Objective <p>This study aims to evaluate the scientific accuracy, content quality and reliability of HPV vaccination videos published on YouTube using the Global Quality Score (GQS), DISCERN, and the Journal of the American Medical Association (JAMA) scoring systems.</p> Method <p>This cross-sectional study used the video-sharing website YouTube. The search terms ‘HPV vaccine’ and ‘HPV vaccination’ were used. All videos were evaluated by two physicians, and 113 videos were included in the study. The quality and reliability of the information in the videos searched on April 22, 2025 were evaluated using GQS, DISCERN, and JAMA scales.</p> Findings <p>Of the 113 videos analyzed, the vast majority (90.3%) reflected a positive attitude towards the HPV vaccine. When evaluated for reliability using DISCERN scores, 24.8% of the videos were deemed to have very poor or poor reliability, 25.7% were considered to have good reliability, and 20.4% were rated as having excellent reliability. Based on GQS scores, 36.3% of the videos were deemed of high quality. Overall, videos from healthcare organizations were found to be more reliable and of higher quality than those from non-experts and media/news sources. Videos from expert channels received higher engagement. DISCERN and JAMA reliability scores, along with GQS quality scores, were significantly higher in expert-sourced videos. A positive correlation was found between video length, quality, and reliability scores.</p> Conclusion <p>It was found that videos from expert sources have higher quality standards and that healthcare professionals play an important role on digital platforms. Health-related information must come from reliable, and scientifically based sources. Increasing the accessibility of videos produced by healthcare experts is critical in preventing the spread of misinformation and myths.</p>