In vivo nanotoxicity testing using the zebrafish embryo assay

Abstract

Nanoparticles are increasingly used for biomedical purposes. Many different diagnostic and therapeutic applications are envisioned for nanoparticles, but there are often also serious concerns regarding their safety. Given the fact that numerous new nanomaterials are being developed every day, and that not much is known about the long-term toxicological impact of exposure to nanoparticles, there is an urgent need to establish efficient methods for nanotoxicity testing. The zebrafish (Danio rerio) embryo assay has recently emerged as an interesting 'intermediate' method for nanotoxicity screening, enabling (semi-) high-throughput analyses in a system significantly more complex than cultured cells, but at the same time also less 'invasive' and less expensive than large-scale biocompatibility studies in mice or rats. The zebrafish embryo assay is relatively well-established in the environmental sciences, but it has not yet gained wide notice in the nanomedicine field. Using prototypic polymeric drug carriers, gold-based nanodiagnostics and nanotherapeutics, and iron oxide-based nanodiagnostics, we here show that toxicity testing using zebrafish embryos is easy, efficient and informative, and faithfully reflects, yet significantly extends, cell-based toxicity testing. We therefore expect that the zebrafish embryo assay will become a popular future tool for nanotoxicity screening.

Curcumin-encapsulated nanoparticles as innovative antimicrobial and wound healing agent

“…25 This emerging assay has been used to evaluate toxicity of a broad array of nanoparticles, including other curcumin nanoformulations, 47 offering a more complex analysis compared to cell culture. 48 The highest concentration of 250 ppm evaluated in this study represents a worst case scenario, especially in terms of comparative concentrations in cosmetic products. The experimental duration is from 8 hours post-fertilization, when the embryo consists of only a few cells, to 120 hpf, which is the onset of free feeding.…”

Toxicological Risk Assessment of Emerging Nanomaterials: Cytotoxicity, Cellular Uptake, Effects on Biogenesis and Cell Organelle Activity, Acute Toxicity and Biodistribution of Oxide Nanoparticles

“…Iron overload, changes of gene expression and inhibition of acetylcholinesterase were proposed as causes for the observed neurotoxicity. In another study, SPIONs, which were non-toxic in vitro, were lethal in zebrafish embryos when used at concentrations higher than 10 mg/mL [80]. Regarding TiO 2 nanotubes, they are reported to have an excellent biocompatibility [84].…”

“…Recently, zebrafish models were used to evaluate the toxicity of various nanoparticles, including SPIONs and spherical TiO 2 [80][81][82][83]. Studies in zebrafish embryos point to toxicity using concentrations of iron oxide particles >10 mg/L resulting in increased mortality, hatching delay and malformations [81], showing the possibility of toxicity of SPIONs at elevated concentrations.…”

Decationized polyplexes for gene delivery

“…Furthermore, the neutral block copolymer pHP--PEG that forms decationized polyplexes showed excellent cytocompatibility upon incubation with HUVEC cells. Finally, an in vivo zebrafish nanotoxicity assay [21] also demonstrated that this polymer had a remarkable low teratogenicity and mortality profile, in great contrast with its biodegradable cationic counterpart ( Figure 2B) [22].…”