Background: Associated vaccine injections pain is common in infants; pediatric nurses must employ pain management techniques in their practice. Non-pharmacological methods were reported to be more appropriate to reduce pain. Aim: The study was conducted to investigate the effect of local heat and cold application for Pentavalent vaccine injection pain in infants. Design: Quasi experimental study design was used in the present study. Method: A convenience sampling of 120 infants at El-Salam district family health center at Ismailia city throughout the period of data collection aged between 2-6 months from both sexes. Infant Information Form and FLACC Pain Scale were used to collect the data. Results: Revealed that, the mean Pain score was higher among infants who have 2 months of age, infants whose weight between 4 to less than 6 kilograms also have higher mean Pain scores in all groups. The current research demonstrates that in all groups, mean pain score were higher in female infants than in male infants. There were statistically significant variations between the mean Pain scores and the age, weight, and sex of the newborns in the three groups. Conclusion: The study concluded that there was statistically significant difference between the mean pain scores of the three studies' groups and there was statistically significant reduction in total scores of associated vaccine pain in cold application group. Recommendations: The study, emphasized on the importance of continuous training programs for pediatric nurses and parents regarding heat and cold application as method of non-pharmacological pain management.