In vitro genotoxicity of root canal sealers.

AIM: To evaluate the effect of leakage on differences in genotoxicity of root canal sealers ex vivo according to their main components using two different cytogenetic assays. METHODOLOGY: Six materials of different composition (GuttaFlow, Epiphany, Diaket, IRM, SuperEBA and Hermetic) were tested on human peripheral blood lymphocytes using the comet assay and chromosomal aberration analysis. Prepared materials were eluted in physiological solution for 1 h, 1 day, 5 and 30 days. Thereafter cultures were treated with 8 microg, 4 microg and 2 microg of each sealer. Frequencies of chromatide and chromosome breaks and accentric fragments were determined. Comet assay was used to evaluate primary DNA damage by measuring tail length and tail intensity. Chi-square, Fisher's PLSD (Protected Least Significant Difference) and Kruskall-Wallis non parametric tests were used for statistical analysis. RESULTS: After 1-h elution only the highest dose of Diaket, Hermetic and SuperEBA significantly (P = 0.035, P = 0.048, P = 0.037 respectively) affected the measured cytogenetic parameters. The migration ability of DNA was more strongly affected than induction of chromosomal aberrations. After elutions longer than 24 h none of the tested sealers exhibited a genotoxic effect. CONCLUSION: Under the conditions used in the study all sealers had acceptable biocompatibility in terms of genotoxicity.

Chicken nucleated blood cells as a cellular model for genotoxicity testing using the comet assay.

Mycotoxins can frequently occur in animal feed and human food. T-2 toxin, as the most toxic trichothecene, has been implicated as the causative agent in a variety of animal diseases and is associated with some human diseases. The comet assay was performed as a test for detection of DNA damage caused by T-2 toxin in peripheral blood cells of chicken. The suitability of the comet assay as a biomarker for genotoxic analysis has been applied in studies using human white blood cells. It can be applied to any tissue from which a single cell suspension can be obtained. The method has already been applied to chicken as a foodstuff for detection of irradiation of food containing DNA. However, application of the method on chicken blood cells has not been set up yet. The aim of this research was to develop a protocol for detection of DNA damage induced by T-2 toxin in chicken blood cells. Chickens were administered orally with T-2 toxin and the samples of whole blood were collected at 24 h post treatment. The DNA damage was determined by an increase in the comet parameters in tested animals. Our results show that T-2 toxin had induced significant DNA damage in treated chicken as compared with control animals, indicating that the assay can be used for the assessment of primary DNA damage caused by mycotoxins.