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Evaluating the toxicity of reactive dyes and dyed fabrics with the HaCaT cytotoxicity test

Klemola K., Pearson J., Lindström-Seppä P.

Autex Research Journal

2007

Vol. 7, no. 3

217--223

We investigated the cytotoxicity of reactive dyes and dyed fabrics using human keratinocyte HaCaT cells in vitro. The HaCaT cells were exposed to three monochlortriazinyl dyes: yellow, red and blue with different concentrations. The HaCaT cells were also exposed to water extracts of dyed fabrics. After 72 hours exposure, the protein contents of the samples compared to the protein contents of non-exposed cells were measured. The level of protein content indicates the viability of the cells. The mean inhibitory concentration values (IC50) showed the dye concentration when the protein content of the sample was 50% of the protein content of the non-exposed cells. The mean inhibitory concentration values (IC20) when the protein content of the samples was 80% were also measured. The IC20 values show the limiting value of toxicity. The IC50 values show whether samples are clearly toxic. The IC50 value for the yellow dye was 237µg/ml and the IC20 value was 78µg/ml. The IC50 for the red dye was 155µg/ml: the red dye caused adverse effects under the lowest dye concentration (28µg/ml). The IC50 value for the blue dye was 278µg/ml and the IC20 value was 112µg/ml. Cotton fabrics dyed using these same three reactive dyes were extracted with water and the extracts were analysed using the HaCaT cell line. The viability of the cells was good, the protein content of the samples being over 80% compared to the non-exposed cells. The HaCaT cell test indicated the toxicity of pure dyes; the dyed fabrics had no adverse effect. The human keratinocyte HaCaT cells seem to be a useful tool for the study of the purity/toxicity of dyes and other substances applied to textiles.

Evaluating the toxicity of reactive dyes and fabrics with the spermatozoa motility inhibition test

Klemola K., Honkalampi-Hämäläinen U., Liesivuori J., Pearson J., Lindström-Seppä P.

Autex Research Journal

2006

Vol. 6, no. 3

182--190

In this study, the toxicity of reactive dyes and dyed fabrics was investigated using spermatozoa cells in vitro. Boar semen was exposed to different concentrations of monochlorotriazinyl dyes: yellow, red and blue. The spermatozoa cells were also exposed to extracts of dyed fabrics. After 24 and 72 hours respectively, the viability of the cells was evaluated by microscopy. The mean inhibitor concentrations IC50, showing the concentration of the dye when half of the cells are dead compared to the control sample, were calculated from the viability values. After 24 hours' exposure, the IC50 value calculated for the yellow dye was 135µg/ml, and after 72 hours 60µg/ml. The IC50 value for the red dye was 124µg/ml after 24 hours, and 46µg/ml after 72 hours. The IC50 value for the blue dye after 24 hours was 127µg/ml. After 72 hours, the blue dye caused high toxicity: more than half the cells were dead. Cotton fabrics dyed using these three reactive dyestuffs were extracted by water and analysed by the spermatozoa motility inhibition test. The viability of the cells when exposed to fabric extracts was good. However, after 72 hours' exposure, the standard deviation and coefficient of variation values for cell viability of fabric extracts were large. The spermatozoa inhibition test indicated the toxicity of pure dyes, the dyed fabrics having no adverse effects. The spermatozoa test seems to be useful when screening different substances and when used in addition to other tests. The spermatozoa motility inhibition test can be used for textile material studies.