Neurotoxicity Evaluation of Rutin Trihydrate vs. Metformin in Zebrafish Larvae: A Comparative Risk Assessment Study

Abstract

To examine the neurotoxicity induced by the novel drug Rutin trihydrate and metformin in the zebrafish larvae model. Test solutions for the exposure groups were prepared by diluting the stock solution with egg water. These solutions contained 30 µM metformin in combination with 9.5 µM of the innovative drug rutin trihydrate. Maintenance of zebrafish and egg collection: Breeding groups were chambered in a specific spawning tank in a male-to-female ratio of 1:1. The spawning tank is provided with a box and with mesh as the bottom base to collect the zebrafish embryos and to protect the zebrafish embryos from the adult fish. Embryotoxicity assay: This work includes 2 groups and a total sample size of 30. Group 1 underwent examination using the novel drug Rutin trihydrate, which exhibits solubility in 0.01% of Dimethyl sulfoxide (DMSO), while Group 2 was subjected to examination using metformin, soluble in 100µl of DMSO (Dimethyl sulfoxide) combined with 99% water, both assessments conducted on larval zebrafish. Embryos were segregated for each exposure in groups in petri plates with n= 15 embryos per plate and were exposed to the novel drug Rutin trihydrate from 4 - 96 hpf (Hours Post Fertilization). Other conditions required for the validation of this study were maintained as stated by the OECD (Organization for Economic All the experiments were carried out in triplicate. Statistical analysis: SPSS software facilitated the determination of statistical significance between the two groups. The parameters adhered to a confidential ratio of 95%, a threshold of 0.05, G power at 80%, and an enrolment ratio of 1. Results indicated a significant neuroprotective effect of rutin trihydrate-treated larvae (27.77±1.15) compared to metformin-treated larvae (18.0±0.68), displaying a statistical significance of p = 0.000 (p < 0.05).