Abstract

In order to clarify that squalene monohydroperoxide (SQOOH) correlates with changes in morphology through cytotoxicity and establish in vitro evaluation of the cytotoxicity of lipid hydroperoxide, the effect of SQOOH on cytotoxicity and morphology in normal human epidermal keratinocytes (NHEK(B)) and the Gunze three-dimensional cultured human skin model (Vitrolife-skin) was investigated. Additionally, the effect of radical scavengers (mannitol, vitamin C and (+)-catechin) on the cytotoxicity in Vitrolife-skin was studied. The level of lipid hydroperoxide (phoshatidylcholine hydroperoxide: PCOOH) in the cellular membrane was increased with the concentration of SQOOH, and the rise in cytotoxicitv in NHEK(B) was associated with changes to the cellular membrane. A concentration-dependent and protective effect on the increase in cytotoxicity and PCOOH content was observed. To clarify the effect of SQOOH on the release of cytokine from cells, IL-2 level from NHEK(B) and Vitrolife-skin were investigated. IL-2 release from the cells was enhanced by SQOOH and increased at a non-cytotoxic dose. These results suggest that the increase in lipid hydroperoxides resulting from the auto-oxidation of lipids within cellular membranes in the presence of SQOOH correlates with changes in morphology due to cytotoxicity. SQOOH enhanced the release from cells at a non-cytotoxic dose. A method for assessing the protective effect on the cytotoxicity of lipid hydroperoxides using cells would be useful for in vitro evaluation of the cytotoxicity.

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