Bone healing in critical-size defects treated with bone marrow aspirate. Histological, histometric and immunoistochemical study in rat calvarial

Abstract

The purpose of this study is evaluate histomorphometrically and immunoistochemically the influence of Bone Marrow Aspirate (BMA)on healing process of critical-size defects (CSD) surgically created in rat calvarial. 40 rats will be divided in 2 groups: Group C (control) and Group BMA. A 5-mm diameter CSD will be created in the calvarium of each animal. In Group C, the defect was filled with blood clot only. In Group BMA, the defect was filled with BMA. The animals were euthanized at 15 or 30 days post-operatively. The area of the original surgical defect and the surrounding tissues will be removed en bloc. The specimes will be processed and embedded in paraffin. Serial sections 6 µm-thick will be cut in a longitudinal direction to the sagittal suture. Some sections will be stained with hematoxylin and eosin for histomorphometric analysis. The amount of new bone formation will be calculated as a percentage of the total area of the original defect. Other sections will be submitted to immunohistochemical reactions for detection of proliferating cell nuclear antigen (PCNA), runt-related transcription factor - 2 (RUNX-2) and vascular endothelial growth factor (VEGF). The results will be analyzed by light microscopy. The data will be submitted to statistical analysis. (AU)