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Item Photodynamic therapy of cationic and anionic BSA-curcumin nanoparticles on amastigotes of Leishmania braziliensis and Leishmania major and Leishmania amazonensis(Photodiagnosis and Photodynamic Therapy, Elsevier) Marcolino, Luciana Maria Cortez; Ambrósio, Jéssica Aparecida Ribeiro; Pinto, Juliana Guerra; Ferreira, Isabelle; Simioni, Andreza Ribeiro; Ferreira-Strixino, JulianaCutaneous leishmaniasis is a neglected disease prevalent in tropical countries, and conventional treatment can cause several serious side effects. Photodynamic therapy (PDT) can be considered a promising treatment alternative, as it is non-invasive therapy that has no side effects and uses accessible and low-cost substances, such as curcumin. This study evaluated the PDT response with cationic and anionic BSA nanoparticles encapsulated with curcumin in macrophages infected with L. braziliensis, L. major, and L. amazonensis. The nanoparticle system was characterized using a steady-state technique, scanning electron microscopy (SEM) study, and its biological activity was evaluated using macrophage cell lines infected with different Leishmania species. All spectroscopy measurements demonstrated that BSA curcumin (BSACur) has good photophysical properties, and confocal microscopy shows that macrophages and protozoa internalized the nanoparticles. The viability test demonstrated that at low concentrations, such as 0.1, 0.7, and 1.0 μmol. L 1, there was a decrease in cell viability after PDT application. Furthermore, a decrease in the number of parasites recovered was observed in the PDT groups. The results allowed us to conclude that curcumin loaded into BSA nanoparticles may have potential application in drug delivery systems for PDT protocols, demonstrating reduced cell viability at lower concentrations than free curcumin.Item Molecular effects of photodynamic therapy with curcumin on Leishmania major promastigotes(Parasitology Research, Springer-Verlag GmbH Germany) Marcolino, Luciana Maria Cortez; Pinto, Juliana Guerra; Ferreira, Isabelle; Godoi, Bruno Henrique; Canevari, Renata de Azevedo; Ferreira-Strixino, JulianaLeishmaniasis is a neglected disease mainly affecting low-income populations. Conventional treatment involves several side effects, is expensive, and, in addition, protozoa can develop resistance. Photodynamic therapy (PDT) is a promising alternative in treating the disease. PDT involves applying light at a specific wavelength to activate a photosensitive compound (photosensitizer, PS), to produce reactive oxygen species (ROS). Curcumin and its photochemical characteristics make it a good candidate for photodynamic therapy. Studies evaluating gene expression can help to understand the molecular events involved in the cell death caused by PDT. In the present study, RNA was extracted from promastigotes from the control and treated groups after applying PDT. RT-qPCR was performed to verify the expression of the putative ATPase beta subunit (ATPS), ATP synthase subunit A (F0F1), argininosuccinate synthase 1 (ASS), ATP-binding cassette subfamily G member 2 (ABCG2), glycoprotein 63 (GP63), superoxide dismutase (FeSODA), and glucose-6-phosphate dehydrogenase (G6PDH) genes (QR). The results suggest that PDT altered the expression of genes that participate in oxidative stress and cell death pathways, such as ATPS, FeSODA, and G6PD. The ATP-F0F1, ASS, and GP63 genes did not have their expression altered. However, it is essential to highlight that other genes may be involved in the molecular mechanisms of oxidative stress and, consequently, in the death of parasites.Item Effect of serial photodynamic therapy with curcumin on Leishmania braziliensis and Leishmania amazonensis promastigotes(CDRR Editors) Maciel, Lucas Tobias Rodrigues; Marcolino, Luciana Maria Cortez; Maciel, Fernanda Bueno Sant’Anna Pereira; Pinto, Juliana Guerra; Ferreira-Strixino, JulianaPhotodynamic Therapy (PDT) consists of using a light source and a photosensitive drug at an appropriate wavelength and molecular oxygen to trigger cell death through the production of reactive oxygen species. Because it is a localised therapy, PDT is shown to be ideal for skin diseases. American cutaneous Leishmaniasis (ACL) is a highly prevalent protozoan disease worldwide that presents different clinical evolutions and may result in ulcerations and disfiguring lesions on the skin and cartilage. This study was aimed at evaluating the effect in vitro of PDT applied serially using curcumin as a photosensitiser. For this, a concentration of 125 µg.mL-1 of curcumin was used on Leishmania braziliensis and Leishmania amazonensis strains, with a light fluence of 10 J.cm-2 and irradiance of 110 mW.cm-2. The tests done were viability analysis by trypan blue exclusion test, analysis of photosensitizer (PS) internalization by confocal microscopy and morphological alterations by May-Grunwald/Giemsa staining. We observed that there was internalisation of the PS before the first and second application of PDT, with L. braziliensis and L. amazonensis strains mortality of 92% and 82% respectively, after the second application, and induction of alterations in the structural conformation, such as cell size and non-evidence of nucleus and flagellum, demonstrating that PDT was effective. We conclude that serial PDT was effective in inducing the mortality of promastigotes forms of L. braziliensis and L. amazonensis in vitro, thus highlighting its potential for the treatment of leishmaniasis.Item Evaluation of the Photodynamic Therapy with Curcumin on L. braziliensis and L. major Amastigotes(MDPI) Pereira, André Henrique Correia; Marcolino, Luciana Maria Cortez; Pinto, Juliana Guerra; Strixino, Juliana FerreiraCutaneous leishmaniasis (CL) is a neglected disease prevalent in tropical countries with the ability to cause skin lesions. Photodynamic therapy (PDT) represents a specific and topical option for the treatment of these lesions. This study evaluated the response of macrophages infected with L. braziliensis and L. major to PDT with curcumin. Curcumin concentrations were evaluated in serial dilutions from 500.0 to 7.8 μg/mL using LED (λ = 450 ± 5 nm), with a light dose of 10 J/cm2. The Trypan blue viability test, ultrastructural analysis by scanning electron microscopy (SEM), mitochondrial polarity by Rhodamine 123 (Rho 123), curcumin internalization by confocal microscopy, and counting of the recovered parasites after the PDT treatment were performed. The lowest concentrations of curcumin (15.6 and 7.8 μg/mL) presented photodynamic inactivation. Cell destruction and internalization of curcumin in both macrophages and intracellular parasites were observed in microscopy techniques. In addition, an increase in mitochondrial membrane polarity and a decrease in the number of parasites recovered was observed in the PDT groups. This study indicates that PDT with curcumin has the potential to inactivate infected macrophages and might act as a basis for future in vivo studies using the parameters herein discussed.