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Effect of the calcium glycerophosphate associated with nanoparticles of colloidal silver obtained by the 'green' synthesis about biofilms of Candida albicans and Streptococcus mutans

Grant number: 15/00825-5
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2015
Effective date (End): November 30, 2018
Field of knowledge:Health Sciences - Dentistry - Pediatric Dentistry
Cooperation agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Alberto Carlos Botazzo Delbem
Grantee:José Antonio Santos Souza
Home Institution: Faculdade de Odontologia (FOA). Universidade Estadual Paulista (UNESP). Campus de Araçatuba. Araçatuba , SP, Brazil
Associated scholarship(s):16/22039-4 - Analysis of the molecular mechanisms of toxicity and tolerance of strains of Candida albicans, Candida glabrata and Streptococcus mutans to silver nanoparticles obtained by the 'green' synthesis: approach to scale of the genome, BE.EP.DR

Abstract

The aim of this study will be to propose the synthesis and characterization of a nanocomposite based on silver nanoparticles and calcium glycerophosphate nanoparticles (Ag/calcium glycerophosphate) and evaluate its effect on planktonic and biofilms of bacteria and yeasts and assess its effect on human dental pulp cell in the presence and absence of biofilms. First, colloidal silver nanoparticles (5 nm) will be synthesized by reduction of the silver nitrate through of a "green" synthesis using a pomegranate (Punica granatum L.).Calcium glycerophosphate nanoparticles will be synthesized by grinding conventional particles of calcium glycerophosphate. These two compounds will be associated seeking a concentration of 9 wt% of phosphate. During the nanocomposite synthesis X-rays diffraction and transmission electronic microscopy will be performed. Subsequently, according to the microdilution method, it will be determined the minimum inhibitory concentration of Ag/calcium glycerophosphate capable of inhibiting the growth of C. albicans and S. mutans in the planktonic and sessile state. After, the effect of Ag/calcium glycerophosphate on preformed biofilms will be assessed through the metabolic activity of the cells. Moreover, biofilms and planktonic microorganisms will be evaluated for the production of cyclic-di-GMP using liquid chromatography coupled to tandem mass spectrometry (LC-MS) and gene expression by quantitative real-time PCR (qPCR). Finally, human dental pulp cells will be obtained from recently extracted teeth at Surgery Clinic of Araçatuba Dental School - UNESP, primary cultures (n=3) will be stablished through an explant technique and after the fourth passage the cells will be used in the assays. C. albicans and S. mutans biofilms will be formed (24 h) on inserts and used in co-culture model dental pulp cells/biofilm of both microorganisms. The effect of different concentrations and periods of treatments with nanocomposite on the human dental pulp cells, in the presence and absence of biofilms will be evaluated through 1) analysis of cell proliferation by MTT, 2) expression of alkaline phosphatase, dentin sialophosphoprotein, dentin matrix protein 1, osteocalcin and collagen type I by real-time PCR, 3) alkaline phosphatase activity assessment by thymolphthalein release by hydrolysis of thymolphthalein monophosphate, 4)detection and quantification of mineralization by alizarin red staining. Controls devoid calcium glycerophosphate and with calcium hydroxide will be included in all analyzes, and all tests will be performed in triplicate on three different occasions. Data will be analyzed to employ the most appropriate statistical test. P < 0.05 will be considered statistically significant. (AU)

Scientific publications
(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)
SANTOS SOUZA, JOSE ANTONIO; BARBOSA, DEBORA BARROS; DO AMARAL, JACKELINE GALLO; MONTEIRO, DOUGLAS ROBERTO; GORUP, LUIZ FERNANDO; DE SOUZA NETO, FRANCISCO NUNES; FERNANDES, RENAN APARECIDO; FERNANDES, GABRIELA LOPES; DE CAMARGO, EMERSON RODRIGUES; AGOSTINHO, ALESSANDRA MARCAL; BOTAZZO DELBEM, ALBERTO CARLOS. Antimicrobial Activity of Compounds Containing Silver Nanoparticles and Calcium Glycerophosphate in Combination with Tyrosol. INDIAN JOURNAL OF MICROBIOLOGY, v. 59, n. 2, p. 147-153, JUN 2019. Web of Science Citations: 0.
SOUZA, JOSE A. S.; BARBOSA, DEBORA B.; BERRETTA, ANDRESA A.; DO AMARAL, JACKELINE G.; GORUP, LUIZ F.; DE SOUZA NETO, FRANCISCO N.; FERNANDES, RENAN A.; FERNANDES, GABRIELA L.; CAMARGO, EMERSON R.; AGOSTINHO, ALESSANDRA M.; DELBEM, ALBERTO C. B. Green synthesis of silver nanoparticles combined to calcium glycerophosphate: antimicrobial and antibiofilm activities. FUTURE MICROBIOLOGY, v. 13, n. 3, p. 345-357, MAR 2018. Web of Science Citations: 1.

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