Cytotoxicity analysis of EDTA and citric acid applied on murine resident macrophages culture.
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.
Advisor:Giulio Gavini; Antonio Carlos Bombana; Primavera Borelli Garcia; Giulio Gavini
School:Universidade de São Paulo
Source Type:Master's Thesis
Keywords:Citric acid EDTA Macrophages Root canal irrigants Toxicity Toxicity-cell culture
Date of Publication:12/08/2004