*2.4. Cytotoxicity Assays*

The exposure concentrations were prepared in medium, with the highest concentration being 100 μg/mL. For the control group culture medium without the extract was used. Moreover, a control of solvent (0.5% of DMSO) has also been included. The exposure concentrations were added to both cells and the cytotoxicity assays were performed after 24 and 48 h of exposure.

Neutral red uptake (NR) was evaluated according to Borenfreund and Puerner (1984) [26] with modifications [27]. MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)- 2-(4-sulfophenyl)- 2H- tetrazolium salt) reduction was measured according to the procedure of Baltrop et al. (1991) [28]. Moreover, total protein content (TP) was performed according to the procedure described by Bradford (1976) [29].
