Background: The consumption of advanced glycation end products (AGEs) has increased because of modern food processing and has been linked to the development of type 2 diabetes in rodents. Objective: We determined whether changing dietary AGE intake could modulate insulin sensitivity and secretion in healthy, overweight individuals. Design: We performed a double-blind, randomized, crossover trial of diets in 20 participants [6 women and 14 men; mean 6 SD body mass index (in kg/m 2 ): 29.8 6 3.7]. Isoenergetic-and macronutrientmatched diets that were high or low in AGE content were alternately consumed for 2 wk and separated by a 4-wk washout period. At the beginning and end of each dietary period, a hyperinsulinemiceuglycemic clamp and an intravenous glucose tolerance test were performed. Dietary, plasma and urinary AGEs N V -(carboxymethyl) lysine (CML), N V -(carboxyethyl)lysin (CEL), and methylglyoxalderived hydroimadazolidine (MG-H1) were measured with the use of mass spectrometry. Results: Participants consumed less CML, CEL, and MG-H1 during the low-AGE dietary period than during the high-AGE period (all P , 0.05), which was confirmed by changes in urinary AGE excretion. There was an overall difference in insulin sensitivity of 22.1 mg $ kg 21 $ min 21 between diets (P = 0.001). Insulin sensitivity increased by 1.3 mg $ kg 21 $ min 21 after the low-AGE diet (P = 0.004), whereas it showed a tendency to decrease by 0.8 mg $ kg 21 $ min 21 after the high-AGE diet (P = 0.086). There was no difference in body weight or insulin secretion between diets (P = NS). Conclusions: A diet that is low in AGEs may reduce the risk of type 2 diabetes by increasing insulin sensitivity. Hence, a restriction in dietary AGE content may be an effective strategy to decrease diabetes and cardiovascular disease risks in overweight individuals. This trial was registered at clinicaltrials.gov as NCT00422253.