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In vitro acute toxicity assessment of tire tread particles using rainbow trout cell lines of gill and intestine
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Researchers tested the toxicity of tire and road wear particles (TRWP) on rainbow trout gill and intestine cells in a lab setting. TRWP extract was toxic to both cell types at higher concentrations, suggesting these particles — a major component of microplastic pollution in roads and stormwater — pose a threat to aquatic life. Tire wear particles enter waterways through rain runoff and are a significant but underregulated source of aquatic contamination.
Tire and road wear particles (TRWP) are generated by the abrasion of tires while driving, and recent questions were raised in regards to their potential contribution to microplastics released into the aquatic environment and their potential toxicological impacts. Our study aimed to determine the toxicity of TRWP and associated chemicals to fish using two rainbow trout (Oncorhynchus mykiss) cell lines representing the gill (RTgill-W1) and the intestinal (RTgut-GC) epithelium. Acute exposure to cryogenically milled tire tread (CMTT) particles and CMTT digestate following in vitro digestion was investigated. To obtain the effective concentrations (EC50) causing a 50% cell viability loss, tests were performed in accordance with OECD TG249: cell viability was assessed after 24 hours exposure using a multiple-endpoint assay indicative of cell metabolic activity, membrane integrity and lysosome integrity. Finally, chemical composition of the exposure medium was analyzed to assess which chemicals could be responsible for the observed acute effects. Our results show that metabolic activity was the most sensitive indicator of CMTT and CMTT digestate toxicity. EC50 values after direct exposure to RTgill-W1 and RTgutGC cells were estimated at 2.01 g/L and 3.80 g/L, respectively. 2-Mercaptobenzothiazole was found at the highest concentration among organic chemicals leaching out from the particles. It could be a major contributor to the loss in metabolic activity based on its toxicity to rainbow trout in vivo, although additional particle or mixture effects cannot be ruled out at this point. For digestate toxicity, EC50 was determined at 12.15 g of tire particle/L of digestive fluid. The observed toxicity could be due to high amounts of Zn, N-(1,3-dimethylbutyl)-N′-phenyl-p-phenylenediamine (6PPD), 1,3-Diphenylguanidine (DPG) or 2-Mercaptobenzothiazole leaching out during in vitro digestion. Traces of 6PPD-quinone were also detected in the exposure medium, but additional cytotoxicity assays using the RTgill-W1 cell line showed no significant toxicity of this compound up to 3 mg/L. Overall, our in vitro toxicity data agree with reports of acute fish (in vivo) toxicity and show that acutely toxic concentrations of CMTT are well above concentrations measured in river water (0.3 to 4 mg/L). However, we determined the highest no-observed-effect concentration at 10 mg/L, while up to 100 mg/L were measured in road runoffs.
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Evaluation of tire tread particle toxicity to fish using rainbow trout cell lines
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Researchers tested how tire and road wear particles — tiny fragments shed from vehicle tires — affect fish cells representing gill and intestinal tissue. They found toxicity was mainly driven by chemicals leaching out of the particles, including zinc and several rubber additives, with digestion conditions increasing how much of these chemicals were released.
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Researchers exposed rainbow trout sperm cells to tire rubber-based microplastic particles at various concentrations and measured the effects on sperm quality. They found that the microplastic leachates reduced sperm velocity and movement parameters in a dose-dependent manner and decreased antioxidant enzyme activity. The study suggests that tire-derived microplastics may pose a reproductive health risk to freshwater fish species.
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This systematic review examined tire wear particles — a type of microplastic created as tires wear down on roads — as an emerging water pollutant. These particles wash into rivers and oceans through stormwater runoff and contain toxic chemicals that harm aquatic organisms. Since tire wear is one of the largest sources of microplastic pollution, this is relevant to anyone living near roads or consuming seafood.
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Researchers studied how chemicals that leach from tire wear particles affect developing zebrafish, a common lab organism. The tire particle chemicals damaged eye development and impaired swimming behavior, even at concentrations found in the real environment. Since tire wear is one of the largest sources of microplastic pollution, these findings raise concerns about the ecological impact of road runoff on aquatic life.
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