Abstract: Three-point bending test is the most common mechanical test used for quantifying the biomechanical quality of bone tissue and bone healing in small animals. However, there is a lack of standardization for evaluation of bone repair by cortical perforation. The aim of this study was to determine the influence of bone defect position in the proximal metaphysis of rat tibias during load application and different span configuration on the three-point bending test outcomes. Cortical defects with 1.6 mm diameter were created at a standardized location on the medial surface of 60 tibias of male Wistar rats. The animals were euthanized 7 days after surgery. Five specimens were used to create 3D models for finite element analysis using high-resolution micro-CT images. Two spans (6 and 10mm) and three positions of the bone defect in relation to the load application (upward, frontal and downward) were evaluated experimentally (n = 10) and in finite element analysis (n = 5). Maxim um load (N) and stiffness (N/mm) were statistically analyzed with 2-way ANOVA and Tukey test (α = 0.05). The results demonstrated that span and orientation of the bone defect significantly influenced the fracture pattern, stress distribution and force versus displacement relation. Therefore, reliable outcome can be achieved creating the bone defect at 8 mm from the extremity of the proximal epiphysis; placing a 10 mm distance span and downward facing defect position to allow a better distribution of stress and more fracture patterns that reached the bone defect target area with less intra-group variability.
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