Lack of in vitro protection by a common sunscreen ingredient on UVA-induced cytotoxicity in keratinocytes

As an extension of our previous investigations on sunscreen ingredients, the present work was aimed at assessing the possible protective effects of a common UVA-absorbing agent, Parsol 1789 (4-tert-butyl-4'-methoxydibenzoylmethane) in contact with human keratinocytes under UVA illumination. Cell viability was evaluated by determining lactate dehydrogenase (LDH) release, uptake of propidium iodide and fluorescein diacetate, total protein content and percentage of cell detachment. Apoptosis was detected by recognition of translocated phosphatidylserine using annexin V-FITC uptake. Oxidative stress was evaluated through the carboxy-H,DCFDA assay while the total oxyradical scavenging capacity (TOSC) assay was used for determining the total antioxidant capacity level in these cells. Lipid peroxidation was also assessed by checking hydroperoxide (HP) levels. The results obtained show that UVA exposure induces significant cell mortality, decrease in protein concentration, release of LDH, increase in apoptosis, oxidative stress and lipid peroxidation with a concomitant reduction in the response of the antioxidant cellular defense system. The presence of 10 muM Parsol 1789 did not minimize these UVA-induced effects, on the contrary, for some parameters measured such as lipid hydroperoxides, there was a significant enhancement. Furthermore, the presence of glutathione (GSH) alone decreased the level of ROS and lipid hydroperoxides, but in combination with Parsol 1789, this protective effect was reduced. The overall results indicate that the compound does not protect these cells from UVA exposure under our experimental conditions confirming previous findings on the lack of photoprotective efficiency of this sunscreen in contact with biologically relevant molecules. However, the biological role and significance of these results to the consequences of sunscreen use in humans are not known, hence extrapolation from laboratory experiments must be done with caution. (C) 2004 Elsevier Ireland Ltd. All rights reserved.