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Herbul black henna (hair dye) causes cardiovascular defects in zebrafish (Danio rerio) embryo model

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Herbul black henna (hair dye) have been widely used as cosmetic agents to temporarily change the color of hair. Their use was tremendously increased in the past decade. Especially, hair dye containing paraphenylenediamine (PPD) is extensively used globally because of its abundance and low cost. PPD, one of the main chemicals in hair dye, is known as a toxin. Hair dye has various adverse effects, including negative impacts on human health, especially during pregnancy and on aquatic environment. Although a large amount of dyes has been released into the environment, studies on environmental toxicity of these substances are still poorly understood. Hence, the main objective of this study is to evaluate the potential cardiovascular toxicological effects of hair dye on zebrafish (Danio rerio) embryo as a model. After exposing zebrafish embryos to various concentrations (100, 200, 300, 400, 500, and 600 μM), their cardiotoxicity and other teratogenic effects were examined for various exposure periods ranged from 24 to 96 hpf. As a result, the tested embryos could not survive over intervals of 48 h after 72 and 96 hpf at higher concentrations (300, 400, 500, and 600 μM), except the lower concentration cases (100 and 200 μM) some embryos were survived, respectively. However, hair dye increases mortality and decreases yolk stalk length, heart rate and severely affects heart looping in zebrafish embryos. In addition, the dye was observed to induce cardiovascular defects in transgenic Tg (fli1a:EGFP) and Tg (flk1:EGFP) zebrafish embryos at environmentally realistic dye concentrations. However, further study using bioassays is fundamentally required to predict the toxicity of hair dyes, aside from providing the information on their safe levels for living organisms. Therefore, the present results would be vital for the ecological risk assessment of hair dye, particularly for freshwater aquatic ecosystem and human health.

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The first author is grateful for the Korea Research Fellowship Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (KRF Grant No. 2016H1D3A1938252).


This work was supported by the National Natural Science Foundation of China (Grant No: 31400979) and Shandong Provincial Major Projects of Independent Innovation (2014ZZCX02105).

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Correspondence to Kechun Liu or Sang Joon Lee.

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Manjunatha, B., Han, L., Kundapur, R.R. et al. Herbul black henna (hair dye) causes cardiovascular defects in zebrafish (Danio rerio) embryo model. Environ Sci Pollut Res (2020). https://doi.org/10.1007/s11356-020-07762-z

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  • Hair dye
  • Teratogenic toxicity
  • Cardiotoxicity
  • Vascular defects
  • DNA damage
  • Zebrafish embryo