Abstract
Nanoparticles (NPs) show various properties depending on their composition, size, and surface coating, which shape their interactions with biological systems. In particular, NPs have been shown to interact with immune cells, that represent a sensitive surveillance system of external and internal stimuli. In this light, in vitro models represent useful tools for investigating nano-bio-interactions in immune cells of different organisms, including invertebrates. In this work, the effects of selected types of NPs with different core composition, size and functionalization (custom-made PVP-AuNP and commercial nanopolystyrenes PS-NH2 and PS-COOH) were investigated in the hemocytes of the marine bivalve Mytilus galloprovincialis. The role of exposure medium was evaluated using either artificial seawater (ASW) or hemolymph serum (HS). Hemocyte morphology was investigated by scanning electron microscopy (SEM) and different functional parameters (lysosomal membrane stability, phagocytosis, and lysozyme release) were evaluated. The results show distinct morphological and functional changes induced in mussel hemocytes depending on the NP type and exposure medium. Mussel hemocytes may represent a powerful alternative in vitro model for a rapid pre-screening strategy for NPs, whose utilization will contribute to the understanding of the possible impact of environmental exposure to NPs in marine invertebrates.
Highlights
Due to the rapid expansion of production and use of nanoparticles (NPs) and their consequent release in different ecosystems, there is increasing concern on the utilization of alternative, affordable biological animal models for investigating nanosafety in environmental species
These studies underlined how mussel hemocytes represent a sensitive target for different NPs and their potential application as a starting point to more accurately design further studies that are relevant at the whole animal level
We investigated the in vitro responses of mussel hemocytes to different types of NPs in different exposure media with the aim of: (a) investigating the sensitivity of hemocytes to NPs with different surface characteristics and which characteristic of NPs are most potent to provoke the hemocyte response in physiological medium; defining optimal experimental conditions for a sensitive, alternative, and affordable biological in vitro model for a rapid prescreening strategy for NPs
Summary
Due to the rapid expansion of production and use of nanoparticles (NPs) and their consequent release in different ecosystems, there is increasing concern on the utilization of alternative, affordable biological animal models for investigating nanosafety in environmental species. Some types of NPs have been shown to associate with serum soluble components, organized into a “hard protein corona”, providing a specific biological identity for immune recognition and subsequent cellular responses [20,21] All these factors are important to consider in the evaluation of the possible biological impact of NPs on the cells of marine invertebrates, to properly utilize these cellular in vitro models as a suitable alternative strategy for testing the immunosafety of NPs in environmental organisms. Complementary experiments were performed to measure different functional immune markers, from functional integrity of lysosomes (lysosomal membrane stability) and phagocytic ability to extracellular defense mechanisms (e.g., lysozyme release and the production of ROS-reactive oxygen species)
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