Cell culture methods are being developed to assess the dermal toxicity (irritancy and corrosion) of chemicals. These in vitro methods are being validated to categorize chemicals as irritating or non-irritating to humans. Currently, these cell culture tests are useful to assist in the ranking of chemicals for irritancy, but they are not useful for quantitative risk assessment for two reasons. First, for volatile chemicals the amount of chemical in the media that the cells are exposed to may decrease with exposure time. Also, effective concentrations such as EC(50) and IC(50) are reported as the concentrations in the media not the skin tissue/cells. We have developed an in vitro approach for dermal toxicity testing of volatile chemicals that avoids these problems. Using sealed vials lacking a headspace, dermal equivalents (dermal fibroblasts in a collagen matrix) were exposed to culture medium containing a test chemical (m-xylene) and compared to a traditional open well culture system. We found that about 90% of the m-xylene was lost from the open well plates and the viability was 4-6 times greater than in the closed system. Partition coefficients were measured and used to estimate the m-xylene concentration in the fibroblasts. The EC(50) for m-xylene in the dermal equivalents was 833.13+/-35.33 microg m-xylene per gram of fibroblasts. This method will provide an effective approach to relate target cell chemical concentration to cellular responses. Based on this method, a biologically-based mathematical model could be used to determine an equivalent external dose for a specific toxic end point.
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