Acute, reproductive, and developmental toxicity of essential oils assessed with alternative in vitro and in vivo systems
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REPRODUCTIVE TOXICOLOGY
Acute, reproductive, and developmental toxicity of essential oils assessed with alternative in vitro and in vivo systems Peter Lanzerstorfer1,2 · Georg Sandner1 · Johannes Pitsch1,2 · Bianca Mascher2 · Tobias Aumiller3 · Julian Weghuber1,2 Received: 17 July 2020 / Accepted: 28 October 2020 © The Author(s) 2020
Abstract Essential oils (EOs) have attracted increased interest for different applications such as food preservatives, feed additives and ingredients in cosmetics. Due to their reported variable composition of components, they might be acutely toxic to humans and animals in small amounts. Despite the necessity, rigorous toxicity testing in terms of safety evaluation has not been reported so far, especially using alternatives to animal models. Here, we provide a strategy by use of alternative in vitro (cell cultures) and in vivo (Caenorhabditis elegans, hen’s egg test) approaches for detailed investigation of the impact of commonly used rosemary, citrus and eucalyptus essential oil on acute, developmental and reproductive toxicity as well as on mucous membrane irritation. In general, all EOs under study exhibited a comparable impact on measured parameters, with a slightly increased toxic potential of rosemary oil. In vitro cell culture results indicated a concentration-dependent decrease of cell viability for all EOs, with mean IC50 values ranging from 0.08 to 0.17% [v/v]. Similar results were obtained for the C. elegans model when using a sensitized bus-5 mutant strain, with a mean L C50 value of 0.42% [v/v]. In wild-type nematodes, approximately tenfold higher L C50 values were detected. C. elegans development and reproduction was already significantly inhibited at concentrations of 0.5% (wild-type) and 0.1% (bus-5) [v/v] of EO, respectively. Gene expression analysis revealed a significant upregulation of xenobiotic and oxidative stress genes such as cyp-14a3, gst-4, gpx-6 and sod-3. Furthermore, all three EOs under study showed an increased short-time mucous membrane irritation potential, already at 0.5% [v/v] of EO. Finally, GC–MS analysis was performed to quantitate the relative concentration of the most prominent EO compounds. In conclusion, our results demonstrate that EOs can exhibit severe toxic properties, already at low concentrations. Therefore, a detailed toxicological assessment is highly recommended for each EO and single intended application. Keywords Toxicological assessment · In vivo · C. elegans · HET-CAM · Essential oil
Introduction Peter Lanzerstorfer and Georg Sandner contributed equally to the manuscript. * Peter Lanzerstorfer peter.lanzerstorfer@fh‑wels.at * Julian Weghuber julian.weghuber@fh‑wels.at 1
Center of Excellence Food Technology and Nutrition, University of Applied Sciences Upper Austria, Stelzhamerstraße 23, 4600 Wels, Austria
2
Austrian Competence Centre for Feed and Food Quality, Safety and Innovation, Head Office: FFoQSI GmbH, Technopark 1C, 3430 Tulln, Austria
3
Delacon Biotechnik GmbH, Langwiesen 24, 4209 Engerwitzdorf, Austr
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