DBPapers
DOI: 10.5593/SGEM2014/B61/S25.044

COMPARATION BETWEEN TWO IN VITRO TESTS FOR TOXICITY ASSESSING OF A LIPOPHYLE EXTRACT OF PELLOIDIC MUD FROM TECHIRGHIOL LAKE, CONSTANTA, ROMANIA

E. Doroftei, L. Moldovan, O. Craciunescu, T. Negreanu-Pirjol, H. Miresan, M. M. Bratu
Wednesday 1 October 2014 by Libadmin2014

References: 14th International Multidisciplinary Scientific GeoConference SGEM 2014, www.sgem.org, SGEM2014 Conference Proceedings, ISBN 978-619-7105-20-9 / ISSN 1314-2704, June 19-25, 2014, Book 6, Vol. 1, 313-320 pp

ABSTRACT
The aim of this paper was to assess cyto- and genotoxic effects of a lipophyle extract of pelloidic mud (LE) from Techirghiol Lake, Romania, using two in vitro tests: cell viability determination in a mice fibroblasts culture (NCTC, clone L 929) and Triticum assay. The extract contains a mixture of saturated and unsaturated hidrocarbonated compounds with carbonilic and carboxylic groups. Cytotoxic effect of LE, in a dilution ranging between 0.025- 1 g/mL, on the mice fibroblasts culture was assessed by neutral red (NR) test after 24 and 48 h of cultivation according to ISO 10993-5:2003. Cell viability was calculated by comparison with the cell culture control considered to be 100% viable cells, at each period of time. The morphological observations were done on culture plates after the cultured cells were fixed in methanol and Giemsa stained. In Triticum assay we used two concentrations of LE in alcoholic solutions (0.025 g/mL and 0.045 g/mL). The Triticum seeds were preliminary imbued in water, and then they were treated for 6, 12 and 24 hours in LE solutions. The control group was treated with tap water. In the end of the experiment the harvested root tips were prepared according to Feulgen’s squash technique using Schiff reagent. The cytotoxic effects of LE were investigated by calculating the mitotic index and also through the analysis of chromosomes alterations during the mitosis. In mice fibroblasts culture the LE expressed a cytotoxic effect only in higher amounts than 0.066 g/mL. In lower concentrations of LE, the cells had 95% viability and there were no morphological changes, after 48 h of cultivation. Triticum assay showed that the powerful effect on the cell division was in 0.025 g/mL LE after 12 or 24 hours of treatment. It was observed a rise of mitotic index and a small number of chromosomal aberrations in all variants. Because the studied concentrations of LE on the seeds of Triticum aestivum were smaller than 0.066 g/mL, we can notice that LE solutions had cellular regeneration effects and also cell metabolism stimulating. As conclusions: LE was not cytotoxic in the range of concentration 0-0.066 g/mL and can be used in cosmetic products. The information given from the two assays correlated each other and offered a better image concerning the toxic effects of LE. The two methods could be used together in the future studies as alternative methods for toxicity assay.

Keywords: pelloidic mud, fibroblasts culture, Triticum assay, cytotoxic effect