Hemodialysis continues to be the primary approach for renal replacement therapy. Vascular access (VA), particularly arteriovenous fistula or arteriovenous graft, is the preferred technique for establishing hemodialysis access due to its lower risk of infection and central venous stenosis compared to catheters. The aim of this cross-sectional investigation was to examine hemodialysis nurses’ knowledge, confidence, and practices in managing VA in Saudi Arabia. This is an online cross-sectional survey study that was conducted in Saudi Arabia between June and August 2023. This study utilized a previously validated questionnaire. Examined dialysis nurses’ knowledge, attitude, practice, and self-efficacy pertaining to VA cannulation and evaluation. Binary logistic regression analysis was used to identify predictors of better knowledge of dialysis VA. A total of 197 participants were involved in this study. Around one-third of the study participants (37.0%) reported that they have received specialized training in managing VA. Participants agreement (answered agree or strongly agree) on statements that examined attitudes toward dialysis VA cannulation and management was high and ranged between 75.0% and 93.0%. The majority of participants (97.5%) reported that they perform this assessment. Most nurses (65%) preferred the rope-ladder technique. Participants agreement on statements that examined self-efficacy on dialysis VA cannulation and management was high and ranged between 72.1% and 98.0%. The most commonly agreed upon statement was that “they have confident in performing hemodialysis access (arteriovenous fistula and arteriovenous graft) assessment before cannulation.” With 98.0% (answered agree and strongly agree). The mean knowledge score for our study sample was 6.4 (SD: 2.0) out of 12 (53.3%); which reflects marginal-level of knowledge on dialysis VA. Binary logistic regression analysis identified that nurses who have undergraduate degree are 92% more likely to be knowledgeable on dialysis VA compared to others (P < .05). Concerning VA, the level of knowledge among hemodialysis nurses working in Saudi Arabia was inadequate. Nurses who hold an undergraduate degree are presumed to have a more extensive understanding of dialysis VA. Further education programs are necessary for HD nurses to enhance their knowledge of VA, thereby optimizing their professional practices and enhancing the outcomes for their patients.