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Morphological and genotoxic analysis of PDT with fotoenticine in the treatment of gliossarcoma

Grant number: 18/15302-6
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): November 01, 2018
Effective date (End): October 31, 2019
Field of knowledge:Biological Sciences - Biophysics - Radiology and Photobiology
Principal Investigator:Juliana Ferreira Strixino
Grantee:Rainara Moreno Sanches de Almeida
Home Institution: Instituto de Pesquisa e Desenvolvimento (IP&D). Universidade do Vale do Paraíba (UNIVAP). São José dos Campos , SP, Brazil

Abstract

Gliosarcoma is a primary malignant tumor and a histopathological variant of wild type glioblastoma multiforme (GBM) isocitrate dehydrogenase (HDI), the rarest and most severe brain tumor. The current standard treatment consists of chemotherapy, radiotherapy and surgical resection of the malignant tissue. However, despite the numerous advances in the techniques, the prognosis remains unfavorable. Rarity and the high degree of specificity of the tumor necessitate the development of alternative treatments. Photodynamic therapy (PDT) is a noninvasive technique that has been highlighted as an alternative form of cancer treatment because it does not present the side effects associated with systemic treatments found in conventional methods. The reaction employs a photosensitizer (FS), the light at a specific wavelength and molecular oxygen, capable of leading to the production of reactive oxygen species. The objective of this work is to evaluate the morphology and genotoxicity of 9L / LacZ gliosarcoma cells submitted to PDT with chlorin e6 Fotoenticine. The FS will be used at concentrations of 200 ¼g / ml, 6.25 ¼g / ml and 12.5 ¼g / ml and the light source will be a LED-based device (Biopdi / IrradLed 660) consisting of 54 LEDs, with 70mW of power in the 660 nm region. For analysis of the action of the therapy, images will be performed by confocal microscopy in order to observe changes in the cytoskeleton of actin, lysosomes and mitochondria, before and after PDT. Tests will also be carried out to identify the cell death pathway after the treatment by the necrotic and apoptosis test by Annexin and Propidium Iodide, also the violet crystal tests and the comet test will be performed to evaluate the genotoxicity of the FS and images by Scanning Electron Microscopy (SEM) to observe superficial morphological characteristics.