Study of the effectiveness of antimicrobial photodynamic therapy on Candida inacti...
| Grant number: | 10/05425-1 |
| Support type: | Regular Research Grants |
| Duration: | July 01, 2010 - December 31, 2012 |
| Field of knowledge: | Health Sciences - Dentistry |
| Principal Investigator: | Ana Cláudia Pavarina |
| Grantee: | Ana Cláudia Pavarina |
| Home Institution: | Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil |
Abstract
This research project will count with the performance of three studies on which the aluminum-cholo-phthalocyanine and other derivatives associated with LED will be evaluated. On Study I two standard strains of C. albicans and C glabrata will be studied in planktonic and biofilm cultures. Aliquots of cell suspensions are transferred to a 96 wells microtiter plate with three aluminum-chloro-phthalocyanine concentrations (10, 25 and 50 mg/L) and exposed to three different light doses (18, 25.5 and 37.5 J/cm2). The biofilms will be formed on the filtering membrane surface, in which aliquots of suspensions of microorganisms are inoculated. After 48 hours, the whole is transferred to a 24 wells microtiter plate where the PDT will begin. Serial dilutions of each sample will be obtained (10-1 to 10-3), and 25 ¼L aliquots of these dilutions are plated on SDA. In Study II, two standard strains of C. albicans and C. glabrata are incubated together with cultured keratinocytes. The co-cultures will be treated in the same conditions above. After PDT, both the keratinocytes and the fungal cells will be assessed for cellular metabolism by the MTT method, cell morphology by scanning electron microscopy, type of cell death and intracellular levels of reactive oxygen species by flow cytometry. In Study III, mices with induced tongue candidosis will be submitted to PDT. After three days of inoculation, the animals are submitted to PDT, treated only with the photosensitizer, treated only with light, or receive no treatment (n=5 in each group). Afterwards, the C. albicans in the mice's tongue will be recovered by collecting them with swabs, sowing in SDA, incubating, and counting viable colonies. Moreover, analysis will be performed by macroscopic and microscopic photographs, by histology and evaluation of gene expression of cytokines and pro anti-inflammatory by PCR-RT. (AU)