A holistic study on potential toxic effects of carboxylated multi-walled carbon nanotubes (MWCNTs-COOH) on zebrafish (Danio rerio) embryos/larvae.

Multi-walled carbon nanotubes (MWCNTs) have widespread use in industrial and consumer products and great potential in biomedical applications. This leads to inevitably their release into the environment and the formation of their toxic effects on organisms. These effects can change depending on their physicochemical characteristics. Therefore, the toxicological findings of MWCNTs are inconsistent. Their toxicities related to surface modification have not been elucidated in a holistic manner. Hence, this study was conducted to clarify their potential toxic effects on zebrafish embryos/larvae in a comprehensive approach using morphologic, biochemical and molecular parameters. Zebrafish embryos were exposed to 5, 10, 20 mg/L doses of MWCNTs-COOH at 4 h after fertilization and grown until 96 hpf. Physiological findings demonstrated that they induced a concentration-dependent increase in the mortality rate, delayed hatching and decrease in the heartbeat rate. Moreover, it caused abnormalities including yolk sac edema, pericardial edema, head, tail malformations, and vertebral deformities. These effects may be due to the alterations in antioxidant and immune system related gene expressions after their entry into zebrafish embryo/larvae. The entry was confirmed from the evaluation of Raman spectra collected from the head, yolk sac, and tail of control and the nanotube treated groups. The gene expression analysis indicated the changes in the expression of oxidative stress (mtf-1, hsp70, and nfkb) and innate immune system (il-1ß, tlr-4, tlr-22, trf, and cebp) related genes, especially an increased in the expression of the hsp70 and il-1ß. These findings proved the developmental toxicities of MWCNTs-COOH on the zebrafish embryos/larvae.