Bone remodeling process evaluation during orthopedic mecanotherapy in vivo type 1 Diabetes mellitus-induced. A celular and biomolecular study

Abstract

Diabetes mellitus is associated with a several health human disorders and one of the most of its complications is the jeopardize bone formation. However, there was knowledge related to this issue, orthodontic, orthopedic treatment and celular and biomolecular evaluations. Objective: The aim of this study will be evaluate the bone remodeling process during orthopedic mecanotherapy in rats with type 1 diabetes mellitus-induced. Material and Methods: One hundred and twenty Wistar male rats will be randomly divided into eigth study groups with 15 animals each. Control Groups: normal (N), citrate buffer carrier (C), streptozotocin type 1 diabetes mellitus-induced (D) and streptozotocin type 1 diabetes mellitus-induced treated with insulintherapy (DI). Experimental Groups: normal with maxilar orthopedic expansion (NE), streptozotocin type 1 diabetes mellitus-induced with maxilar orthopedic expansion (DE) and streptozotocin type 1 diabetes mellitus-induced treated with insulintherapy associated with maxilar orthopedic expansion (DIE). The animals will be euthanized at 3, 7 and 10 days after orthopedic treatment. Changes in gens and proteins expression of osteoprotegerin (OPG), osteonectin (ONC), osteocalcin (OCC), bone sialoprotein (BSP), osteopontin (OPN), RANK, RANK-L and bone morphognetic protein (BMP) 2 and also will be evaluated the changes in body weigth, water intake and maxilla expansion suture rates will be evaluated. Real-Time PCR will be used to amplify the cDNA of OPG, RANK, RANK-L, OCC, ONC, BMP-2, BSP and OPN genes that will be analyzed on the Sequence Detection Software vl.3. These results will be confirmed by protein expression evaluations applying the Western Blotting methodologies. Data of animal weight, water intake and maxilla expansion suture rates will be analyzed statistically using Kruskal Wallis test with Dunn pos-test. The results of Real-Time PCR of each gen and the optic density values of immunoreactives bands will be clustered according to the differents study groups and the period of healing and will be analyzed using ANOVA with Student-t pos-test. A difference will be considered statistically significant when p<0.05. (AU)