bleaching efficacy.
Descrição
Palavras-chave
Clareamento dental, Peróxido de hidrogênio, Pulpite, Tooth bleaching, Hydrogen peroxide, Pulpitis