Alveolar repair process evaluation in rats with type 2 diabetes, induced by high-fat diet and low dose of streptozotocin: morphometric, immunohistochemistry and birefringence microscopy analysis

Abstract

Most diabetic patients are classified with type II of the disease, which is a syndrome of multiple etiology due to lack of insulin and/or the inability of insulin to properly exert its effects, resulting in the resistance of this hormone. Authors show that diabetes mellitus interferes in bone metabolism influencing tooth loss and that these losses could directly influence the type of diet and this lead to a nutritional deficiency which in turn would delay bone repair. Thus, it is necessary to understand this systemic condition in relation to cellular and structural responses of alveolar bone tissue. Aims: To analyze the dynamics of alveolar repair in non-diabetic rats and type II diabetic rats by means of immunohistochemical, morphometric and histochemical analyzes in periods of 3, 7, 14 and 42 days. Material and Methods: Fifty rats (n = 25 per group) will be used, divided into two groups: Control (CO) and Diabetic type II (DBT-2). The animals in the DBT-2 group will have diabetes induced by a hyperlipidic diet for three weeks associated with a single application of Streptozotocin (35mg.kg) dissolved in Citrate Buffer. After verification of type II diabetes, all animals will undergo exodontic surgery and will be euthanized at 3, 7, 14 and 42 days respectively. For the immunohistochemical analysis, we will use antibodies against TNF, TGF, IL6 (for inflammatory response at 3 and 7 days) TRAP, OC, WNT (for bone repair at 14 and 42 days) and for birefringence analysis, sections stained with Picrosirius- Red, will be analyzed under a polarized light microscope in order to evaluate an organization and quantity of collagen fiber bundles. For the analysis of Micro-CT will be used animals of the respective experimental groups with period of euthanasia in 42 days after exodontia. The quantitative data will be submitted to the normality test and this will indicate a more adequate statistical test (parametric vs non-parametric). It will be adopted for all tests significance level of 5%. (AU)