Citotoxicidade e capacidade de limpeza de soluções com potencial para uso em endodontia
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Data
2018-03-21
Autores
Coaguila Llerena, Eric Hernán [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O hipoclorito de cálcio [Ca(OCl)2] e o cloridrato de octenidina (OCT) têm sido estudados como potenciais soluções irrigadoras endodônticas alternativas ao hipoclorito de sódio (NaOCl) ou à clorexidina (CHX). Os objetivos deste estudo foram avaliar a citotoxicidade do Ca(OCl)2 e do OCT (Publicação 1), e o efeito do OCT na limpeza do canal radicular (Publicação 2). Para a avaliação da citotoxicidade, foram utilizadas células do ligamento periodontal humano (hPDL) e fibroblastos da linhagem L929. As células foram expostas a diferentes diluições das soluções de Ca(OCl)2 a 2,5 e 5%, OCT a 0,1%, NaOCl a 2,5% e CHX a 2%. A viabilidade celular foi avaliada por meio dos ensaios do metil-tiazol-tetrazólio (MTT) e do vermelho neutro (NR), e a migração celular pelo teste de cicatrização. A análise estatística foi efetuada empregando ANOVA de dois fatores e teste de Bonferroni (α=0,05). Os ensaios MTT e NR mostraram, nas células do hPDL, que o OCT a 0,1% foi menos citotóxico (P<0,05), seguido da CHX a 2% e do Ca(OCl)2 a 2,5% (P<0,05). Não houve diferença significativa entre NaOCl a 2,5% e Ca(OCl)2 a 5% (P>0,05), sendo que estas soluções apresentaram maior citotoxicidade que as demais. Em L929, o resultado foi similar, exceto que não houve diferença significativa entre CHX a 2% e Ca(OCl)2 a 2,5% (P>0,05). No teste de cicatrização, tanto nas células do hPDL quanto L929, a migração celular do OCT a 0,1%, CHX a 2%, Ca(OCl)2 a 2,5% foi significativamente maior do que o Ca(OCl)2 a 5% e NaOCl a 2,5% às 24 horas (P<0,05). Para a avaliação da capacidade de limpeza do OCT, foram utilizados 50 dentes unirradiculados humanos extraídos que foram distribuídos aleatoriamente em 5 grupos (n=10) de acordo com as soluções irrigadoras utilizadas no preparo biomecânico do canal radicular: G1, OCT a 0,1%; G2, CHX a 2%; G3, NaOCl a 2,5%; G4, OCT + ácido etilenodiaminotetracético (EDTA) a 17% e G5, NaOCl a 2,5% + EDTA a 17%. A smear layer foi avaliada em microscópio eletrônico de varredura utilizando sistema de escores. Os resultados foram analisados pelos testes de Kruskal-Wallis e Dunn (α = 0,05). Não houve diferença significativa entre os grupos OCT, CHX e NaOCl (P>0,05), e esses grupos apresentaram maiores valores de smear layer que os grupos NaOCl + EDTA e OCT + EDTA (P<0,05). Não houve diferença significativa entre os grupos NaOCl + EDTA e OCT + EDTA (P>0,05). Pode-se concluir que, nas linhagens celulares estudadas, o OCT a 0,1% foi a solução menos citotóxica, e que o Ca(OCl)2 a 2,5% e a 5% apresentaram citotoxicidade menor ou similar ao NaOCl a 2,5%, respectivamente. O OCT, utilizado como solução irrigadora única, não apresentou capacidade de limpeza e deveria ser usado em associação com EDTA como irrigante final para se obter remoção da smear layer.
