Free radical formation in murine skin treated with tumour promoting organic peroxides

The generation of free radicals from tumour-promoting organic peroxides applied to intact murine skin samples has been studied by EPR spectroscopy using two techniques: first direct observation of ascorbyl radicals produced from reactions of peroxide-related radicals with ascorbate, an important endogenous antioxidant, and secondly, observation of radical adducts produced by spin-trapping. Free radical generation from tumour-promoting organic peroxides can be seen to occur in intact skin tissue through a one-electron reductive pathway, and takes place at sites including the viable cells of the epidermis and/or dermis. This radical generation is dependent upon the penetration of the skin by the peroxides, with the stratum corneum representing a major diffusional barrier to their penetration of skin. The technique of using ascorbyl radical measurement by EPR spectroscopy as a means of studying and quantifying radical production in intact tissues, developed in this work, may prove of much use in the study of many free radicals and their reactions in a wide range of biological systems, particularly skin. When combined with appropriate spin-trapping techniques, which enable identification of radical species and elucidation of their mechanisms of production, this enables the direct, real-time observation of radical reactions and mechanisms not previously possible in intact tissue samples.