Background :Patients with end-stage renal disease undergoing hemodialysis are repeatedly exposed to stress and pain from approximately 300 punctures per year to their arteriovenous fistula (AVF). Aim of the study is to investigate effect of cryotherapy on pain intensity at puncture sites of arteriovenous fistula among adult patients undergoing hemodialysis. Subjects and method: Quasi experimental design (pre-post test) was utilized to perform the study. Thirty adult patients undergoing hemodialysis in hemodialysis unit in Tanta University Hospital were enrolled in the study. Patients included in the study were considered at first time as a control group where no cryotherapy used and at second time for three consecutive days of dialysis the same patients considered as a study group when cryotherapy were done. Two tools utilized for the study included: structured interview questionnaire and subjective pain assessment tool (numerical pain rating scale). The results showed that pain from arterial puncture needle was prevalent in 80 % of patients. More than half of patient (57. %) prefers change the insertion site of needle. There was statistical significant difference before intervention and after intervention by cryotherapy. There was also significant difference in the three consecutive days of intervention. Conclusion: the study concluded that cryotherapy is effective in reducing AV fistula puncture pain of hemodialysis patients. Recommendation: Hemodialysis units should involve cryotherapy for managing needle puncture pain in the routine care for hemodialysis patients.