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Effect of different irrigation protocols on the attachment and differentiation of stem cells from the apical papilla

Grant number: 15/09958-8
Support type:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): February 15, 2016
Effective date (End): February 14, 2017
Field of knowledge:Health Sciences - Dentistry - Endodontics
Principal researcher:Clovis Monteiro Bramante
Grantee:Talita Tartari
Supervisor abroad: Renato Menezes Silva
Home Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
Research place: University of Texas Health Science Center at Houston (UTHealth), United States  
Associated to the scholarship:13/19789-3 - Effects of different irrigation regimes in physico-chemical properties of dentin and consequences of changesin the adhesion of microorganisms and AH Plus sealer, BP.DR


In regenerative endodontics procedures (REPs) the root canal system disinfection with irrigation solutions and medicaments is crucial. However, irrigants can alter the dentin which may affect stem cells attachment and survival. Therefore, the aims of this study are to investigate whether the dentin treatment with different irrigation protocols affect the attachment and differentiation of stem cells from the apical papillae (SCAPs) and to analyze if the phenotypical changes of SCAPs are related to the biochemical components from the dentin surface. A characterized SCAP line will be used in all experiments. The SCAPs will be selected, cultured and seeded on dentin discs treated with different endodontic irrigation protocols. After 3 days of culture, cell attachment will be evaluated by cell density, fibronectin 1 gene expression levels using quantitative real-time polymerase chain reaction (qRT-PCR), and scanning electron microscopy. After 21 days of culture, a PCR Array will be used to verify the expression of 84 genes related to the identification, growth and differentiation of stem cells, and calcification will be investigated using alizarin red S staining. All experiments will be repeated at least in triplicate under 3 independent conditions. Data are going to be presented as the median and minimum and maximum values. The nonparametric tests of Mann-Whitney U and Kruskal-Wallis will be performed to compare the cell density and attachment and gene expressions levels. The Multiple Linear Regression test will be applied to determine if there is correlation between the cell density and attachment and phenotypical changes of SCAPs with the roughness, wettability and composition of the dentin, data collected during the execution the PhD scholarship in Brazil. All hypotheses will be tested with a 95% confidence level.

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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)
TARTARI, T.; WICHNIESKI, C.; BACHMANN, L.; JAFELICCI, JR., M.; SILVA, R. M.; LETRA, A.; VAN DER HOEVEN, R.; DUARTE, M. A. H.; BRAMANTE, C. M. Effect of the combination of several irrigants on dentine surface properties, adsorption of chlorhexidine and adhesion of microorganisms to dentine. International Endodontic Journal, v. 51, n. 12, p. 1420-1433, DEC 2018. Web of Science Citations: 0.

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