Evaluating the Cytotoxic Effect of the Arrow Root Extracted Sliver Nano Particles Using Zebra Fish Embryonic Sensitivity Test and Brine Shrimp Lethality Assay - An in vivo Study

Abstract

Introduction: Silver nanoparticles (AgNPs) have gained significant attention due to their promising applications in medicine, electronics, and environmental sciences. However, their cytotoxicity remains a concern, particularly when synthesized using plant extracts. This study evaluates the cytotoxic effects of silver nanoparticles synthesized using arrowroot extract (Maranta arundinacea) on two in vitro models: the zebrafish embryonic sensitivity test and the brine shrimp lethality assay.

Materials and Methods: Silver nanoparticles were synthesized by reducing silver nitrate (AgNO₃) with arrowroot extract, and their characteristics were determined through UV-Vis spectrophotometry and transmission electron microscopy (TEM). The cytotoxicity of the synthesized nanoparticles was assessed using two in vitro models: the zebrafish (Danio rerio) embryonic sensitivity test and the brine shrimp (Artemia salina) lethality assay. In the zebrafish assay, embryos were exposed to various concentrations of AgNPs, and developmental abnormalities, hatching rate, and survival were monitored. In the brine shrimp assay, larvae were exposed to different concentrations of AgNPs, and mortality rates were recorded to calculate the lethal concentration (LC50).

Results:  Both assays revealed concentration-dependent toxicity of the silver nanoparticles. In the zebrafish model, exposure to AgNPs resulted in a significant reduction in survival, delayed hatching, and morphological deformities such as abnormal body curvature and heart malformation. In the brine shrimp assay, a marked increase in mortality was observed, with the LC50 values calculated for different nanoparticle concentrations. The toxicity observed in both assays suggests that the arrowroot-extracted silver nanoparticles exhibit potent cytotoxic effects at higher concentrations.

Discussion: The results demonstrate that arrowroot-extracted silver nanoparticles possess significant cytotoxic potential, as evidenced by the developmental and survival impairments in zebrafish embryos and the high lethality in brine shrimp larvae. The concentration-dependent toxicity indicates that these nanoparticles may pose a risk to aquatic organisms and the environment. The mechanisms underlying this toxicity are likely related to oxidative stress and nanoparticle uptake, though further research is needed to elucidate the exact pathways. These findings contribute to the growing body of knowledge regarding the safety of silver nanoparticles synthesized from plant extracts.

Conclusion: This study highlights the cytotoxicity of silver nanoparticles synthesized using arrowroot extract and underscores the need for careful evaluation of their environmental and biological safety before widespread application. The zebrafish and brine shrimp assays provide valuable insights into the potential risks posed by silver nanoparticles, particularly in aquatic environments. Future research should focus on understanding the underlying mechanisms of toxicity and developing strategies to mitigate adverse effects, thereby ensuring the safe use of these nanoparticles in various fields.