Vagner Roberto Batistela

Antibacterial photodynamic therapy for dental caries: evaluation of the photosensitizers used and light source properties.

Photodynamic therapy studies have shown promising results for inactivation of microorganisms related to dental caries. A large number of studies have used a variety of protocols, but few studies have analyzed photosensitizers and light source properties to obtain the best PDT dose response for dental caries. This study aims to discuss the photosensitizers and light source properties employed in PDT studies of dental caries. Three questions were formulated to discuss these aspects. The first involves the photosensitizer properties and their performance against Gram positive and Gram negative bacteria. The second discusses the use of light sources in accordance with the dye maximum absorbance to obtain optimal results. The third looks at the relevance of photosensitizer concentration, the possible formation of self-aggregates, and light source effectiveness. This review demonstrated that some groups of photosensitizers may be more effective against either Gram positive or negative bacteria, that the light source must be appropriate for dye maximum absorbance, and that some photosensitizers may have their absorbance modified with their concentration. For the best results of PDT against the main cariogenic bacteria (Streptococcus mutans), a variety of aspects should be taken into account, and among the analyzed photosensitizer, erythrosin seems to be the most appropriate since it acts against this Gram positive bacteria, has a hydrophilic tendency and even at low concentrations may have photodynamic effects. Considering erythrosin, the most appropriate light source should have a maximum emission intensity at a wavelength close to 530 nm, which may be achieved with low cost LEDs.

pKa determinations of xanthene derivates in aqueous solutions by multivariate analysis applied to UV-Vis spectrophotometric data.

Xanthenes form to an important class of dyes which are widely used. Most of them present three acid-base groups: two phenolic sites and one carboxylic site. Therefore, the pKa determination and the attribution of each group to the corresponding pKa value is a very important feature. Attempts to obtain reliable pKa through the potentiometry titration and the electronic absorption spectrophotometry using the first and second orders derivative failed. Due to the close pKa values allied to strong UV-Vis spectral overlap, multivariate analysis, a powerful chemometric method, is applied in this work. The determination was performed for eosin Y, erythrosin B, and bengal rose B, and also for other synthesized derivatives such as 2-(3,6-dihydroxy-9-acridinyl) benzoic acid, 2,4,5,7-tetranitrofluorescein, eosin methyl ester, and erythrosin methyl ester in water. These last two compounds (esters) permitted to attribute the pKa of the phenolic group, which is not easily recognizable for some investigated dyes. Besides the pKa determination, the chemometry allowed for estimating the electronic spectrum of some prevalent protolytic species and the substituents effects evaluation.

Evaluation of the photodynamic activity of Xanthene Dyes on Artemia salina described by chemometric approaches

The development of drugs for photodynamic therapy (PDT) is an important area of research due to their growing use in medical applications. Therefore, it is important to develop new bioassay methods for PDT photosensitizers that are inexpensive, easy to handle and highly sensitive to environmental conditions. Xanthene dyes (fluorescein, rose bengal B, erythrosine B and eosin Y) with LED light sources were investigated using Artemia salina as a bioindicator of photodynamic activity. In this study, three factors were investigated: (i) photosensitizers concentration, (ii) the LED irradiation time and (iii) the waiting time between the addition of the photosensitizers and the beginning of the irradiation. To analyze the photo-killing of A. salina, it was employed a 2³ full factorial design. The death of A. salina was related to dye structure and the interaction between the irradiation time and the photosensitizers concentration. About 60% of crustaceans death was obtained using rose bengal B, which presentes the highest quantum yield of singlet oxygen due to the number of iodide substituents in the xanthenes ring. The proposed bioassay using A. salina, xanthene dyes and LED irradiation was found suitable for quantitative PDT drug evaluation.

