The biodistribution and immuno-responses of differently shaped non-modified gold particles in zebrafish embryos.

Important questions raised in (nano)ecotoxicology are whether biodistribution of nanoparticles (NPs) is affected by particle shape and to what extent local adverse responses are subsequently initiated. For nanomedicine, these same questions become important when the labeled NPs lose the labeling. In this study, we investigated the biodistribution patterns of gold nanoparticles (AuNPs) as well as immune-related local and systemic sublethal markers of exposure and behavioral assessment. Hatched zebrafish embryos were exposed to four differently shaped non-coated AuNPs with comparable sizes: nanospheres, nanorods, nano-urchins, and nano-bipyramids. Shape-dependent trafficking of the particles resulted in a different distribution of the particles over the target organs. The differences across the distribution patterns indicate that the particles behave slightly different, although they eventually reach the same target organs - yet in different ratios. Mainly local induction of the immune system was observed, whereas systemic immune responses were not clearly visible. Macrophages were found to take AuNPs from the body fluid, be transferred into the veins and transported to digestive organs for clearance. No significant behavioral toxicological responses in zebrafish embryos were observed after exposure. The trafficking of the particles in the macrophages indicates that the particles are removed via the mononuclear phagocytic system. The different ratios in which the particles are distributed over the target organs indicate that the shape influences their behavior and eventually possibly the toxicity of the particles. The observed shape-dependent biodistribution patterns might be beneficial for shape-specific targeting in nanomedicine and stress the importance of incorporating shape-features in nanosafety assessment.

Exploring uptake and biodistribution of polystyrene (nano)particles in zebrafish embryos at different developmental stages.

In ecotoxicology, it is continuously questioned whether (nano)particle exposure results in particle uptake and subsequent biodistribution or if particles adsorb to the epithelial layer only. To contribute to answering this question, we investigated different uptake routes in zebrafish embryos and how they affect particle uptake into organs and within whole organisms. This is addressed by exposing three different life stages of the zebrafish embryo in order to cover the following exposure routes: via chorion and dermal exposure; dermal exposure; oral and dermal exposure. How different nanoparticle sizes affect uptake routes was assessed by using polystyrene particles of 25, 50, 250 and 700nm. In our experimental study, we showed that particle uptake in biota is restricted to oral exposure, whereas the dermal route resulted in adsorption to the epidermis and gills only. Ingestion followed by biodistribution was observed for the tested particles of 25 and 50nm. The particles spread through the body and eventually accumulated in specific organs and tissues such as the eyes. Particles larger than 50nm were predominantly adsorbed onto the intestinal tract and outer epidermis of zebrafish embryos. Embryos exposed to particles via both epidermis and intestine showed highest uptake and eventually accumulated particles in the eye, whereas uptake of particles via the chorion and epidermis resulted in marginal uptake. Organ uptake and internal distribution should be monitored more closely to provide more in depth information of the toxicity of particles.

Comparability of behavioural assays using zebrafish larvae to assess neurotoxicity.

Testing of compounds for neurotoxicity has become increasingly important in recent years. It has been shown that neurological disorders like autism may be related to chemical exposures, which may play a crucial role in the progression of these diseases. Special attention has been be given to the substances causing developmental neurotoxicity as the developing nervous system is more vulnerable to impacts by chemicals than the adult nervous system. The zebrafish (Danio rerio) is a well-established model species in developmental biology and an emerging model in behavioural and neurological studies. Zebrafish larvae display numerous behavioural patterns highly similar to rodents and humans. Their physical characteristics make them well suited for automated high-throughput screening. In the last years, the number of behavioural studies conducted with zebrafish larvae has increased notably. The goal of this review is to provide an overview of behavioural assays commonly used to test substances for developmental neurotoxicity. Literature from 1995 to 2014 was reviewed and focussed on assays performed with zebrafish larvae younger than 7 days post fertilization (dpf). The behavioural tests were scrutinized, and parameters describing the different experimental setups were defined. In the next step, we investigated if differences in the experimental parameters alter the outcome of the test. In order to test the comparability of behavioural assays, we analysed several studies using ethanol, valproate and pentylenetetrazole as model substances. Based on our findings, we provide recommendations which could help improve future behavioural studies performed with zebrafish larvae.