Abstract
Extensive exploitation of titanium dioxide nanoparticles (TiO2NPs) augments rapid release into the marine environment. When in contact with the body fluids of marine invertebrates, TiO2NPs undergo a transformation and adhere various organic molecules that shape a complex protein corona prior to contacting cells and tissues. To elucidate the potential extracellular signals that may be involved in the particle recognition by immune cells of the sea urchin Paracentrotus lividus, we investigated the behavior of TiO2NPs in contact with extracellular proteins in vitro. Our findings indicate that TiO2NPs are able to interact with sea urchin proteins in both cell-free and cell-conditioned media. The two-dimensional proteome analysis of the protein corona bound to TiO2NP revealed that negatively charged proteins bound preferentially to the particles. The main constituents shaping the sea urchin cell-conditioned TiO2NP protein corona were proteins involved in cellular adhesion (Pl-toposome, Pl-galectin-8, Pl-nectin) and cytoskeletal organization (actin and tubulin). Immune cells (phagocytes) aggregated TiO2NPs on the outer cell surface and within well-organized vesicles without eliciting harmful effects on the biological activities of the cells. Cells showed an active metabolism, no oxidative stress or caspase activation. These results provide a new level of understanding of the extracellular proteins involved in the immune-TiO2NP recognition and interaction in vitro, confirming that primary immune cell cultures from P. lividus can be an optional model for swift and efficient immune-toxicological investigations.
Highlights
Titanium dioxide nanoparticles (TiO2NPs) are very appealing nanomaterials for a variety of emerging applications
Because we found that cell-free medium contains an average protein concentration of about 7.093 ± 0.565 μg mL−1, we postulated that incubation of TiO2NPs within this medium would result in the adsorption of proteins onto the particle surface
Our results indicated a general healthy state of sea urchin immune cells in culture with TiO2NP exposure, which was not perceived as a stress inducer that was confirmed because it did not stimulate the release of the Hsp70 into the extracellular environment. p38 MAPK is a member of intracellular kinases that serves as focal point for diverse extracellular stimuli [45], which was used here as negative control
Summary
Titanium dioxide nanoparticles (TiO2NPs) are very appealing nanomaterials for a variety of emerging applications. TiO2NPs and Paracentrotus lividus Immunity use of TiO2NPs stimulated an outbreak of competing research (from molecules to ecosystems) aimed at delineating the safe limits of TiO2NPs use in occupational or environmental exposures This emphasizes the great level of interest concerning the biosafety of TiO2NPs. a complete and comprehensive understanding of the biosafety of TiO2NPs has not yet been achieved, due to large gaps of information in risk management, disposal, knowledge on the biomedical potentiality, interaction with many different biological factors (molecules) in environments and microenvironments, and due to ethical issues. New and more feasible approaches have resulted in the development of the novel in vitro–ex vivo methods to limit the use of living organisms [4] Based on this aim, we use primary cell cultures that accurately represent the biological tissue microenvironment in which cells reside, because cell–cell signaling remains preserved [5]. Primary cultures have become a much more appropriate tool for biotechnological applications and nano-safety/nano-toxicity investigations
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