Elemental Analysis of Rat's Femoral Neck with Experimental Diabetes by means of Scanning Electron Microscopy

Diabetes mellitus (DM) is a group of metabolic disorders, whose main characteristic is the hyperglycemia. The duration of the hyperglycemia as well as its severity, are the most important factors that establish its presence by intermediate or prolonged periods. Although, pathophysiological changes that accompany the DM are found in all body, there are specific clinical manifestations such as gradual vision loss, renal disease, and susceptibility to cardiovascular diseases. In addition, the skeleton is affected to a significant level, where a major complication is osteoporotic bone fracture. Both patients with type 1 (T1DM) and type 2 (T2DM) diabetes mellitus have a high risk to experience fracture (Vestergaard et al 2009), when its condition is compared to health individuals. Reported data suggest that patients with T1DM show a decrease in bone mineral density (BMD), which is attributed to the decrease of the bone formation during growth in children and adolescents (Hamann et al 2012). In contrast, adults with T2DM, BMD values are normal or slightly elevated; however, the risk of a fracture is high (Janghorbani et al 2007). This advises that these patients may have a poor bone quality, which cannot be estimated in conventional densitometry studies. In this paper, we use a model of experimental diabetes (ED) to evaluate macroscopically the femur; representing the part of the skeleton, in which fractures caused by the weaking of the skeleton occur more frequently. In this study, female Wistar rats were used, with 8 to 12 weeks old, and weighing 200 to 250 g. The slaughter and handling of animals was made considering the specifications outlined in the Official Mexican Standard NOM-062-ZOO-1999. Before the induction of ED, the rats were fasted for 4 hours, after this time, alloxan monohydrate (Sigma-Aldrich) was administrated intraperitoneally (180 mg/kg). Animals with blood glucose values > 150 mg/dL were considered diabetic. Rats were maintained under these conditions for 12 months before sacrifice. Prior to sacrifice, body weight and blood glucose level were recorded. The rats were sacrificed by cervical dislocation and immediately skeleton dissection was performed. To prevent damage to the bone by a chemical treatment, the bones were cleaned using beetles, Coleoptera order and Dermestidae family. Our results show that the glucose level increased significantly from 53 ± 8 in the group for non-diabetic rats to values of *234 ± 39 for the group of diabetic. *p < 0.05 student’s t-test, n = 3 ♀, two weeks after induction and kept under these conditions for 12 months. This glucose level in rats corresponds to the clinical condition of non-controlled diabetic patients. Femur macroscopic parameters such as weight, length, and diameter of the diaphysis decreased  40%,  10%, ~ 7% respectively; while the diameter of the upper and lower epiphysis increased  2%,  10%. Analysis by Scanning Electron Microscopy (SEM) shows that the group of rats with ED increased femoral neck diameter  30% resulting in widening and deformation of the head of the bone (see Figure 1A and 1B).