Table 1. Percentage changes (mean ± SD) in cell viability after 24 h of treatment with positive control.
Results – Cytotoxicity evaluation (MTT test) – EPSAN® C
Evaluation of cell viability
The results obtained are reported in charts and tables containing the measurements of cell viability (MTT assay) performed on HaCaT cells treated with the product to be tested and positive control (SLS), with respect to the negative control (untreated cells). Data are presented as mean ± standard deviation (SD).
In Figures 1. and in Table 1. graph and corrisponding data of cell viability of the positive control are shown. Data were processed using Photox v. 2.0 for IC50 calculation.
Figure 1. Cell viability after 24 h treatment of HaCaT cells with positive control (n=1; replicates=3)
| SLS (positive control) | |||||||||
| Sample (%) | 0 | 0.0039 | 0.0078 | 0.0156 | 0.0313 | 0.0625 | 0.125 | 0.25 | 0.5 |
| Cell viability (%) | 100 | 104.37 | 56.83 | 6.46 | 6.50 | 6.33 | 7.03 | 6.49 | 6.58 |
| Standard deviation | 18.5 | 7.64 | 2.80 | 0.17 | 0.14 | 0.14 | 0.31 | 0.20 | 0.36 |
| IC50 value | 1.20 | ||||||||
In Figures 2. and in Table 2. graph and corrisponding data of cell viability of the the tested product, EPSAN® C, are shown.
Figure 2. Cell viability after 24 h treatment of HaCaT cells with the tested product (n=2; replicates=3)
| EPSAN® C | |||||||||
| Sample (%) | 0 | 0.023 | 0.047 | 0.094 | 0.188 | 0.375 | 0.75 | 1.5 | 3 |
| Cell viability (%) | 100 | 91.82 | 82.20 | 83.94 | 78.72 | 66.23 | 59.37 | 37.22 | 10.22 |
| Standard deviation | 0 | 3.77 | 0.95 | 1.74 | 1.61 | 3.03 | 2.58 | 1.05 | 1.53 |
| IC50 value | 0.963 | ||||||||
Table 2. Percentage changes (mean ± SD) in cell viability after 24 h of treatment with the tested product.
The product shows a cytotoxic activity after 24h with 0.375% up to 3%. 0.188% and 0.094% are the first two non-cytotoxic concentrations (highlighted in red) which do not cause a cell mortality > 30%.
About IC50, the 0.008 calculated for SLS and 0.963 calculated for the product correspond to the scale expressed in percentage. In mg/ml these values are equal to 0.08 for SLS treatment and 9.63 for EPSAN® C treatment.
Correlating IC50 value with the cytotoxic/irritant effect according to the Table 3., SLS results to have a strong cytotoxic/irritant effect as a good positive control, while the product results to have no cytotoxic/irritant effect.
| IC50 value (mg/ml) | Effect |
| IC50 < 0.5 |
Strong cytotoxic/irritant effect
|
| 0.5 < IC50 < 1.5 |
Slight cytotoxic/irritant effect
|
| IC50 > 1.5 |
No cytotoxic/irritant effect
|
Table 3. Relation between IC50 value (mg/ml) and the possible cytotoxic/irritant effect.
