Background and Aim of the Work: Inflammation is a common feature in chronic kidney disease (CKD), especially enhanced in end stage renal disease patients on hemodialysis (HD).Under inflammatory conditions, reactive oxygen species (ROS) and reactive nitrogen species (RNS) are generated from inflammatory and epithelial cells and result in DNA damage. In this study we assess DNA fragmentation in children with chronic kidney disease on regular hemodialysis sand detect its relation with some of the inflammatory markers. Material and Methods: The study included 40 children with CKD on regular hemodialysis (HD), and they were selected from the hemodialysis unit of Al-Zahraa hospital, Al-Azhar University, during the period from April 2016 to January 2018. Another group of 40 apparently healthy children, matches age and sex with patients group as a controls. Serum level of C-reactive protein (CRP), tumor necrosis factor (TNF-α) and DNA fragmentation assay in the same line with routine investigations were assessed for both groups Results: Children on regular hemodialysis have significantly higher CRP and TNF-α serum levels compared to their controls, it was (27.25±4.64 mg/L),(10.04±1.90 ng/ml) and (1.86±0.59 mg/L) (2.75±0.56 ng/ml) respectively, (P<0.001). There was a significant increase in DNA fragmentation in patients group (1.51±0.31) compared to the controls (0.54±0.21) with (P<0.001). DNA damage was present in (92%) of the study patients. There were significant positive correlations between DNA fragmentation and serum level of CRP, TNF-α, body mass index and dialysis duration. Conclusion: Almost children on hemodialysis are exposed to DNA damage with significant association with inflammatory markers, duration of hemodialysis and BMI.