Assessment of embryotoxicity of compounds in cosmetics by the embryonic stem cell test

The new EU legislations for cosmetics (Seventh Amendment) have laid down deadlines for the replacement of animal tests in cosmetics. This policy stimulates the acceptance of in vitro approaches to test embryotoxic potentials of compounds in cosmetics products. The embryonic stem cell test (EST) designed by The European Centre for the Validation of Alternative Methods (ECVAM) is currently the most promising in vitro assay to predict the embryotoxic potential of compounds. In this study, six selected compounds (hydroquinone, eugenol, dibutyl phthalate, antimony (III) oxide, neodymium (III) nitrate hydrate, melamine) formerly involved in cosmetic products were selected to test their embryotoxic potentials by the EST. 5-Fluorouracil and penicillin G were separately set as positive and negative control. The embryotoxic potential was determined by the prediction model (PM), which was calculated from three endpoints, the IC50 3T3, IC50 ES, and ID50. Hydroquinone, eugenol, and antimony (III) oxide indicated with strong embryotoxicity, while dibutyl phthalate, neodymium (III) nitrate hydrate, and melamine exhibited a weak embryotoxicity. These results may provide a valuable attempt to expand the application of EST to the cosmetics field.

[1]  J. Huff,et al.  Urolithiasis and bladder carcinogenicity of melamine in rodents. , 1984, Toxicology and applied pharmacology.

[2]  H. Spielmann,et al.  Reproduction and development. , 1998, Environmental health perspectives.

[3]  T. Mosmann Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[4]  A. Ozkul,et al.  Effect of hydroquinone (HQ) on the development of chick embryos. , 1994, Drug and chemical toxicology.

[5]  Elke Genschow,et al.  Embryotoxicity Screening Using Embryonic Stem Cells in vitro: Correlation to in vivo Teratogenicity , 1999, Cells Tissues Organs.

[6]  J. Rueff,et al.  Genotoxicity and endoreduplication inducing activity of the food flavouring eugenol. , 2006, Mutagenesis.

[7]  A. K. Peters,et al.  Automated analysis of contractility in the embryonic stem cell test, a novel approach to assess embryotoxicity. , 2008, Toxicology in vitro : an international journal published in association with BIBRA.

[8]  P. Steinberg,et al.  An in vitro embryotoxicity assay based on the disturbance of the differentiation of murine embryonic stem cells into endothelial cells. I: Establishment of the differentiation protocol. , 2007, Toxicology in vitro : an international journal published in association with BIBRA.

[9]  P. Foster,et al.  Pathogenesis of Male Reproductive Tract Lesions from Gestation Through Adulthood Following in Utero Exposure to Di(n-butyl) Phthalate , 2003, Toxicologic pathology.

[10]  R. Tyl,et al.  The induction of bladder stones by terephthalic acid, dimethyl terephthalate, and melamine (2,4,6-triamino-s-triazine) and its relevance to risk assessment. , 1985, Regulatory toxicology and pharmacology : RTP.

[11]  A M Wobus,et al.  Embryonic stem cells as an in vitro model for mutagenicity, cytotoxicity and embryotoxicity studies: present state and future prospects. , 2001, Toxicology in vitro : an international journal published in association with BIBRA.

[12]  Susanne Bremer,et al.  Validation of the Embryonic Stem Cell Test in the International ECVAM Validation Study on Three In Vitro Embryotoxicity Tests , 2004, Alternatives to laboratory animals : ATLA.

[13]  Robert E Chapin,et al.  Assessment of the Embryonic Stem Cell Test and application and use in the pharmaceutical industry. , 2008, Birth defects research. Part B, Developmental and reproductive toxicology.

[14]  K. Imai,et al.  In vitro embryotoxicity testing of metals for dental use by differentiation of embryonic stem cell test , 2006, Congenital anomalies.

[15]  Horst Spielmann,et al.  Screening Chemicals for Reproductive Toxicity: The Current Alternatives , 1995 .

[16]  A. Blacker,et al.  A study of developmental toxicity of hydroquinone in the rabbit. , 1992, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[17]  C. Laymon A. study , 2018, Predication and Ontology.

[18]  N. Z. Zur Nieden,et al.  Molecular multiple endpoint embryonic stem cell test--a possible approach to test for the teratogenic potential of compounds. , 2004, Toxicology and applied pharmacology.

[19]  Y. Ji,et al.  Toxicological studies on safety of rare earths used in agriculture. , 1988, Biomedical and environmental sciences : BES.

[20]  N Loprieno,et al.  Benzene and the genotoxicity of its metabolites. I. Transplacental activity in mouse fetuses and in their dams. , 1988, Mutation research.

[21]  Aldert H Piersma,et al.  Validation of the Rat Limb Bud Micromass Test in the International ECVAM Validation Study on Three In Vitro Embryotoxicity Tests , 2004, Alternatives to laboratory animals : ATLA.

[22]  A Léonard,et al.  Mutagenicity, carcinogenicity and teratogenicity of antimony compounds. , 1996, Mutation research.

[23]  A. Piersma,et al.  Development of prediction models for three in vitro embryotoxicity tests in an ECVAM validation study. , 2000, In vitro & molecular toxicology.

[24]  J. Furr,et al.  Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat. , 2000, Toxicological sciences : an official journal of the Society of Toxicology.

[25]  C. Price,et al.  Evaluation of the developmental toxicity of isoeugenol in Sprague-Dawley (CD) rats. , 2001, Toxicological sciences : an official journal of the Society of Toxicology.

[26]  R. Kruse,et al.  Decrease in Anogenital Distance among Male Infants with Prenatal Phthalate Exposure , 2005, Environmental health perspectives.

[27]  Aldert H Piersma,et al.  Validation of the Postimplantation Rat Whole-embryo Culture Test in the International ECVAM Validation Study on Three In Vitro Embryotoxicity Tests , 2004, Alternatives to laboratory animals : ATLA.

[28]  Margino Steemans,et al.  Evaluation of the embryotoxic potency of compounds in a newly revised high throughput embryonic stem cell test. , 2008, Toxicological sciences : an official journal of the Society of Toxicology.

[29]  Aldert Piersma,et al.  The ECVAM International Validation Study on In Vitro Embryotoxicity Tests: Results of the Definitive Phase and Evaluation of Prediction Models , 2002, Alternatives to laboratory animals : ATLA.

[30]  Jin Yj,et al.  Toxicological studies on safety of rare earths used in agriculture. , 1988 .

[31]  M K Smith,et al.  A selection of candidate compounds for in vitro teratogenesis test validation. , 1983, Teratogenesis, carcinogenesis, and mutagenesis.

[32]  H. Spielmann,et al.  The embryonic stem cell test (EST), an in vitro embryotoxicity test using two permanent mouse cell lines : 3T3 fibroblasts and embryonic stem cells , 1997 .

[33]  S. Bremer,et al.  Embryotoxicity hazard assessment of cadmium and arsenic compounds using embryonic stem cells. , 2008, Toxicology.