Influência da espessura do substrato dental sobre a eficácia clareadora e citotoxicidade de diferentes protocolos de clareamento
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Data
2016-03-23
Autores
Duque, Carla Caroline de Oliveira [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Tem sido demonstrado que a susceptibilidade do complexo dentino-pulpar aos efeitos adversos do clareamento dental apresenta uma relação direta com a espessura de esmalte/dentina do substrato dental. Desta forma, o objetivo do presente estudo foi avaliar o efeito estético e biológico de um gel clareador contendo 10% de peróxido de hidrogênio (H2O2) aplicado por diferentes períodos sobre a superfície de discos simulando a espessura de incisivos inferiores (ICI) e pré-molares superiores (PMS). Discos de esmalte/dentina, provenientes de incisivos bovinos, com 2,3 e 4,0 mm de espessura foram adaptados a câmaras pulpares artificiais e distribuídos nos Grupos ICI e PMS, respectivamente. O gel clareador com 10% de H2O2 foi aplicado na superfície de esmalte por 3x15, 1x15 ou 1x5 minutos. Como controle positivo, um gel com 35% de H2O2 aplicado por 3x15 minutos (protocolo tradicional) foi empregado. Nenhum tratamento foi realizado na superfície de esmalte no grupo controle negativo. O meio de cultura em contato com a dentina imediatamente após o clareamento (extrato) foi coletado e aplicado por 1 hora sobre células pulpares humanas. A viabilidade e morfologia celular foram avaliados imediatamente após exposição aos extratos (T1), bem como 72 horas pós-tratamento (T2). O estresse oxidativo e a quantificação do H2O2 nos extratos também foram analisados em T1. A atividade de ALP (atividade de fosfatase alcalina) e deposição de nódulos de mineralização (NM) foram avaliados em períodos de 14 e 21 dias pós-clareamento, respectivamente. De forma a avaliar a eficácia estética dos protocolos testados nas diferentes condições experimentais, mensurou-se ainda a alteração de cor (ΔE) durante seis sessões clareadoras. De uma forma geral, observou-se que todos os protocolos experimentais com gel a 10% de H2O2 promoveram diminuição dos efeitos deletérios promovidos pelo protocolo tradicional. O protocolo 10% 3x15 causou redução significativa na viabilidade celular em relação ao controle negativo no período T1, em ambos os Grupos ICI e PMS, sendo que as células expostas a este protocolo no Grupo ICI não demonstraram capacidade proliferativa em T2. As alterações morfológicas, a intensidade do estresse oxidativo e a redução nos marcadores fenotípicos (ALP e NM) também foram mais intensos para este protocolo nos Grupos ICI e PMS. Para os protocolos 10% 1x15 ou 1x5 minutos, ausência de redução significante na viabilidade celular foi observada, independente da espessura. No entanto, estresse oxidativo significativo foi observado nas células do Grupo ICI. Estes protocolos promoveram redução na atividade de ALP, a qual foi proporcional ao tempo de contato e espessura do substrato dental; entretanto não se observou redução na deposição de NM aos 21 dias. Valores de ΔE similares ao protocolo tradicional foram observados para os protocolos 10% 3x15 e 10% 1x15 no Grupo ICI após 4 e 6 sessões, respectivamente. Para o Grupo PMS, este parâmetro foi atingido apenas para o protocolo 10% 3x15 após 6 sessões clareadoras. Concluiu-se que a espessura do substrato dental influenciou significativamente na citotoxicidade trans-amelodentinária do gel com 10% de H2O2. A aplicação deste gel por períodos de 45 ou 15 minutos sobre discos simulando a espessura de incisivos inferiores minimizou significativamente a toxicidade celular e promoveu clareamento tão efetivo quanto o protocolo tradicionalmente empregado.
