ABSTRACT
To investigate the neurotoxic effects caused by the novel drug rutin trihydrate conjugated zinc oxide nanoparticles (RT-ZnONPs) in the zebrafish larvae model. By diluting the stock solution a test solution was prepared for the exposure group, containing 40 µg/ml concentrations of the novel drug rutin trihydrate conjugated zinc oxide nanoparticles. Maintenance and Collection of Zebrafish and Eggs: Adult zebrafish were procured from a nearby breeder and kept in acclimatization. Fishes were kept in a breeding tank with an equal ratio of males to females. The eggs were collected, washed in E3 medium, and placed into well plates for exposure to 40 μg/ml of rutin trihydrate-conjugated zinc oxide. Subsequently, they were subjected to 0.7mm hydrogen peroxide induction. Locomotory Behavior Analysis: Larvae's swimming behavior was observed at 6 dpf. Groups of 3 larvae from each group were kept in an ice cube tray containing 3ml of E3 medium for one minute. Larval movements were recorded using a commercial smartphone camera in a quiet environment at the beginning of the light cycle. The recorded video was then analyzed using UMA Tracker software. All experiments were conducted in triplicate. Statistical analysis was performed using the Statistical Packages for the Social Sciences (SPSS) software to identify the significance between the three groups. The parameters were set to a confidence level of 95%, a threshold of 0.05, a power of 80% using G power, and an enrollment ratio of 1. The locomotor behavior of larvae was found to be significantly increased in the rutin trihydrate conjugated zinc oxide nanoparticles treatment group (30.66±3.59) compared to the H2O2 treated group (78.11±4.64), with a significance of P=0.000 (P<0.05). The novel drug RT-ZnONPs were treated in the zebrafish embryos for 2 to 6 hours post fertilization (hpf) followed by the embryos being exposed to H2O2 to induce neurotoxicity. When compared to the untreated group, it was observed that the novel drug rutin trihydrate zinc oxide nanoparticles treated group larvae covered 71% area, while H2O2 group larvae covered only 37% area, with a significance of p=0.000 (P<0.05). Based on observation, the ROS-induced neurotoxicity has been reduced in the RT-ZnONPs treatment group compared to the H2O2 group.