Influência de diferentes sistemas de solvente água-etanol sobre as propriedades físico-químicas e espectroscópicas dos compostos macrocíclicos feofitina e clorofila α

INFLUENCE OF DIFFERENT WATER-ETHANOL SOLVENT SYSTEMS ON THE SPECTROSCOPIC AND PHYSICO-CHEMICAL PROPERTIES OF THE MACROCYCLIC COMPOUNDS PHEOPHYTIN AND CHLOROPHYLL a. This work focus on the influence of solvent on the photophysical properties of chlorophyll a and pheophytin. Both compounds are related to the photosynthesis process and are considered prototypes of photosensitizers in Photodynamic Therapy. Fluorescence measurements were developed using water/ethanol mixtures at different compositions, since both solvents could be employed in biological applications. The spectroscopic properties of these compounds undergo profound changes depending on water content in the ethanol due to auto-aggregation processes. The major hydrophobicity and the lower dielectric constant of ethanol when compared with water precluded significantly the auto-aggregation process of these compounds.Keywords: chlorophyll a; pheophytin; aggregation.This work focus on the influence of solvent on the photophysical properties of chlorophyll α and pheophytin. Both compounds are related to the photosynthesis process and are considered prototypes of photosensitizers in Photodynamic Therapy. Fluorescence measurements were developed using water/ethanol mixtures at different compositions, since both solvents could be employed in biological applications. The spectroscopic properties of these compounds undergo profound changes depending on water content in the ethanol due to auto-aggregation processes. The major hydrophobicity and the lower dielectric constant of ethanol when compared with water precluded significantly the auto-aggregation process of these compounds.

Metallochlorophylls of magnesium, copper and zinc: evaluation of the influence of the first coordination sphere on their solvatochromism and aggregation properties

In this study the role of different metal centers (magnesium, zinc and copper) on the enhancement of the hydrophilic character of metallochlorophylls, was evaluated. The solvatochromism as well as the aggregation process for these compounds in water/ethanol mixtures at different volume ratios were evaluated using Fluorescence, and Resonant Light Scattering (RLS) measurements, aiming to characterize the behavior of these compounds. Independently on the studied metallochlorophyll, the presence of at least 60% of water results in a considerable increase in the fluorescence emission, probably a direct consequence of a lower aggregation of these compounds, which is confirmed by the results from RLS measurements. Additionally, the results suggest that magnesium and zinc chlorophyll should be promising phototherapeutic agents for Photodynamic Therapy.

Self-aggregation processes of 1,6-diphenyl-1,3,5-hexatriene in water/ethanol mixtures with high water percentages.

This work describes the behavior of 1,6-diphenyl-1,3,5-hexatriene (DPH) in ethanol/water mixtures. The dependence of DPH photophysical properties (absorption and fluorescence emission) on the water percentage in ethanol indicates that DPH undergoes self-aggregation processes in solvent conditions above a critical water content. Evidence such as an additional absorption band, Beers law deviation, kinetic behavior, and other experimental results obtained from temperature variation and surfactant addition demonstrated the presence of several types of DPH aggregates. Resonance light scattering measurements proved that the aggregate grew in water-rich media by a self-catalyzed process.

Chemometry in undergraduate chemistry courses: a proposal for the use of multivariate analysis in the determination of pKª

An experiment was proposed applying the Chemometric approach of Multivariate Analysis for inclusion in undergraduate Chemistry courses to promote and expand the use of this analytical-statistical tool. The experiment entails the determination of the acid dissociation constant of dyes via UV-Vis electronic spectrophotometry. The dyes used show from simple equilibrium to very complex systems involving up to four protolytic species with high spectral overlap. The Chemometric methodology was more efficient than univariate methods. For use in classes, it is up to the teacher to decide which systems should be utilized given the time constraints and laboratory conditions.

Um aparato para monitorar reações rápidas: um "stopped-flow" artesanal de baixo custo

The modern stopped-flow reaction analyzer has shown high efficiency and flexibility, which provides outstanding sample economy with a dead-time of less than 1 ms. However the cost of the equipment imposes a serious restriction to many Brazilian scientists and teachers. In this work we describe the construction of a low-cost stopped-flow system coupled to a UV-Vis spectrophotometer. The performance of the system was checked by monitoring the kinetics of two reactions: the fading of phenolphthalein in aqueous alkaline solution and the chlorophyll a demetallation in acid medium. The apparatus showed reasonable efficiency with a dead-time of 0.3 to 0.5 s. The very good results obtained in these two illustrative processes show that the system is satisfactory for determining rate constants with mean reaction times ranging from seconds to minutes.