Quantitation of cutaneous toxicity: an in vitro approach using skin organ culture

A short-term (48 hr) organ culture system for maintaining metabolically viable and structurally intact mouse skin has been developed. With the culture system, the responses of skin to in vitro physical and chemical insults were assessed by histologic examination and selected biochemical measurements in an attempt to establish a quantitative basis for the evaluation of cutaneous toxicity in vitro. In studies with mouse skin in organ culture following either freeze-thaw treatment (physical insult) or a single in vitro topical application of tributyltin chloride (10 to 1000 nmole/cm2), deleterious morphologic changes were observed. The degree of cellular injury was reflected by inhibition of in vitro incorporation of [3H]thymidine and [14C]leucine into epidermal DNA and protein, respectively; there also was leakage of intracellular enzymes into the culture medium in a dose and time related manner. These biochemical parameters are sensitive indicators of cellular injury; as such they may offer a simple, sensitive, and quantitative measure for assessing cutaneous toxicity of chemicals in vitro.