Evaluation of gene expression related to the virulence factors and response to the oxidative stress of biofilms of Candida albicans submitted to antimicrobial photodynamic therapy mediated by Photoditazine® and curcumin

• Grant number: 17/12358-8
• Support type: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): October 01, 2017
• Effective date (End): July 31, 2018
• Field of knowledge: Health Sciences - Dentistry - Dental Materials
• Principal researcher: Juliana Cabrini Carmello
• Grantee: Tábata Viana de Sousa
• Home Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

The objective of this study will be to evaluate the effectiveness of antimicrobial Photodynamic Therapy (aPDT) mediated by the photosensitizers (PSs) Photodithazine (PDZ) and Curcumin (CUR) on the expression of genes responsible for the determination of virulence factors of Candida albicans biofilms such as, adhesion and biofilm formation (ALS1e HPW1 genes) and on genes responsible for the fungus response to oxidative stress (CAP1, CAT1 and SOD1 genes). In addition, the ACT 1 gene will be tested as the control gene for qPCR. For this, standardized suspensions of C. albicans ATCC 90028 will be prepared, and submitted to the adhesion phase and biofilm formation. After formation of the biofilms, the samples will be treated with 100 and 200 mg/L of PDZ or 40 and 80 ¼M of CUR for 20 minutes (pre-irradiation time), and the LED light doses at 37.5 and 50 J/cm2 for each PS concentration tested (660nm and 450nm). Additional suspensions will be only treated with the PSs or light, and the control will consist of samples that will not receive PS or light. After the proposed treatments, the 96-well plates will be placed in a sonicator for the release of the cells. After the treatments, the samples will be submitted to extraction and purification of RNA, cDNA conversion and qPCR for the evaluation of gene expression, which will be performed in the CFX96 (BioRad) equipment: step 1 (1X) 95ºC / 3 min; Step 2 (35X) 95ºC / 15 sec, 58ºC / 30 sec, 68ºC / 15 sec, for the data collection and real-time analysis; Step 3 (1X) 95 ° C / 1 min; Step 4 (1X) 55 ° C / 1 min; Step 5 (80X) 55ºC / 1 min, for analysis of the melting curve. The data obtained will be evaluated by the most appropriate statistical test. (AU)