Ensaio de absorção de vermelho neutro

O ensaio de absorção de vermelho neutro (VN), (cloridrato de 3- amino-7-

dimetilamino-2-metilfenazina) um corante vital solúvel em água, em passar pela
membrana plasmática e tornar-se concentrado nos lisossomos de células viáveis
(TRIGLIA et al., 1991). Initially the test was applied to determine which concentrations
would be tested, based on the literature, which reported that the most studied
concentrations for Ag4V2O7:β AgVO3 vary from 7,8 to 125 µg/ml. The concentrations
of (125, 62,5, 31,25, 15,6 and 7,8µg/mL) were chosen to carry out the study. In a 96-
well plate, 1x104 cells/well were seeded and exposed to Ag4V2O7:β AgVO3. Após o
tratamento, o meio de cultura foi retirado, adicionou-se 200 µL de solução de vermelho
neutro (30 µg/mL) e as células foram incubadas por 2 horas. Após esse tempo, as
células foram lavadas com PBS 1X e elui-se o VN com 200 µL da solução de etanol
50% (v/v) e ácido acético 1% (v/v) seguido de incubação por 10min em temperatura
ambiente. A absorbância foi medida em 540 nm. Todos os testes foram conduzidos em
triplicata. O cálculo utilizado para apresentação dos resultados foi:

Experimental group
% cytotoxicity= x 100
Control groupmean

Neutral red absorption assay

The neutral red absorption assay (NR), (3- amino-dimethylamino-2-methylphenazine) is

a vital water-soluble dye, passing through the plasma membrane and becoming
concentrated in the lysosomes of viable cells (TRIGLIA et al., 1991). Initially the test
was applied to determine which concentrations would be tested, based on the literature,
which reported that the most studied concentrations for Ag4V2O7:β AgVO3 vary from
7.8 to 125 μg/ml. The concentrations of (125, 62.5, 31.25, 15.6 and 7.8μg/mL) were
chosen to carry out the study. In a 96-well plate, 1x104 cells/well were seeded and
exposed to Ag4V2O7:β AgVO3. After treatment, the culture medium was removed, 200
μL of neutral red solution (30 μg/mL) was added and the cells were incubated for 2
hours.
After this time, the cells were washed with PBS 1X and the neutral red with 200 μL of
ethanol solution 50% (v/v) and acetic acid 1% (v/v) was eluded followed by incubation
for 10 min at room temperature. Absorbance was measured at 540 nm. All tests were
conducted in triplicate. The calculation used to present the results was: