Mechanistic insight into ROS and neutral lipid alteration induced toxicity in the human model with fins (Danio rerio) by industrially synthesized titanium dioxide nanoparticles

Abstract

The toxicological impact of TiO₂ nanoparticles on the environment and human health has been extensively studied in the last few decades, but the mechanistic details were unknown. In this study, we evaluated the impact of industrially prepared TiO₂ nanoparticles on the biological system using zebrafish embryo as an <i>in vivo</i> model. The industrial synthesis of TiO₂ nanoparticles was mimicked on the lab scale using the high energy ball milling (HEBM) method by milling bulk TiO₂ particles for 5 h, 10 h, and 15 h in an ambient environment. The physiochemical properties were characterized by standard methods like field emission scanning electron microscopy (FESEM), dynamic light scattering (DLS), X-ray diffraction (XRD) and UV-Visible spectroscopy. <i>In vivo</i> cytotoxicity was assessed on zebrafish embryos by the evaluation of their mortality rate and hatching rate. Experimental and computational analysis of reactive oxygen species (ROS) induction, apoptosis, and neutral lipid alteration was done to study the effects on the cellular level of zebrafish larvae. The analysis depicted the change in size and surface charge of TiO₂ nanoparticles with respect to the increase in milling time. <i>In silico</i> investigations revealed the significant role of ROS quenching and altered neutral lipid accumulation functionalised by the molecular interaction of respective metabolic proteins in the cytotoxicity of TiO₂ nanoparticles with zebrafish embryos. The results reveal the hidden effect of industrially synthesized TiO₂ nanoparticle exposure on the alteration of lipid accumulation and ROS in developing zebrafish embryos. Moreover, the assessment provided a detailed mechanistic analysis of <i>in vivo</i> cytotoxicity at the molecular level.