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Study of angiogenesis and temporal expression of VEGF and its receptor VEGFR-1/Flt-1 and VEGFR-2/Flk-1 during alveolar bone repair and normal therapeutic use of a non-steroidal anti-inflammatory selective for COX-2 in rats

Grant number: 09/11333-5
Support type:Scholarships in Brazil - Master
Effective date (Start): March 01, 2010
Effective date (End): September 30, 2011
Field of knowledge:Biological Sciences - Morphology - Histology
Principal researcher:Rumio Taga
Grantee:Ricardo Vinicius Nunes Arantes
Home Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil

Abstract

The formation and development of an active microvasculature is an essential stage for the remodeling and bone repair. Among the main factors related to the control of angiogenesis highlights the vascular endothelial growth factor (VEGF) which currently, has been proposed as a signaling molecule in the communication between endothelial cells and osteoblasts. Furthermore, the prostaglandins (PGs) from arachidonic acid by catalysis of cyclooxygenase (COX) in response to the inflammatory process, injury and mechanical stress regulate the angiogenesis and bone formation. The PGs stimulate osteoblasts and fibroblasts, to express the VEGF which in turn has paracrine effects on endothelial cells and the precursors of osteoblasts. The therapeutic effects of non-steroidal anti-inflammatory drugs (NSAIDs), used to control the swelling, fever, and pain, are related to its inhibitory effect of COX-2. Many reports in the literature have demonstrated the negative effect of chronic administration of NSAIDs in the process of bone repair. Already, in dentistry, the NSAIDS are widely used in therapy for control of acute pain after tooth extraction and its effect on angiogenesis and ossification has been little studied. Therefore, the current work we propose to evaluate morphometrically the process of alveolar bone healing after tooth extraction correlating it with the temporal expression of VEGF and its receptors VEGFR/Flt-1 and VEGFR2/Flk-1 with or without the use of therapeutic an NSAID. Thus, the extraction of upper left incisor is performed in 180 rats (Rattus norvegicus) Wistar, male, with 60 days of age, divided into two groups: 1) control group (n = 90) - the animals will receive daily intraperitoneal injection of 0.1 ml of 0.9% saline solution daily for 7 days, and 2) treated group (n = 90) -the animals receive a daily injection intraperiotoneal 3mg/kg body mass Meloxican in saline 0, 9% daily for 7 days. After experimental periods of 3, 7, 10, 14, 21 and 30 days, the cells will be carefully collected, of which 5 are well established in the 10% formalin in phosphate buffer and processed histologically for morphometric analysis and immunohistochemical , 5 cells are crushed and immersed in TRIzol ® to evaluate the expression of the VEGF RNAms and their receptors by Real Time-PCR and 5 cells are crushed and frozen to -80 º C and subsequently for evaluation of VEGF protein and its receptors by western blotting. In the analysis will be assessed: a) morphometrically the volume density of blood vessels and the neoformed bone tissue and its correlation with tissue expression of VEGF and its receptors, b) the temporal and spatial cell types immunomarked against VEGF and its receptors; c) the kinetics of the number of cells of the alveolar imunomarcadas/mm2 against VEGF and its receptors and the expression of their RNAms by Real-Time PCR, and d) possible changes in the pattern of protein expression of VEGF and its receptors by Western blotting. The comparison between the groups will test our hypothesis that the use of Meloxicam in the treatment of acute postoperative pain can negatively influence the COX-2, delaying the bone repair (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)
NUNES ARANTES, RICARDO VINICIUS; CESTARI, TANIA MARY; VISCELLI, BRUNO ALVARES; DIONISIO, THIAGO JOSE; GARLET, GUSTAVO POMPERMAIER; SANTOS, CARLOS FERREIRA; DE ASSIS, GERSON FRANCISCO; TAGA, RUMIO. Meloxicam Temporally Inhibits the Expression of Vascular Endothelial Growth Factor Receptor (VEGFR)-1 and VEGFR-2 During Alveolar Bone Repair in Rats. Journal of Periodontology, v. 86, n. 1, p. 162-172, . (09/11333-5)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)

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