This thesis intends to utilize biomechanics to study the endothelial biomechanical response in a static hyperglycemic microenvironment. Hyperglycemia is a diabetic condition with abnormally high levels of glucose in the bloodstream. The effects of hyperglycemia over time lead to vascular complications resulting in patients being more prone to cardiovascular diseases. Current studies have focused on the molecular mechanisms affected by hyperglycemia; however, the mechanical mechanisms by which hyperglycemia causes vascular structural and functional changes are understudied. Therefore, to study the effects of hyperglycemia in the endothelium, Human Umbilical Vein Endothelial Cells (HUVEC) were cultured under three glucose conditions: normal glucose (4 mmol/l D-glucose), high glucose (30 mmol/l D-glucose), and an osmotic control (4 mmol/l D-glucose + 26 mmol/l D-mannitol). To evaluate the biomechanical response, we used traction force microscopy and monolayer stress microscopy to measure the cell-substrate tractions and cell-cell intercellular stresses. For the RMS tractions, HUVEC monolayers exposed to high glucose decreased by 10%, while the osmotic control decreased by 17% compared to the normal glucose. HUVEC monolayers exposed to high glucose produced average normal stresses that were 53% lower than monolayers exposed to normal glucose, while the osmotic control decreased by 51% compared to the normal glucose. For the maximum shear stresses, HUVEC monolayers exposed to high glucose decreased by 20%, while the osmotic control decreased by 14% compared to the normal glucose. To conclude this study, we report that hyperglycemia lowers the biomechanical response in the endothelium compared to normal glucose conditions. These results will contribute to understanding the specific role hyperglycemia has on endothelial mechanics and its role in the progression and development of cardiovascular diseases in diabetic patients.