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In vitro study of the effect of low level laser phototherapy on dentin matrix protein production by stem cells derived from human dental pulp

Grant number: 11/01943-0
Support type:Regular Research Grants
Duration: September 01, 2011 - August 31, 2013
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Marcia Martins Marques
Grantee:Marcia Martins Marques
Home Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Assoc. researchers:Cacio de Moura Netto ; Leila Soares Ferreira ; Nilton Azambuja Junior

Abstract

Dental trauma in immature permanent teeth can damage pulp vascularization and leads to a necrosis condition and cessation of the dental root formation process. The cellular therapy with the purpose of creating a new pulp tissue in theseteeth could allow the maintenance of root formation in order to finalize this process. Studies on tissueengineering to understand the mechanisms of stem cells proliferation and differentiation are very importantfor a future use on cellular therapy. The dental pulp, a soft connective tissue composed by cells andextracellular matrix (ECM), is responsible for the formation, protection and nutrition of dental structure.Several studies have shown that undifferentiated cells from this tissue can act as stem cells, capable ofdifferentiating in other cell types. There is a great potential of these dental pulp stem cells (DPSC) for thefuture treatment of pulpless immature permanent teeth with incomplete root formation. This explains thehigh interest in isolating and studying these cells. Another therapy that could be joined to cellular therapy isthe phototherapy with low intensity lasers (LPT). In fact, LPT studies have been demonstrated its capabilityof modulating cell growth, viability and differentiation, as well as the protein synthesis and secretionprocesses of several types of cells, such as fibroblasts, osteoblasts and endothelial cells. It is important tostate that the increase of cell viability becomes a major issue for stem cells, mostly when the objective oftheir culture is the implantation on human organs during cellular therapies. Some in vitro studies alreadyexplored the LPT effects on mesenquimal stem cells (derived from bone marrow and fat tissue). However,there is little knowledge about LPT effects on DPSC biology. The comprehension of how to stimulate these stem cells to proliferate and differentiate could improve the future use of these cells in dental tissues regeneration. Therefore, the objective of this study is to evaluate the LPT influence on cell growth and production of dentin extracellular matrix proteins (DSP, DPP, DMP-1, osteocalcin and type 1 collagen) of a dental pulp stem cells lineage originated from deciduous teeth (SHED-GFP cell line). For this study, it will be used cell viability analysis techniques (MTT reduction assay), immunofluorescence, Western-blot and Real time PCR. Numeric data will be statistically compared with a proper test, selected after data normality analysis. Morphologic data will be presented in a qualitative and/or illustrative manner. (AU)