RESUMO
In the past few years, the cellular effects of ultraviolet (UV) irradiation induced in skin have become increasingly recognized. Indeed, it is now well known that UV irradiation induces structural and cellular changes in all the compartments of skin tissue. The generation of reactive oxygen species (ROS) is the first and immediate consequence of UV exposure and therefore the quantitative determination of free radical reactions in the skin during UV radiation is of primary importance for the understanding of dermatological photodamage. The RSF method (radical sun protection factor) herein presented, based on electron spin resonance spectroscopy (ESR), enables the measurement of free radical reactions in skin biopsies directly during UV radiation. The amount of free radicals varies with UV doses and can be standardized by varying UV irradiance or exposure time. The RSF method allows the determination of the protective effect of UV filters and sunscreens as well as the radical induction capacity of self-tanning agents as dihydroxyacetone (DHA). The reaction of the reducing sugars used in self-tanning products and amino acids in the skin layer (Maillard reaction) leads to the formation of Amadori products that generate free radicals during UV irradiation. Using the RSF method three different self-tanning agents were analyzed and it was found, that in DHA-treated skin more than 180% additional radicals were generated during sun exposure with respect to untreated skin. For this reason the exposure duration in the sun must be shortened when self-tanners are used and photoaging processes are accelerated.
Assuntos
Di-Hidroxiacetona/farmacologia , Radicais Livres/metabolismo , Pele/efeitos dos fármacos , Pele/efeitos da radiação , Protetores Solares/farmacologia , Raios Ultravioleta , Animais , Calibragem , Espectroscopia de Ressonância de Spin Eletrônica , Humanos , Óxidos de Nitrogênio , Pele/metabolismo , SuínosRESUMO
Free radicals/reactive oxygen species (ROS) generated in skin by UV irradiation were measured by electron spin resonance (ESR). To increase the sensitivity of measurement the short life free radicals/ROS were scavenged and accumulated by using the nitroxyl probe 3-carboxy-2,2,5,5-tetrametylpyrrolidine-1-oxyl (PCA). The spatial distribution of free radicals/ROS measured in pig skin biopsies with ESR imaging after UV irradiation corresponds to the intensity decay of irradiance in the depth of the skin. The main part of free radicals/ROS were generated by UVA (320-400 nm) so that the spatial distribution of free radicals reaches up to the lower side of the dermis. In vivo measurements on human skin were performed with a L-band ESR spectrometer and a surface coil integrating the signal intensities from all skin layers to get a sufficient signal amplitude. Using this experimental arrangement the protection of UVB and UVA/B filter against the generation of free radicals/ROS in skin were measured. The protection against ROS and the repair of damages caused by them can be realized with active antioxidants characterized by a high antioxidative power (AP). The effect of UV filter and antioxidants corresponding to their protection against free radicals/ROS in skin generated by UVAB irradiation can be quantified by the new radical sun protection factor (RSF). The RSF indicates the increase of time for staying in the sun to generate the same number of free radicals/ROS in the skin like for the unprotected skin. Regarding the amount of generated free radicals/ROS in skin as an biophysical endpoint the RSF characterizes both the protection against UVB and UVA radiation.
Assuntos
Óxidos N-Cíclicos/metabolismo , Radicais Livres/metabolismo , Pirrolidinas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Pele/efeitos da radiação , Raios Ultravioleta , Animais , Antioxidantes/farmacologia , Biópsia , Óxidos N-Cíclicos/química , Espectroscopia de Ressonância de Spin Eletrônica , Humanos , Pirrolidinas/química , Pele/efeitos dos fármacos , Marcadores de Spin , Detecção de Spin , Protetores Solares/farmacologia , SuínosRESUMO
In the last decade, naturally occurring antioxidants continue to play an important role in the food-supplement industry. The content of antioxidants in a plant depends on the species, temperature, humidity, period of growth, harvest month, part of the plant used and many other variables. Herein, we present a new method able to determine the all over antioxidative power (AP) of plant extracts or lyophilised plant parts based on the reducing activity against a stable test radical. The method is performed by ESR spectroscopy and is based on the well-known 1,1-diphenyl-2-picryl-hydrazil (DPPH) method with the major difference that both the antioxidative capacity and the antioxidative activity are used to characterise an antioxidant. The resulting antioxidative power is expressed in antioxidative units (AU), where 1AU corresponds to the activity of a 1 ppm solution of Vitamin C as a benchmark. This method allows a rapid, unexpensive and general applicable technique for the measurement of the antioxidative power of very different kinds of substances. The inclusion of the kinetic behaviour of the reducing process of the antioxidant for the determination of the AP allows the identification of the main antioxidant present in a sample. Herein, we present the application example of seeds, sprouts and adult parts of dandelion, amaranth, quinoa, fenugreek, broccoli, red clover and mugwort, where the AP method permits to characterise the plants with the highest antioxidant capacity and reaction velocity. The method permits to select active plant extracts for the food and nutrition industry.
Assuntos
Antioxidantes/farmacologia , Sequestradores de Radicais Livres/farmacologia , Radicais Livres/efeitos adversos , Extratos Vegetais/farmacologia , Ácido Ascórbico/química , Compostos de Bifenilo , Espectroscopia de Ressonância de Spin Eletrônica , Humanos , Indicadores e Reagentes/química , Picratos/química , Extratos Vegetais/química , Sementes/química , Trigonella/química , Vitamina E/químicaRESUMO
SURF_ER is a method for spectral and spatial electron spin resonance measurements on the surface of large objects which extension is only restricted by the width of the pole gap of the magnet and the homogeneity of the magnetic field and not by the cavity dimensions. The application of several techniques like SURF_ER for spectroscopic measurements, SURF_ERM for spatial scanning and SURF_ERI for spatial measurements of the depth of the surface region are discussed and represented for the skin of a human being as an example.