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Evaluation of cytokines production, gene expression and functional analysis of shed cells exposed to LTA

Grant number: 13/09152-8
Support type:Scholarships in Brazil - Master
Effective date (Start): January 01, 2014
Effective date (End): April 30, 2015
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Carlos Henrique Ribeiro Camargo
Grantee:Flávia Martins Leal
Home Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil
Associated scholarship(s):14/12261-6 - Analysis of the effect of LTA on tridimensional cell culture models for dental pulp cells and angiogenesis, BE.EP.MS

Abstract

Dental pulp inflammation is a reaction to eliminate the cause and allow the repair, after an injury such as bacterial infection. Lipoteichoic acid (LTA) is a bacterial cell wall component derived from Gram-positive bacteria, frequently found in dentinal tubules of teeth with deep caries lesions and in teeth with irreversible pulpitis, that can induce the expression of proinflammatory mediators. Cytokines are molecules involved in recruitment of inflammatory and immune cells. The inflammatory response of dental pulp stem cells by LTA is not well characterized. Dental pulp stem cells are specific mesenchymal stem cells that exist in human dental pulp tissues, which can differentiate in the presence of a tissue damage, and participate in the regeneration process. Changes in the cells can be evaluated by gene expression and functional assays. The aim of this study is to evaluate the behavior of Stem cells from Human Exfoliated Deciduous teeth (SHED) exposed to LTA, checking cytokines production, genes DSPP and DMP-1 expression, alkaline phosphatase activity and calcium deposition by alizarin red method. SHED cells will be treated with LTA, in the concentrations 1ng/mL, 10ng/mL, 100ng/mL and 1µg/mL. Gene expression of DSPP (Dentin Sialophosphoprotein) and DMP-1 (Dentin Matrix Protein-1) will be evaluated by PCR (Polymerase Chain Reaction). Alkaline phosphatase (ALP) activity will indicate mineralization process. Also, calcium deposition will be observed qualitatively by alizarin red. Cytokines IL-1ß, IL-6, TNF-± will be detected and quantified by ELISA (Enzyme Linked Immuno Sorbent Assay). Statistical analysis of ALP and ELISA will be performed by ANOVA and Tukey test (p<0,05).

Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
LEAL, Flávia Martins. Avaliação da produção de citocinas, expressão gênica e análise funcional de células SHED expostas ao LTA bacteriano. 2015. 75 f. Master's Dissertation - Universidade Estadual Paulista "Julio de Mesquita Filho" Instituto de Ciência e Tecnologia..

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