Background: Non-pharmacologic interventions have been recommended to relief pain in neonates during procedures related to the acute pain of mild to moderate intensity. They have proven the efficiency and present low risks to neonates, as well as low operating cost concerning intensive care. Aim of the study was to assess the effect of an educational program on enhancing pediatric nurses' performance regarding selected non-pharmacological techniques to alleviate pain in neonates. Research design: Quasi-experimental research design was utilized in the present study. The study was carried out at the neonatal intensive care unit affiliated to Zagazig University Children's Hospital. All available nurses (no=44), were included in this study. Two tools were used for data collection 1) Structured interview questionnaire sheet, and 2) Observational checklist. Results: It was revealed that the highest percentage of nurses and all of them had an unsatisfactory and inadequate level of knowledge and practice regarding selected non-pharmacological techniques to alleviate pain in neonates before the implementation of the program (79.5% &100%, respectively). Meanwhile, after the implementation of the program, about two third and slightly less than half of nurses had satisfactory and adequate levels of knowledge and practice (68.2% & 47. 7%, respectively), with a statistically significance difference (p=0.000). Conclusion: Implementing the educational program had a significant positive effect on improving pediatric nurses' performance regarding selected non-pharmacological techniques to alleviate pain in neonates. Recommendations: Encouraging more periodical continuing in service education for pediatric nurses that empowering more successive utilization of non-pharmacological methods in clinical care with the need of follow up.