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Master's Dissertation
DOI
10.11606/D.23.2004.tde-15032005-122159
Document
Author
Full name
Kali Fatima Amaral
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2004
Supervisor
Committee
Gavini, Giulio (President)
Bombana, Antonio Carlos
Garcia, Primavera Borelli
Title in Portuguese
"Análise da citotoxicidade do EDTA e do ácido cítrico aplicados em cultura de macrófagos peritoneais residentes"
Keywords in Portuguese
Ácido cítrico
EDTA
Irrigantes do canal radicular
Macrófagos
Toxicidade
Toxicidade–cultura de células
Abstract in Portuguese
O presente estudo avaliou in vitro o efeito citotóxico das soluções de EDTA a 17% e ácido cítrico a 15% sob macrófagos peritoneais residentes, valendo-se do método MTT. Após anestesia e sacrifício de 32 camundongos Swiss machos, procedeu-se a coleta do exsudato celular pela injeção e aspiração de meio de cultura estéril na cavidade abdominal dos animais. Pelo processamento do exsudato peritoneal, obteve-se em média 95% de macrófagos. Alíquotas de 5 x 10 5 células foram plaqueadas em triplicata, de acordo com os grupos experimentais. Diluições de 0,5% de EDTA e ácido cítrico foram adicionadas ao meio de cultura num total de 1 mL. O grupo controle recebeu somente meio de cultura estéril. Verificou-se a citotoxicidade em dois momentos: períodos de curto prazo (0, 6, 12 e 24 horas) e médio prazo (1,3, 5, 7 dias). Ao final dos referidos tempos de observação, as amostras foram tratadas pelo corante MTT, obtendo-se valores de absorbância em leitora ELISA 550 nm. Todo o procedimento experimental foi repetido 2 vezes. No período de curto prazo, a análise de variância apontou diferenças significantes (p< 0,05), sendo Fc=46,07 contra Ft= 3,15, para os grupos avaliados: os Grupos EDTA (0,253 nm) e ácido cítrico (0,260 nm) foram mais citotóxicos que o Grupo controle (0,355 nm). Observações de médio prazo revelaram significância estatística (p< 0,05) entre os grupos, sendo Fc= 171,0 contra Ft= 3,15. Ambas as soluções, EDTA (0,158 nm) e ácido cítrico (0,219 nm), mostraram maior toxicidade em relação ao controle (0,310nm), porém o EDTA apresentou-se mais citotóxico que o ácido cítrico, reduzindo substancialmente a população macrofágica. Como conclusão, as soluções irrigantes testadas exerceram toxicidade aos macrófagos peritoneais em cultura, no entanto, a viabilidade tardia destas células foi menos alterada pelo ácido cítrico
Title in English
Cytotoxicity analysis of EDTA and citric acid applied on murine resident macrophages culture.
Keywords in English
Citric acid
EDTA
Macrophages
Root canal irrigants
Toxicity
Toxicity-cell culture
Abstract in English
The present study evaluated in vitro cytotoxic effects of 17% EDTA and 15% citric acid in murine resident macrophages using MTT assay. After anesthesia and sacrifice of thirty two Swiss male mice, it had processed the peritoneal cellular exudates by fresh culture medium injection and aspiration in peritoneal animals cavities. The peritoneal exudates were composed approximately by 95% of macrophages. 5x 10 5 cells were plated in triplicate, according experimental groups. Each 0.5% dilutions of EDTA and citric acid were applied in medium culture, resulting 1 mL volume. Fresh medium served as control. The cytotoxicity was evaluated in two moments: short term (0, 6, 12, 24 hours) and long term (1, 3, 5, 7 days). After tested periods, the samples were treated by MTT ink assay and absorbance was determined using ELISA microplate reader at 550 nm. All these procedures were repeated twice. To short term period, ANOVA showed significant differences (p< 0.05) among groups (Fc= 46.07 x Ft= 3.15). EDTA (0.253 nm) and citric acid (0.260 nm) groups exhibited more cytotoxicity than control group. Long term observations exhibited statistical differences (p< 0.05), that Fc= 171.0 x Ft= 3.15. EDTA (0.158 nm) and citric acid (0.219 nm) solutions were cytotoxic when compared to control group, thus EDTA reduced greater macrophages viability than citric acid. Based on our results it seems that final irrigants tested presented toxic effects to murine macrophages culture, but in long term evaluation citric acid was considered less irritant.
 
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Publishing Date
2005-04-14
 
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