Adverse Phototoxic Effect of Essential Plant Oils on NIH 3T3 Cell Line after UV Light Exposure.

AIM: Natural or artificial substances have become an inseparable part of our lives. It is questionable whether adequate testing has been performed in order to ensure these substances do not pose a serious health risk. The principal aim of our research was to clarify the potential risk of adding essential oils to food, beverages and cosmetic products. METHODS: The toxicity of substances frequently employed in cosmetics, aromatherapy and food industry (bergamot oil, Litsea cubeba oil, orange oil, citral) were investigated using cell line NIH3T3 (mouse fibroblasts) with/without UV irradiation. The MTT assay was used to estimate the cell viability. Reactive oxygen species (ROS) which are products of a number of natural cellular processes such as oxygen metabolism and inflammation were measured to determine the extent of cellular stress. DNA damage caused by strand breaks was examined by comet assay. RESULTS: MTT test determined EC50 values for all tested substances, varying from 0.0023% v/v for bergamot oil to 0.018% v/v for citral. ROS production measurement showed that UV radiation induces oxidative stress to the cell resulting in higher ROS production compared to the control and non-irradiated samples. Comet assay revealed that both groups (UV, without UV) exert irreversible DNA damage resulting in a cell death. CONCLUSIONS: Our findings suggest that even low concentrations (lower than 0.0464% v/v) of orange oil can be considered as phototoxic (PIF value 8.2) and probably phototoxic for bergamot oil (PIF value 4.6). We also found significant changes in the cell viability, the ROS production and the DNA after the cells were exposed to the tested chemicals. Even though these substances are widely used as antioxidants it should be noted that they present a risk factor and their use in cosmetic and food products should be minimized.

Acute and subacute oral toxicity of Litsea elliptica Blume essential oil in rats.

Litsea elliptica Blume has been traditionally used to treat headache, fever, and stomach ulcer, and has also been used as an insect repellent. The acute and subacute toxicities of L. elliptica essential oil were evaluated orally by gavage in female Sprague-Dawley rats. For the acute toxicity study, L. elliptica essential oil was administered in doses from 500 to 4000 mg/kg (single dose), and in the subacute toxicity test, the following doses were used: 125, 250, and 500 mg/kg, for 28 consecutive days. In the acute toxicity study, L. elliptica essential oil caused dose-dependent adverse behaviours and mortality. The median lethal dose value was 3488.86 mg/kg and the acute non-observed-adversed-effect level value was found to be 500 mg/kg. The subacute toxicity study of L. elliptica essential oil did not reveal alterations in body weight, and food and water consumptions. The haematological and biochemical analyses did not show significant differences between control and treated groups in most of the parameters examined, except for the hemoglobin, mean cell hemoglobin concentration, mean cell volume, mean cell hemoglobin, serum albumin, and serum sodium. However, these differences were still within the normal range. No abnormalities or histopathological changes were observed in the liver, pancreatic islet of Langerhans, and renal glomerulous and tubular cells of all treated groups. In conclusion, L. elliptica essential oil can be classified in the U group, which is defined as a group unlikely to present an acute hazard according to World Health Organization (WHO) classification.

Phototoxicity of essential oils intended for cosmetic use.

The aim of this study, linked-up with a previous study on bergamot oils, was the evaluation of phototoxic potential of essential oils (orange, lemon and Litsea cubeba), used as cosmetic ingredients. The applied tiered testing strategy included chemical analysis of the substances (by means of capillary gas chromatography/mass spectrometry), in vitro 3T3 NRU phototoxicity test and EpiDerm™ skin phototoxicity test. In order to clarify the situation in man, the highest non-phototoxic/non-cytotoxic concentrations and concentrations 10 x lower (safety factor 10) were tested xin vivo by means of human skin photopatch test in a limited group of human volunteers. The study revealed, that phototoxicity of the essential oils was dependent on the content of photoactive components and the solvent used. The highest non-phototoxic concentrations obtained by the skin model assay proved to be a useful starting point for subsequent confirmatory human photopatch test aimed to identify safe concentration for human use. However, the highest non-phototoxic concentration obtained in the skin model assay cannot be applied directly for human practice (3 of 8 tested oils evoked a phototoxic reaction). A safety factor of 10 should be applied for extrapolation of experimental data from the skin model assay to man.