The susceptibility of pulp-dentin complex to the adverse effects of tooth bleaching therapies has a direct relationship with the thickness of dental substrate. Therefore, the aim of this study was to assess the biologic and esthetic effect of a 10% hydrogen peroxide (H2O2) bleaching gel applied for different periods onto enamel/dentin discs simulating the thickness of low incisors (LI) and upper pre-molars (PM). Enamel/dentin discs from bovine incisors measuring 2.3 and 4.0 mm were adapted to artificial pulp chambers and distributed into the IC Group (low incisors) and PM Group (upper premolars), respectively. The enamel surface was bleached with a 10% H2O2 gel for 3x15, 1x15 or 1x5 minutes. The 35% H2O2 gel applied for 3x15 minutes was used as positive control (traditional therapy) and no treatment was performed on negative control group. The culture medium immediately after bleaching (extract) was applied for 1 hour on human dental pulp cells. Cell viability and morphology were evaluated immediately after bleaching (T1) and 72 h thereafter (T2). Oxidative stress and the amount of H2O2 in culture medium were also quantified at T1. The ALP activity and mineralized nodule (MN) deposition were assessed 14 and 21 days after bleaching, respectively. The color change (ΔE) of enamel was analyzed throughout six bleaching sessions. Overall, the experimental protocols with the 10% H2O2 gel minimized significantly the negative effects of traditional therapy to cultured pulp cells. The protocol 10% 3x15 promoted significant cell viability reduction in comparison with negative control at T1, in both, IC and PM Groups; however, the cells of IC Group did not feature proliferative capability at T2. The morphologic alterations, oxidative stress intensity and reduction on phenotype markers (ALP and NM) expression were also more intense on IC Group for this bleaching protocol. Regarding the protocols 10% 1x15 and 1x5, no significant cell viability reduction related to negative control was observed regardless of the thickness of dental substrate. Nevertheless, significant oxidative stress was observed on cells from IC Group. These protocols caused significant reduction of ALP activity in comparison to negative control, which intensity being proportional to enamel/dentin thickness; however, no reduction on NM was detected at 21 days post-bleaching. Similar ΔE values as positive control were found for the 10% 3x15 and 1x15 protocol on IC Group after 4 and 6 sessions, respectively. For PM Group, this feature was reached out after 6 bleaching sessions for the 10% 3x15 protocol. It was concluded that the enamel/dentin thickness of dental substrate played a significant role on the trans-enamel and trans-dentinal cytotoxicity of a 10% H2O2 bleaching gel on human dental pulp cells in vitro. Application of gel during 45 or 15 minutes onto thin dental substrate minimizes significantly the cell toxicity in comparison to high concentrated gels (35%) associated with similar esthetic outcome by increasing the number of bleaching sessions.
The susceptibility of pulp-dentin complex to the adverse effects of tooth bleaching therapies has a direct relationship with the thickness of dental substrate. Therefore, the aim of this study was to assess the biologic and esthetic effect of a 10% hydrogen peroxide (H2O2) bleaching gel applied for different periods onto enamel/dentin discs simulating the thickness of low incisors (LI) and upper pre-molars (PM). Enamel/dentin discs from bovine incisors measuring 2.3 and 4.0 mm were adapted to artificial pulp chambers and distributed into the IC Group (low incisors) and PM Group (upper premolars), respectively. The enamel surface was bleached with a 10% H2O2 gel for 3x15, 1x15 or 1x5 minutes. The 35% H2O2 gel applied for 3x15 minutes was used as positive control (traditional therapy) and no treatment was performed on negative control group. The culture medium immediately after bleaching (extract) was applied for 1 hour on human dental pulp cells. Cell viability and morphology were evaluated immediately after bleaching (T1) and 72 h thereafter (T2). Oxidative stress and the amount of H2O2 in culture medium were also quantified at T1. The ALP activity and mineralized nodule (MN) deposition were assessed 14 and 21 days after bleaching, respectively. The color change (ΔE) of enamel was analyzed throughout six bleaching sessions. Overall, the experimental protocols with the 10% H2O2 gel minimized significantly the negative effects of traditional therapy to cultured pulp cells. The protocol 10% 3x15 promoted significant cell viability reduction in comparison with negative control at T1, in both, IC and PM Groups; however, the cells of IC Group did not feature proliferative capability at T2. The morphologic alterations, oxidative stress intensity and reduction on phenotype markers (ALP and NM) expression were also more intense on IC Group for this bleaching protocol. Regarding the protocols 10% 1x15 and 1x5, no significant cell viability reduction related to negative control was observed regardless of the thickness of dental substrate. Nevertheless, significant oxidative stress was observed on cells from IC Group. These protocols caused significant reduction of ALP activity in comparison to negative control, which intensity being proportional to enamel/dentin thickness; however, no reduction on NM was detected at 21 days post-bleaching. Similar ΔE values as positive control were found for the 10% 3x15 and 1x15 protocol on IC Group after 4 and 6 sessions, respectively. For PM Group, this feature was reached out after 6 bleaching sessions for the 10% 3x15 protocol. It was concluded that the enamel/dentin thickness of dental substrate played a significant role on the trans-enamel and trans-dentinal cytotoxicity of a 10% H2O2 bleaching gel on human dental pulp cells in vitro. Application of gel during 45 or 15 minutes onto thin dental substrate minimizes significantly the cell toxicity in comparison to high concentrated gels (35%) associated with similar esthetic outcome by increasing the number of bleaching sessions.
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Palavras-chave
Citotoxicidade, Clareamento dental, Polpa dental, Cytotoxicity, Tooth-bleaching, Dental pulp