Calcium hypochlorite [Ca(OCl)2] and octenidine hydrochloride (OCT) have been studied as alternative irrigating solutions to sodium hypochlorite (NaOCl) or chlorhexidine (CHX). The objectives of this study were to evaluate the cytotoxicity of Ca(OCl)2 and OCT (Publication 1), and the effect of OCT on root canal cleaning (Publication 2). For cytotoxicity evaluation, human periodontal ligament (hPDL) cells and permanent cell line of mouse fibroblasts L929 were used. The cells were exposed to different doses of different solutions: 2.5% and 5% Ca(OCl)2, 0.1% OCT, 2.5% NaOCl and 2% CHX for 10 minutes. Cell viability was assessed by methyl-thiazoltetrazolium (MTT) and neutral red (NR) assays, and cell migration was determined by the scratch assay. Statistical analysis was performed by two-way ANOVA and Bonferroni tests (α=0.05). The MTT and NR assays revealed that 0.1% OCT was less cytotoxic in hPDL cells (P<0.05), followed by 2% CHX and 2.5% Ca(OCl)2 (P<0.05). There was no significant difference between 2.5% NaOCl and 5% Ca(OCl)2 (P>0.05), but these solutions showed greater cytotoxicity than the others. The result was the same for L929 cells, except that there was no significant difference between 2% CHX and 2.5% Ca(OCl)2 (P>0.05). Scratch assay in L929 and hPDL cells showed that cell migration of 0.1% OCT, 2% CHX and 2.5% Ca(OCl)2 groups was higher than 5% Ca(OCl)2 and 2.5% NaOCl groups at 24 hours (P<0.05). For evaluation of OCT cleaning capacity, fifty human unirradicular extracted teeth were randomly distributed in 5 groups (n=10) according to irrigant solutions which were used during root canal preparation: G1, 0.1% OCT; G2, 2% chlorhexidine (CHX); G3, 2.5% sodium hypochlorite (NaOCl); G4, OCT + 17% ethylenediaminetetraacetic acid (EDTA) and G5, 2.5% NaOCl + 17%. The smear layer was evaluated using a score system and the data were analyzed by Kruskal-Wallis and Dunn (α=0.05). In all root canal thirds there was no significant difference between OCT, CHX and NaOCl groups (P>0.05), and these groups showed higher smear layer values than NaOCl + EDTA and OCT + EDTA groups (P<0.05). There was no significant difference between NaOCl + EDTA and OCT + EDTA groups (P>0.05). In conclusion, in tested cell lines, 0.1% OCT had lower cytotoxicity than CHX, Ca(OCl)2 and NaOCl. Ca(OCl)2 at concentrations of 2.5% and 5% showed lower or similar cytotoxicity to 2.5% NaOCl, respectively. The OCT used as a single root canal irrigant presented poor cleaning capacity and could be used in association with a final irrigation with EDTA to obtain smear layer removal.
Calcium hypochlorite [Ca(OCl)2] and octenidine hydrochloride (OCT) have been studied as alternative irrigating solutions to sodium hypochlorite (NaOCl) or chlorhexidine (CHX). The objectives of this study were to evaluate the cytotoxicity of Ca(OCl)2 and OCT (Publication 1), and the effect of OCT on root canal cleaning (Publication 2). For cytotoxicity evaluation, human periodontal ligament (hPDL) cells and permanent cell line of mouse fibroblasts L929 were used. The cells were exposed to different doses of different solutions: 2.5% and 5% Ca(OCl)2, 0.1% OCT, 2.5% NaOCl and 2% CHX for 10 minutes. Cell viability was assessed by methyl-thiazoltetrazolium (MTT) and neutral red (NR) assays, and cell migration was determined by the scratch assay. Statistical analysis was performed by two-way ANOVA and Bonferroni tests (α=0.05). The MTT and NR assays revealed that 0.1% OCT was less cytotoxic in hPDL cells (P<0.05), followed by 2% CHX and 2.5% Ca(OCl)2 (P<0.05). There was no significant difference between 2.5% NaOCl and 5% Ca(OCl)2 (P>0.05), but these solutions showed greater cytotoxicity than the others. The result was the same for L929 cells, except that there was no significant difference between 2% CHX and 2.5% Ca(OCl)2 (P>0.05). Scratch assay in L929 and hPDL cells showed that cell migration of 0.1% OCT, 2% CHX and 2.5% Ca(OCl)2 groups was higher than 5% Ca(OCl)2 and 2.5% NaOCl groups at 24 hours (P<0.05). For evaluation of OCT cleaning capacity, fifty human unirradicular extracted teeth were randomly distributed in 5 groups (n=10) according to irrigant solutions which were used during root canal preparation: G1, 0.1% OCT; G2, 2% chlorhexidine (CHX); G3, 2.5% sodium hypochlorite (NaOCl); G4, OCT + 17% ethylenediaminetetraacetic acid (EDTA) and G5, 2.5% NaOCl + 17%. The smear layer was evaluated using a score system and the data were analyzed by Kruskal-Wallis and Dunn (α=0.05). In all root canal thirds there was no significant difference between OCT, CHX and NaOCl groups (P>0.05), and these groups showed higher smear layer values than NaOCl + EDTA and OCT + EDTA groups (P<0.05). There was no significant difference between NaOCl + EDTA and OCT + EDTA groups (P>0.05). In conclusion, in tested cell lines, 0.1% OCT had lower cytotoxicity than CHX, Ca(OCl)2 and NaOCl. Ca(OCl)2 at concentrations of 2.5% and 5% showed lower or similar cytotoxicity to 2.5% NaOCl, respectively. The OCT used as a single root canal irrigant presented poor cleaning capacity and could be used in association with a final irrigation with EDTA to obtain smear layer removal.
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Palavras-chave
Endodontia, Microscopia eletrônica de varredura, Teste de materiais, Tratamento do canal radicular, Endodontics, Scanning electron microscopy, Materials testing, Root canal therapy