Abstract

The human skin is exposed to various environmental factors including solar radiation and ambient air pollutants. Although, due to its physical and biological properties, the skin efficiently protects the body against the harm of environmental factors, their excessive levels and possible synergistic action may lead to harmful effects. Among particulate matter present in ambient air pollutants, PM2.5 is of particular importance for it can penetrate both disrupted and intact skin, causing adverse effects to skin tissue. Although certain components of PM2.5 can exhibit photochemical activity, only a limited amount of data regarding the interaction of PM2.5 with light and its effect on skin tissue are available. This study focused on light-induced toxicity in cultured human keratinocytes, which was mediated by PM2.5 obtained in different seasons. Dynamic Light Scattering (DLS) and Atomic Force Microscopy (AFM) were employed to determine sizes of the particles. The ability of PM2.5 to photogenerate free radicals and singlet oxygen was studied using EPR spin-trapping and time-resolved singlet oxygen phosphorescence, respectively. Solar simulator with selected filters was used as light source for cell treatment to model environmental lightning conditions. Cytotoxicity of photoexcited PM2.5 was analyzed using MTT assay, PI staining and flow cytometry, and the apoptotic pathway was further examined using Caspase-3/7 assay and RT-PCR. Iodometric assay and JC-10 assay were used to investigate damage to cell lipids and mitochondria. Light-excited PM2.5 were found to generate free radicals and singlet oxygen in season-dependent manner. HaCaT cells containing PM2.5 and irradiated with UV-Vis exhibited oxidative stress features–increased peroxidation of intracellular lipids, decrease of mitochondrial membrane potential, enhanced expression of oxidative stress related genes and apoptotic cell death. The data indicate that sunlight can significantly increase PM2.5-mediated toxicity in skin cells.

Highlights

  • We examined the impact of UVA-visible light on the toxicity of fine particulate matter (PM2.5 ) using human epidermal keratinocyte cell line (HaCaT) as a model of human epidermis

  • Many compounds commonly found in ambient particles are known to be photochemically active, we have examined the ability of PM2.5 to generate radicals after photoexcitation at different wavelengths using EPR spin-trapping

  • Previous studies showed that particulate matter could generate superoxide anion, hydroxyl radicals, and carbon-centered radicals [53,54], we have demonstrated that PM2.5, upon irradiation with

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Summary

Introduction

Skin is a natural barrier that contributes to the maintenance of the body’s homeostasis by protecting internal organs against harmful effects of various physical, chemical, and biological factors [1]. One of the physicochemical factors present in the surrounding environment that can disrupt skin homeostasis is smog [2]. The main ingredient of smog is particulate matter (PM), which can be divided into three main categories: PM10 , PM2.5 , and PM1 , representing particles of an aerodynamic diameter smaller than 10, 2.5, and 1 μm, respectively. Ambient particulate matter consists mostly of transition metal compounds (e.g., Fe(II), Cu (II)), adsorbed small reactive molecules, (e.g., environmentally persistent free radicals (EPFRs)), organic compounds (e.g., polycyclic aromatic hydrocarbons (PAHs)), minerals and soot [4,5]

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