Pain during areteriovenous fistula (AVF) cannulation remains a common problem in hemodialysis (HD) patients. This study was undertaken to assess the effect of cryotherapy on pain due to arteriovenous fistula puncture in hemodialysis patients. A convenience sample of 60 patients (30 each in experimental and control groups) who were undergoing hemodialysis by using AVF, was assessed in a randomized control trial. Hemodialysis patients who met the inclusion criteria, were randomly assigned to experimental and control groups using a randomization table. Objective and subjective pain scoring was done on two consecutive days of HD treatment (with cryotherapy for the experimental and without cryotherapy for the control group). The tools used were a questionnaire examining demographic and clinical characteristics, an observation checklist for assessing objective pain behavior, and a numerical rating scale for subjective pain assessment. Descriptive statistics were used as deemed appropriate. Chi square, two-sample and paired t-tests, the Mann Whitney test, Wilcoxon's signed rank test, the Kruskal Wallis test, and Spearman's and Pearson's correlations were used for inferential statistics. We found that the objective and subjective pain scores were found to be significantly (P = 0.001) reduced within the experimental group with the application of cryotherapy. This study highlights the need for adopting alternative therapies such as cryotherapy for effective pain management in hospital settings.