Selection of an optimal culture medium and the most responsive viability assay to assess AgNPs toxicity with primary cultures of Eisenia fetida coelomocytes

García-Velasco, Nerea; Irizar, Amaia; Urionabarrenetxea, Erik; Scott-Fordsmand, Janeck J.; Soto, Manu

doi:10.1016/j.ecoenv.2019.109545

Cited by 16 publications

(12 citation statements)

References 49 publications

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“…Previous in vitro studies have shown that NPs can be taken up by earthworm immune cells (coelomocytes) [ 45 , 80 , 81 ], and can alter the expression of earthworm immune markers [ 45 , 46 ], leading to cellular toxicity [ 82 , 83 ]. In this study, in vivo exposure to metal biocides (in both metal salt and NP form) caused changes to microbiome structure, with several bacterial symbionts being negatively affected by exposure to copper.…”

Section: Discussionmentioning

confidence: 99%

The Effects of In Vivo Exposure to Copper Oxide Nanoparticles on the Gut Microbiome, Host Immunity, and Susceptibility to a Bacterial Infection in Earthworms

et al. 2020

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Nanomaterials (NMs) can interact with the innate immunity of organisms. It remains, however, unclear whether these interactions can compromise the immune functioning of the host when faced with a disease threat. Co-exposure with pathogens is thus a powerful approach to assess the immuno-safety of NMs. In this paper, we studied the impacts of in vivo exposure to a biocidal NM on the gut microbiome, host immune responses, and susceptibility of the host to a bacterial challenge in an earthworm. Eisenia fetida were exposed to CuO-nanoparticles in soil for 28 days, after which the earthworms were challenged with the soil bacterium Bacillus subtilis. Immune responses were monitored by measuring mRNA levels of known earthworm immune genes. Effects of treatments on the gut microbiome were also assessed to link microbiome changes to immune responses. Treatments caused a shift in the earthworm gut microbiome. Despite these effects, no impacts of treatment on the expression of earthworm immune markers were recorded. The methodological approach applied in this paper provides a useful framework for improved assessment of immuno-safety of NMs. In addition, we highlight the need to investigate time as a factor in earthworm immune responses to NM exposure.

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Section: Discussionmentioning

confidence: 99%

The Effects of In Vivo Exposure to Copper Oxide Nanoparticles on the Gut Microbiome, Host Immunity, and Susceptibility to a Bacterial Infection in Earthworms

et al. 2020

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“…On the other hand, isolated coelomocytes of E. fetida cultivated in RPMI-1640 medium showed an LC 50 value of 6 µg/mL (Garcia-Velasco et al 2019). Coelomocytes cultivated in L-15 medium, on the other hand, showed a much higher resistance to Ag nanoparticles (LC 50 > 100 µg/mL) (Garcia-Velasco et al 2019). Differences between the studies may stem from differences in particle size as well as different cell types used and therefore non-comparable cellular reactions.…”

Section: Ecotoxicity Testing Using Primary Intestinal Cells Of E Fetidamentioning

confidence: 93%

Ex Vivo Cultivation Of Primary Intestinal Cells From Eisenia Fetida As Basis For Ecotoxicological Studies

Riedl

Völkl

Holzinger

et al. 2021

Preprint

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The earthworm Eisenia fetida is a commonly used model organism for unspecific soil feeders in ecotoxicological studies. Its intestinal cells are the first to encounter possible pollutants co-ingested by the earthworm, which makes them prime candidates for studies of toxic effects of environmental pollutants on the cellular as compared to the organismic level. Here, cells are isolated directly from the intestine, maintaining > 90% viability during subsequent short-time cultivations (up to 144 h). Exposure to established toxins comprising silver nanoparticles and metal ions (Cu2+, Cd2+) induced a significant decrease in the metabolic activity of the cells. In presence of microplastic particles (MP particles), namely 0.2, 0.5, 2.0, and 3.0 µm diameter polystyrene (PS) beads as well as 0.5 and 2.0 µm diameter polylactic acid (PLA) beads, no active uptake and no effect on the metabolic activity of the cells was observed. This suggests a tissue rather than cell related basis for the previously observed ecotoxicological effects of MP in case of Eisenia fetida.

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“…Immediately upon collection, the coelomocytes need to be stabilized in a culture medium in order preserve cell viability. Recent studies show that RPMI 1640 medium is the optimal medium for earthworm coelomocytes culturing, as well as the assessment of NP toxicity towards coelomocytes [135,136]. Critical for the successful culturing of coelomocytes is the adjustment of osmolality of the medium so that it reflects that of the coelomic fluid [137][138][139].…”

Section: Nonmammalian In Vitro Assaysmentioning

confidence: 99%

Cross-Species Comparisons of Nanoparticle Interactions with Innate Immune Systems: A Methodological Review

et al. 2021

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Many components of the innate immune system are evolutionarily conserved and shared across many living organisms, from plants and invertebrates to humans. Therefore, these shared features can allow the comparative study of potentially dangerous substances, such as engineered nanoparticles (NPs). However, differences of methodology and procedure between diverse species and models make comparison of innate immune responses to NPs between organisms difficult in many cases. To this aim, this review provides an overview of suitable methods and assays that can be used to measure NP immune interactions across species in a multidisciplinary approach. The first part of this review describes the main innate immune defense characteristics of the selected models that can be associated to NPs exposure. In the second part, the different modes of exposure to NPs across models (considering isolated cells or whole organisms) and the main endpoints measured are discussed. In this synergistic perspective, we provide an overview of the current state of important cross-disciplinary immunological models to study NP-immune interactions and identify future research needs. As such, this paper could be used as a methodological reference point for future nano-immunosafety studies.

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Selection of an optimal culture medium and the most responsive viability assay to assess AgNPs toxicity with primary cultures of Eisenia fetida coelomocytes

Cited by 16 publications

References 49 publications

The Effects of In Vivo Exposure to Copper Oxide Nanoparticles on the Gut Microbiome, Host Immunity, and Susceptibility to a Bacterial Infection in Earthworms

The Effects of In Vivo Exposure to Copper Oxide Nanoparticles on the Gut Microbiome, Host Immunity, and Susceptibility to a Bacterial Infection in Earthworms

Ex Vivo Cultivation Of Primary Intestinal Cells From Eisenia Fetida As Basis For Ecotoxicological Studies

Cross-Species Comparisons of Nanoparticle Interactions with Innate Immune Systems: A Methodological Review

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