The Center for Alternatives to Animal Testing is an academic center affiliated with the Division of Toxicological Sciences in the Department of Environmental Health Sciences of the Johns Hopkins University Bloomberg School of Public Health.

 

Johns Hopkins School of Public Health

Research Grants 1997-1998

Fluorescent Probing of Oxidative Stress and Antioxidant Efficacy in a Cell Culture Model

Valerian E. Kagan, PhD
University of Pittsburgh, Pittsburgh, Pennsylvania

Our broad long-term objective is to use a cell culture model that we have perfected for evaluating oxidative stress induced by commercial chemicals to now test the efficacy of antioxidants as protective (therapeutic) agents. Both cell systems and whole animal models for testing toxic agents thought to act through oxidative stress suffer from a major drawback: metabolic turnover and repair mechanisms for lipids (a major target of oxidative stress) are extremely efficient. Hence, it is difficult to determine whether oxidative stress has actually occurred.

A further drawback in both animal and cell systems is lack of sensitive measurement techniques. The combination of these two deficiencies means that, at present, extremely large quantities of cells or many animals must be used or, alternatively, extremely large, unphysiological doses of oxidant must be applied. Our perfected model eliminates both these drawbacks, thereby providing a model for testing putative oxidants and antioxidants that is superior both to current animal testing and to current cell culture techniques.

In our first year, we completed the development of the model for testing oxidants. In the second year, we wish now to use it for testing the antioxidants. Our approach and model to test the effectiveness of antioxidants will have the following unique features: