ABSTRACT
Type 2 diabetes mellitus is increasingly becoming more prevalent worldwide together with hospital care costs from secondary complications such as bone fractures. Femoral fracture risk is higher in diabetes. Therefore, this study aimed to assess the osteometric and microarchitecture of the femur of Zucker Diabetic Sprague-Dawley (ZDSD) femur. Ten-week-old male rats (n=38) consisting of 16 control Sprague-Dawley (SD) and 22 ZDSD rats were used. The rats were terminated at 20 weeks and others at 28 weeks of age to assess age, diabetes duration effects and its severity. Bilateral femora were taken for osteometry, bone mass measurements and micro-focus X-ray computed tomography scanning to assess the trabecular number (TbN), thickness (TbTh), spaces (TbSp), bone tissue volume to total volume (BV/TV) and volume (BV). Diabetic rats had shorter (except for 20-weeks-old), lighter, narrower, and less robust bones than SD controls that wered more robust. Although cortical area was similar in all diabatic and control rats, medullary canal area was the largest in ZDSD rats. This means that the diabetic rats bones were short, light and hollow. Diabetic rats aged 20 weeks had reduced BV, BV/TV, TbN with more spacing (TbSp). In contrast, the 28 weeks old diabetic rats only showed reduced BV and TbN. Discriminant function analysis revealed, for the first time, that osteometric parameters and TbTh, TbN, and TbSp were affected by diabetes. This knowledge is valuable in the management of diabetic complications.
ABSTRACT
The endocrinology of type 2 diabetes (T2D) and its predisposing factors have been studied extensively while its skeletal effects have received negligible research despite this being a global disease. The cellular and molecular association between proximal humeral fractures and T2D has not been fully elucidated. We aimed to study bone cell quantities and immunolabel osteogenic and antiosteogenic cytokines. The study used 12-week-old rats (23 males) consisting of 8 Sprague Dawley (SD) and 15 Zucker Diabetic Sprague Dawley (ZDSD). Weekly mass measurements were taken while fasting blood glucose levels were recorded every 2 weeks with oral glucose tolerance tests conducted once every 4 weeks. Upon termination at the age of 28 weeks, humeri were fixed in 10% buffered formalin, prior to decalcification in ethylenediaminetetraacetic acid. The bone samples were then processed in ascending grades of alcohol using an automatic processor before embedding in paraffin wax. Sections were cut at 5 µm thickness in a series for Haematoxylin and Eosin stain, and immunohistochemistry was performed with the anti-tartrate-resistant acid phosphatase (TRAP), anti-alkaline phosphatase (ALP), anti-bone morphogenetic protein 3 (BMP3), anti-transforming growth factor beta 1 (TGFß1), anti-aged glycation end product (AGE) antibodies in the sequence. ZDSD rats had more adipocytes, BMP3 and AGEs expression with higher numbers of TRAP positive osteocytes and fewer ALP cells although no differences were found in TGFß1 immunopositivity. We also found that T2D increases the number of AGEs immuno-positive cells, as well as its extracellular expression, thus providing a conducive environment for the interaction of the osteogenic cytokine and its antagonist to suppress osteoblastogenesis. ZDSD groups had higher adipocyte numbers therefore increased marrow adiposity in T2D.
