Diabetic cardiomyopathy is characterized by myocardial dysfunction and the production of a disorganized fibrotic matrix in the absence of coronary atherosclerosis and hypertension. We examined whether adhesion of cardiac fibroblasts to glycated extracellular matrix collagens mediates the differentiation of myofibroblasts, which may contribute to cardiac fibrosis. By microarray we found that methylglyoxal-treated collagen selectively enhanced alpha 11 integrin expression in human cardiac fibroblasts (HCF) while levels of other collagen binding integrins (alpha 1, alpha 2 and alpha 10) were unchanged. Similar increases of alpha 11 integrin expression were observed in rat cardiac fibroblasts (RCF) from streptozotocin-induced diabetic rats. In HCF plated on methyglyoxal-treated collagen and in RCF from diabetic rats, TGF-beta2 but not TGF-beta1 or TGF-beta3 were increased. Knock down of alpha 11 integrin or TGF-beta receptors with small interfering RNA abolished the increased expression of TGF-beta1 and alpha 11 integrin in cells plated on methylglyoxal-treated collagen. Our results indicate that interactions between alpha 11 integrin and TGF-beta 2 signaling may contribute to myofibroblast differentiation and the development of a fibrotic interstitium in diabetic cardiomyopathy.