Non Animal Testing, Alternative Test Methods, In Vitro Toxicology, IIVS | NewsType Posters & Presentations
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Posters & Presentations

The 3D Human Reconstructed Skin Micronucleus Assay (RSMN) Using the EpiDerm Tissue: Validation and Application to the Safety Assessment of Cosmetics Ingredients

March 30, 2017

Regulatory restrictions on animal use have increased the reliance of risk assessors and regulators on in vitro test systems. Ideally, tissue-based assays could replace the animal studies as follow-up tools to verify results from standard in vitro assays. The RSMN assay combines the EpiDerm™ 3D reconstructed skin (RS) model with the micronucleus (MN) assay to provide a more realistic model for evaluating the genotoxic potential of dermally applied chemicals or products, such as cosmetics.

In Vitro Models Available for Testing of ENDS

February 20, 2017

Presented at the National Academies of Sciences, Engineering, and Medicine's (NAS) workshop on electronic nicotine delivery systems (ENDS), e-cigarettes.

Case Study: Applying GLPs to New, Non-Animal Methodologies

January 26, 2017

Presented at the 2nd Good Laboratory Practice Compliance Summit, January 26-27, 2017, Arlington, VA

Evaluation of the Validated In Vitro Skin Irritation Test (OECD TG 439) for the Assignment of EPA Hazard Categories

April 1, 2016

Progress within the in vitro toxicology field has made available testing platforms based on reconstructed tissue models that are validated to address the skin corrosion and irritation endpoints. While the validated in vitro assays can be used for the hazard identification of chemicals irritant to skin in accordance with the United Nations (UN) Globally Harmonized System (GHS) for Classification and Labeling of Chemicals, they were not calibrated to address the classification system used by the United States Environmental Protection Agency (US EPA).

Evaluation of New Solvents for the Use in the Multi‐Dose Reconstructed Human EpiDermis (RhE) Phototoxicity Assay

March 17, 2016

The phototoxic potential of test materials after exposure to UVA/visible light is evaluated by the 3T3 Neutral Red Uptake Phototoxicity Assay using Balb/c 3T3 mouse fibroblasts (OECD TG 432). To address challenges related to testing of finished products or materials that are not completely soluble, the Reconstructed Human EpiDermis (RhE) Phototoxicity Assay (INVITTOX Protocol 121) can be used as a stand‐alone or in a tiered approach. Water and sesame oil were recommended solvents for the RhE Assay, however additional solvents may be investigated on a case‐by‐case basis to accommodate a wider variety of test materials. Alternate solvents should first be assessed to ensure that they: 1) do not cause cytotoxicity; 2) do not diminish and/or inhibit phototoxic reactions; and 3) do not interfere with the UVA exposure. We investigated 5% DMSO in Hanks’ Balanced Salt Solution (HBSS), 5% acetone in HBSS, and polyethylene glycol (PEG) and their influence on the prediction of phototoxicity of chlorpromazine, a known phototoxicant. Duplicate tissues (EpiDerm™ from MatTek Corporation, Ashland, MA) were treated with each group for 24 hours, followed by a UVA (~6 J/cm2) or dark exposure, and then a 21 hour post‐exposure period before viability assessment using MTT dye. The assay positive control, 0.02% chlorpromazine in HBSS containing 1% DMSO, was tested concurrently. A test material was considered to have phototoxic potential if it induced a ≥30% difference in viability between tissues exposed to UVA as compared to the dark‐exposed tissues. Our data showed that the solvents performed in a comparable manner to the assay negative control (HBSS) and they did not induce significant toxicity when the viability of the UVA‐exposed and dark‐exposed tissues was compared: 92.9% and 104.8% (5% DMSO), 94.0% and 97.5% (5% acetone), and 81.3% and 90.0% (PEG), respectively. The difference between the viability of the UVA‐exposed or dark‐exposed tissues after treatment with 0.02% chlorpromazine dissolved in 1% DMSO, in 5% DMSO, and in PEG was 60.0%, 59.7%, and 72.5%, respectively. These data indicate that the new solvents we investigated were suitable for use in the detection of phototoxicity. The evaluation of 0.02% chlorpromazine in 5% acetone, as well as additional solvents (e.g. ethanol), is currently ongoing. Our future investigations will concentrate on the assessment of different solvents that can accommodate the phototoxicity testing of novel chemistries.