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The ability of a chitosan-based scaffold in the induction of odontogenic differentiation of dental pulp cells in culture

Grant number: 14/12017-8
Support Opportunities:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): September 01, 2014
Effective date (End): May 31, 2015
Field of knowledge:Health Sciences - Dentistry - Endodontics
Principal Investigator:Josimeri Hebling Costa
Grantee:Leopoldina de Fátima Dantas de Almeida
Supervisor: Rui L. Reis
Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
Research place: Universidade do Minho (UMinho), Portugal  
Associated to the scholarship:12/17552-3 - Blue led phototherapy on odontoblast-like and dental pulp stem cells: effect on pulp repair-related proteins production, BP.DR


The search for biomaterials that promote the regeneration of dental pulp tissue has been the subject of research in tissue engineering. Research on scaffolds, matrices formed by natural or synthetic polymers, aim to promote recovery of the tissue that suffered aggression. Chitosan, a natural polymer, has been evaluated for its bio-stimulatory and bio-reparative activities, with potential for use as scaffolds in dental pulp regeneration. Therefore, this study aims to evaluate the behavior of a human dental pulp cells in primary culture, as well as the response of a immortalized MDPC-23 cells (Mouse Dental Papilla Cells) culture, grown on a scaffold containing chitosan (50 wt%) associated the polybutylene succinate (PBS). This scaffold will be produced by means of fiber link, using the 80% deacetylated chitosan with 8.0 mm diameter and 1.0 mm thick. The human dental pulp cells used for primary culture will be obtained from dental pulp tissues removed from healthy premolars removed because of orthodontic reasons. Following the procedures of cultivation and expansion of primary culture and MDPC-23 culture, the cells will be harvested and plated at 2.5 x105 cells/scaffold density. The system will be maintained for 24 h in DMEM culture medium supplemented with 10% fetal bovine serum, at 37 ° C and 5% CO2. After that, a culture medium for osteogenic differentiation will be inserted into wells and changed every 72 h until realization of the proposed tests. The mineralized nodules production will be evaluated by Alizarin Red method; and the quantification of alkaline phosphatase will be assessed through the conversion of p-nitrophenol. The gene expression of dentin sialoprotein (DSPP) and dentin matrix protein (DMP-1) will be assessed by q-PCR. The analysis of cell morphology and scaffolds will be done using scanning electron microscopy. Tests (n = 4) will be performed at defined time-points (7, 14, 21 and 28 days), with the experimental scaffold compared to the control, containing only PBS. For all experiments, the selection of statistical tests will be based on the characteristics of the data set for each variable, such as adherence to normal curve and homoscedasticity. All statistical inferences will be made considering the significance level of 5%. (AU)

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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)
ALMEIDA, LEOPOLDINA D. F.; BABO, PEDRO S.; SILVA, CRISTIANA R.; RODRIGUES, MARCIA T.; HEBLING, JOSIMERI; REIS, RUI L.; GOMES, MANUELA E.. Hyaluronic acid hydrogels incorporating platelet lysate enhance human pulp cell proliferation and differentiation. JOURNAL OF MATERIALS SCIENCE-MATERIALS IN MEDICINE, v. 29, n. 6, . (14/12017-8)

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