Synergy between ascorbate and α-tocopherol on fibroblasts in culture

Ascorbate and tocopherol are important antioxidants that protect cells against oxidative stress. The interaction of ascorbate and alpha -tocopherol in cells is difficult to detect as both ascorbate and alpha -tocopherol are unstable in vitro in a biological medium. We examined the interactions between human dermal fibroblasts, ascorbate and alpha -tocopherol to determine the effects of the vitamins on growth and cell viability. The interaction of ascorbate and alpha -tocopherol was studied in a fibroblast culture medium during 48h. Ascorbate and alpha -tocopherol were detected by fluorimetry after high-performance liquid chromatography (HPLC). Cell growth and cell viability were studied by cell numeration after trypan blue staining. The ascorbate concentration fell in presence of alpha -tocopherol in cell culture medium under all experimental conditions, with or without cells. Ascorbate partly protected alpha -tocopherol but only in presence of cells. Cell viability was preserved by alpha -tocopherol whereas ascorbate enhanced fibroblast growth. The synergy between ascorbate and alpha -tocopherol corresponds to a consumption of ascorbate which spares alpha -tocopherol but only in presence of cells. (C) 2001 Elsevier Science Inc. All rights reserved.