OBJECTIVE: Obese children have severe endothelial dysfunction as measured by flow-mediated dilation (FMD). We have shown that folic acid normalizes endothelial function in children with type 1 diabetes who have a similar degree of endothelial dysfunction but lower total plasma homocyst(e)ine (tHcy) and higher folate status. Our aim was to evaluate, for the first time, the effect of folate supplementation on endothelial dysfunction in obese children. RESEARCH DESIGN AND METHODS: A total of 53 obese subjects (26 male, mean +/- SD age 13.3 +/- 2.2 years, and BMI Z score 2.29 +/- 0.25) participated in a randomized, double-blind, placebo-controlled, parallel trial of oral folic acid (5 mg/day) or placebo for 8 weeks. Before and after the intervention, we assessed endothelial function (FMD), smooth muscle function (glyceryl trinitrate-induced dilatation [GTN]), high-sensitivity C-reactive protein (hsCRP), tHcy, serum folate, red cell folate (RCF), and lipids. RESULTS: There were no group differences at baseline. FMD did not change with the intervention (folic acid group pre- and postintervention: 6.42 +/- 5.03 and 6.56 +/- 4.79%, respectively, vs. placebo group: 5.17 +/- 3.54 and 5.79 +/- 4.26%, respectively; P = 0.6). Folate supplementation increased serum folate and RCF by 18.4 nmol/l (P < 0.001) and 240.1 nmol/l (P < 0.001), respectively, and decreased tHcy by 0.95 microl (P = 0.008). The intervention did not change GTN, hsCRP, or lipids. CONCLUSIONS: Folic acid supplementation does not improve endothelial function in obese children without diabetes despite increasing folate status and reducing tHcy. This is in contrast to the response to folate in children with type 1 diabetes.