Investigating the dual functions of butylated hydroxytoluene, vitamin E and vitamin C as antioxidants and anti‐glycation agents in vitro: Implications for skin health

OBJECTIVES: Antioxidants are vital in skincare for neutralizing reactive oxygen species (ROS), which impact key cellular structures, such as cell nuclei and elastic fibres and can contribute to skin ageing. Oxidative stress, compounded by processes such as glycation, therefore leads to impaired cellular renewal and reduced skin elasticity. The effectiveness of antioxidants depends on their concentrations, making it essential to investigate their dosage-related benefits to optimize skincare formulations. This raises an important question regarding the reciprocal effects of antioxidants on glycation and whether their protective effects against macromolecular damage are dose-dependent. METHODS: This study evaluated the antioxidant and anti-glycation effects of three concentrations of butylated hydroxytoluene (BHT), vitamin E and vitamin C. Using the established quantitative assays and immunofluorescence, total antioxidant capacities, the intracellular ROS production, glycation levels and expression of cellular biomarkers were measured in dermal fibroblasts and three-dimensional skin models cultured with methylglyoxal (MGO). RESULTS: All three antioxidants showed a significant ability to inhibit the formation of intracellular ROS and glycation products induced by MGO. Notably, there were differences in the concentrations required to defend against glycation-induced damage. Whilst the linear dose responses were observed for ROS and glycation inhibition, the protective effect against cellular damage was moderate. The inverse dose-response relationships, particularly in terms of collagen fibre preservation, suggested that higher total antioxidant capacity could have enhanced protective effects. Vitamin C, in particular, exhibited the most pronounced benefits at lower concentrations, suggesting its potential as a key player in combating glycation damage. CONCLUSION: The potentially novel aspect of this research lies in the synergistic relationship between the modulation of oxidative stress and glycation. This relationship significantly depends on the concentration of the molecules involved and their antioxidant properties. These findings may lead to more refined approaches in formulating active ingredients tailored to individual needs in personalized skincare.