ROS scavengers and genital skin healing in boys with hypospadias.

Introduction: Hypospadias repair is associated with high complication rates. Vascular cells from boys with hypospadias have increased reactive oxygen species (ROS) compared to controls. It is not clear if ROS affects wound healing in hypospadias.

Objectives: The aim of this study is to identify if cell migration and proliferation in genital skin is altered in hypospadias, and whether this is altered by antioxidants.

Study design: Genital skin fibroblasts (GSFs) were grown from boys undergoing hypospadias repair or routine circumcision. Cells were imaged immediately after creating a wound scratch and 48 h later, in the presence/absence of ROS scavengers, N-acetylcysteine (NAC) or Tempol. Cell migration was determined using ImageJ software. Cell proliferation was measured using a Cell Count Kit-8 (Abcam, UK).

Results: Twenty-four cases (median age (range) 1.8 (1.2, 6.3) years) and 28 controls (1.6 (1.2, 6.1) years) were recruited. Boys with hypospadias had impaired cell migration with reduced wound closure at 48 h (2.0 fold, p < 0.0001) and reduced cell proliferation (1.3 fold, p = 0.01). External Masculinisation Score was positively correlated with wound closure (r = 0.5, p < 0.0001) and cell proliferation (r = 0.3, p = 0.002). Exposure to NAC and Tempol improved wound closure (1.9 fold, p = 0.01, and 1.5 fold, p = 0.02 respectively) and cell proliferation (1.5 fold, p = 0.02 and 1.4 fold, p = 0.05 respectively).

Discussion: There is an association between wound healing and virilisation of the external genitalia in boys. ROS scavengers increase cell migration and proliferation in GSFs from boys with hypospadias.

Conclusion: Translational studies are required to confirm whether ROS scavengers may represent a therapeutic option for improving surgical outcome in boys with hypospadias.