Iron-induced Lipid Peroxidation in Human Skin-derived Cell Lines: Protection by a Red Orange Extract
F Morini1, F Dusatti1, F P Bonina2
1Department of Experimental Medicine, General Pathology Division, University of Genoa, Via L.B. Alberti 2, 16132 Genoa, Italy.
Alternatives to Laboratory Animals : ATLA
|November 25, 2014
Summary
This study shows that cultured human skin cells can be used to test chemical stress, like iron-induced lipid peroxidation. A red orange extract demonstrated significant antioxidant effects, supporting its use in cosmetic and drug development.
Area of Science:
- Biochemistry
- Cell Biology
- Dermatology
Background:
- Photodamage and photoprotection are well-studied in cell cultures.
- Limited data exist on in vitro skin cell responses to chemical stress, specifically iron-induced lipid peroxidation.
- Understanding these responses is crucial for developing new skincare and therapeutic agents.
Purpose of the Study:
- To investigate the susceptibility of human keratinocytes (NCTC 2544) and fibroblasts (HFFF2) to iron-induced lipid peroxidation.
- To evaluate the protective effects of a classical antioxidant mixture (Trolox C™ and Vitamin C) against oxidative stress.
- To assess the efficacy of a standardized red orange extract (ROE) as a potential antioxidant for skin cells.
Main Methods:
- Induction of lipid peroxidation using iron salts (FeSO4, Fe2(SO4)3) with histidine, ascorbate, or ADP.
- Quantification of lipid peroxidation by measuring malondialdehyde (MDA) content.
- Assessment of cell viability using neutral red uptake and tetrazolium salt reduction assays.
- Exposure of cell lines to varying concentrations of ROE and antioxidant mixtures.
Main Results:
- NCTC 2544 keratinocytes were more susceptible to lipid peroxidation than HFFF2 fibroblasts under specific iron-induced conditions.
- The iron/histidine model was selected for further antioxidant testing.
- Simultaneous addition of antioxidants and pro-oxidants maximized protection.
- ROE demonstrated significant, dose-dependent inhibition of MDA production, with over 90% cell viability maintained.
- ROE's active components include anthocyanins, hydroxycinnamic acid, and flavanone glycosides.
Conclusions:
- Cultured human skin cells exhibit differential susceptibility to chemically induced oxidative stress.
- The iron/histidine model provides a reliable method for assessing antioxidant efficacy in vitro.
- A standardized red orange extract possesses significant antioxidant properties beneficial for skin cells.
- These findings support the use of human skin cell lines for evaluating antioxidant ingredients in cosmetic and pharmaceutical development.
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