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Evaluation of induced apical periodontitis in osteoporotic mice submitted or not to treatment with alendronate

Grant number: 13/18231-9
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): January 01, 2014
Effective date (End): June 30, 2017
Field of knowledge:Health Sciences - Dentistry - Pediatric Dentistry
Principal Investigator:Paulo Nelson Filho
Grantee:Priscilla Coutinho Romualdo
Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

There is little data available in the literature about the osteoporosis impact and bisphosphonates administration in apical periodontitis development and progression (Xiong et al. 2007; 2010). Considering that osteoporosis is a disease that affects worldwide the life's quality of many patients and the vast majority of these patients taking alendronate, a strong bisphosphonate, the aim of this study will be to evaluate induced apical periodontitis in osteoporotic mice submitted or not to treatment with alendronate. After appreciation of this research project by the Ethics Committee on Animal Use, School of Dentistry of Ribeirão Preto - USP, will be used 40 female mice C57BL / 6, 6-8 weeks of age. The animals will be randomly divided into 5 groups: Group I - Sham (sham surgery) (n = 5 animals), Group II - Sham and induced apical periodontitis (n = 5), Group III - animals subjected to OVX (ovariectomy) and induced apical periodontitis (n = 10), Group IV - OVX, induced apical periodontitis and treatment with sodium alendronate (ALD ) (n = 10) and Group V - OVX, induced apical periodontitis and treatment with distilled water (n = 10). The animals in groups IV and V will receive ALD or distilled water by gavage, 21 days after OVX and until the euthanasia moment. Each week the animals will be weighed to control so that a new solution will be made with the appropriate concentration of ALD according to average weight obtained. After 21 days of surgery (sham or OVX) the first lower molars of groups II, III, IV and V will be submitted to coronal access, the pulp tissues will be exposed to oral cavity with aid of spherical dental drill at low speed and the canals will remain exposed to oral microbiota for inducing apical periodontitis. After the experimental period of 21 days, it will also be performed microbiological samples of the root canals contents to the identification and the quantification of 40 species of micro-organisms in the root canals through DNA probes (checkerboard DNA-DNA hybridization) and the animals will be euthanized in a CO2 chamber. Tibias representing all groups will be removed from surgical and the proximal tibial metaphysis will be analyzed with a dualenergy X-ray densitometer to record the Bone Mineral Density (BMD) in g/cm2. The left jaws will be removed and submitted to histotechnical process. The representative sections of each group will be stained with hematoxylin and eosin (HE) and subjected to conventional optical microscopy analysis, to describe the pulp tissues and apical and periapical areas. In same time will be performed a morphometric analysis to evaluate the extension of apical periodontitis in fluorescence microscopy. Then, sequential samples will be evaluated by histoenzimology for TRAP activity to identify and counting osteoclasts. Brown and Brenn staining for the bacteria presence and location and immunohistochemistry for osteoclastogenesis markers (RANK, RANKL and OPG) and cathepsin K will be also performed. The right jaw will be removed and submitted to gene expression detection of pro-inflammatory cytokines (IL-1², IL-6, IL-10 and TNF-±) and matrix metalloproteinases (MMP-1, MMP-8 and MMP-13) by means of real-time polymerase chain reaction (RT-PCR). The results will be statistically analyzed with a significance level of 5%. (AU)

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Scientific publications (4)
(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)
SILVA, R. A. B.; SOUSA-PEREIRA, A. P.; LUCISANO, M. P.; ROMUALDO, P. C.; PAULA-SILVA, F. W. G.; CONSOLARO, A.; SILVA, L. A. B.; NELSON-FILHO, P.. Alendronate inhibits osteocyte apoptosis and inflammation via IL-6, inhibiting bone resorption in periapical lesions of ovariectomized rats. International Endodontic Journal, v. 53, n. 1, . (14/13238-8, 13/18231-9)
ROMUALDO, PRISCILLA COUTINHO; LUCISANO, MARILIA P.; PAULA-SILVA, FRANCISCO WANDERLEY G.; LEONI, GRAZIELA B.; SOUSA-NETO, MANOEL D.; SILVA, RAQUEL ASSED B.; SILVA, LEA ASSED B.; NELSON-FILHO, PAULO. Ovariectomy Exacerbates Apical Periodontitis in Rats with an Increase in Expression of Proinflammatory Cytokines and Matrix Metalloproteinases. JOURNAL OF ENDODONTICS, v. 44, n. 5, p. 780-785, . (14/13238-8, 13/18231-9)
LUCISANO, MARILIA PACIFICO; DA SILVA, RAQUEL ASSED BEZERRA; DE SOUSA PEREIRA, ANA PATRICIA; ROMUALDO, PRISCILLA COUTINHO; FERES, MAGDA; DE QUEIROZ, ALEXANDRA MUSSOLINO; NELSON-FILHO, PAULO; DA SILVA, LEA ASSED BEZERRA. Alteration of the oral microbiota may be a responsible factor, along with estrogen deficiency, by the development of larger periapical lesions. CLINICAL ORAL INVESTIGATIONS, v. 25, n. 6, . (14/13238-8, 13/18231-9)
ROMUALDO, PRISCILLA COUTINHO; FERNANDES FURLANETTO CUNHA, NAYANE BARBOSA; LEONI, GRAZIELA BIANCHI; SOUSA-NETO, MANOEL DAMIAO; CONSOLARO, ALBERTO; DE QUEIROZ, ALEXANDRA MUSSOLINO; BEZERRA DA SILVA, RAQUEL ASSED; BEZERRA DA SILVA, LEA ASSED; NELSON-FILHO, PAULO. The effect of ovariectomy and 2 antiresorptive therapeutic agents on bone response in rats: A 3-dimensional imaging analysis. ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY, v. 126, n. 3, p. 218-225, . (14/13238-8, 13/18231-9)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
ROMUALDO, Priscilla Coutinho. Evaluation of maxillary bones and apical periodontitis induced in ovariectomized rats subjected or not to treatment with bisphosphonate or cathepsin K inhibitor. 2017. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Odontologia de Ribeirão Preto (PCARP/BC) Ribeirão Preto.

Please report errors in scientific publications list by writing to: gei-bv@fapesp.br.