Background: Hormonal dysregulation caused by renal dysfunctions and associated pathophysiological pathways, particularly hemodialysis, will naturally cause growth anomalies and even postpone puberty in adolescents. Objective: The aim of the current study was to determine the relationship between serum testosterone and estradiol levels and pubertal development in children and adolescents with end-stage renal disease (ESRD) who receive regular hemodialysis. Patients and methods: A case control study was conducted at the Pediatric Nephrology Department and Out-patient Clinics of Zagazig University Children Hospital. The study included 54 participants divided into two groups; 27 Children and adolescents with chronic hemodialysis for ESRD who was identified from the juvenile nephrology unit at the hospital and 27 healthy children and adolescents. All participants underwent a full history taking process, clinical examination, pubertal assessment using Tanner staging, routine laboratory testing, in addition to throughout the course of the trial, serum levels of girls' and boys' estradiol and total testosterone were measured. Results: Delayed puberty according to age development was found among ESRD patients by Tanner staging. Patients’ weight and height were significant lower in ESRD when compared to control group. Total testosterone in ESRD males and serum estradiol in ESRD females were significant low in comparison to controls and were associated with tanner staging. Conclusion: When compared to the general population, children with ESRD receiving regular hemodialysis experience significant impairments in pubertal growth and sexual maturation. Pubertal development is typically postponed, as demonstrated by lower serum levels of the hormones male testosterone and female estradiol.