Manoj Kumar TM
Soumya Balakrishnana
Suresh Kunjiramana
Manomi Sarasan
Jason Tyler Magnuson
Jayesh Puthumana
Sreevidya CP
2025
<p><span>Bisphenol A (BPA) is a widely used industrial compound found in polycarbonate plastics, epoxy resin, and various polymer materials, leading to its ubiquitous presence in the environment. The toxicity of BPA to aquatic organisms has been well documented following </span><i>in vivo</i><span> exposure scenarios, with known cytotoxic and endocrine-disrupting effects. As such, BPA was used in this study as a well-characterized chemical to implement more ethical and resource-efficient scientific practices in toxicity testing through new approach methods (NAMs). Due to the frequent use of </span><i>Daphnia</i><span> spp. as a model organism in toxicology, we developed an </span><i>in vitro</i><span> cell culture system from </span><i>Daphnia magna</i><span> embryos, with optimized medium to support cell longevity. The cultures were maintained for up to two months, demonstrating their stability and suitability for cytotoxicity studies. Using this novel system, lethal concentration 50 (LC</span><sub>50</sub><span>) values were determined at the 24 and 48 h time points following BPA exposure. Subsequently, oxidative stress, endocrine disruption, and DNA damage were assessed through gene expression, activity assays, and a comet assay in BPA-exposed cells. LC</span><sub>50</sub><span> values of 52 µM and 20 µM BPA were calculated after 24 and 48 h exposures, respectively. BPA cells exposed to 20 and 52 µM had significantly increased GSH, GPx, and GST activity levels. mRNA expression analysis revealed significant upregulations in the expression of </span><i>hsp70, hsp90, gst, gpx, vtg1</i><span>, and </span><i>cyp4</i><span>, with downregulations of </span><i>sod, cat</i><span>, and </span><i>ecr</i><span> following BPA exposure. Furthermore, comet assays showed a significantly higher level of DNA damage induced by BPA compared to controls, with greater comet and tail lengths. This study established a novel </span><i>in vitro Daphnia</i><span> model, using BPA as a case study for determining toxic effects, further highlighting the importance and applicability of utilizing alternative methods in ecotoxicological research through reducing animal use.</span></p>
application/pdf
10.1016/j.aquatox.2024.107173
en
Elsevier
Establishment of a cell culture from Daphnia magna as an in vitro model for (eco)toxicology assays: Case study using Bisphenol A as a representative cytotoxic and endocrine disrupting chemical
article