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Evaluation of the effectiveness of antimicrobial photodynamic therapy in the inactivation of Candida species and cellular damage caused to skin cells: in vitro and in vivo evaluation of the aluminum-chloro-phtalocyanine and other derivatives

Grant number: 10/05425-1
Support Opportunities:Regular Research Grants
Start date: July 01, 2010
End date: December 31, 2012
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Ana Cláudia Pavarina
Grantee:Ana Cláudia Pavarina
Host 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)

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Scientific publications (4)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DIAS RIBEIRO, ANA PAULA; ANDRADE, MARIANA CARVALHO; BAGNATO, VANDERLEI SALVADOR; VERGANI, CARLOS EDUARDO; PRIMO, FERNANDO LUCAS; TEDESCO, ANTNIO CLAUDIO; PAVARINA, ANA CLAUDIA. Antimicrobial photodynamic therapy against pathogenic bacterial suspensions and biofilms using chloro-aluminum phthalocyanine encapsulated in nanoemulsions. Lasers in Medical Science, v. 30, n. 2, p. 549-559, . (09/17975-9, 10/05425-1)
SANITA, PAULA VOLPATO; PAVARINA, ANA CLAUDIA; DOVIGO, LIVIA NORDI; RIBEIRO, ANA PAULA DIAS; ANDRADE, MARIANA CARVALHO; DE OLIVEIRA MIMA, EWERTON GARCIA. Curcumin-mediated anti-microbial photodynamic therapy against Candida dubliniensis biofilms. Lasers in Medical Science, v. 33, n. 4, p. 709-717, . (09/17975-9, 13/07276-1, 10/05425-1)
DIAS RIBEIRO, ANA PAULA; PAVARINA, ANA CLAUDIA; DOVIGO, LIVIA NORDI; BRUNETTI, IGUATEMY LOURENCO; BAGNATO, VANDERLEI SALVADOR; VERGANI, CARLOS EDUARDO; DE SOUZA COSTA, CARLOS ALBERTO. Phototoxic effect of curcumin on methicillin-resistant Staphylococcus aureus and L929 fibroblasts. Lasers in Medical Science, v. 28, n. 2, p. 391-398, . (08/03994-9, 10/05425-1)
DIAS RIBEIRO, ANA PAULA; ANDRADE, MARIANA CARVALHO; DA SILVA, JULHIANY DE FATIMA; JORGE, JANAINA HABIB; PRIMO, FERNANDO LUCAS; TEDESCO, ANTONIO CLAUDIO; PAVARINA, ANA CLAUDIA. Photodynamic Inactivation of Planktonic Cultures and Biofilms of Candida albicans Mediated by Aluminum-Chloride-Phthalocyanine Entrapped in Nanoemulsions. Photochemistry and Photobiology, v. 89, n. 1, p. 111-119, . (08/03994-9, 10/05425-1)