Background <p>Artificial intelligence is expected to revolutionize healthcare delivery, transform the role of nurses, and enhance patient outcomes. The knowledge-based artificial intelligence chatbot has been given prominence as interactive learning aids that can provide individual assistance and support educators through various means. This study was conducted to evaluate the effectiveness of integrating a knowledge-based AI chatbot system into nursing training programs in Egypt and Saudi Arabia.</p> Methods <p>A quasi-experimental design was used, and data were collected from a purposive sample of 146 nurses, 73 nurses from Egypt, and 73 nurses from Saudi Arabia via Google Forms. The tools applied within the study included a structured questionnaire for nurses, demographic data, nurses’ knowledge of artificial intelligence in nursing, perceptions of chatbot applications in nursing education, and opinions on the use of artificial intelligence in providing nursing care.</p> Results <p>There were highly statistically significant improvements in nurses’ knowledge after using a knowledge-based artificial intelligence chatbot in nursing training programs (<i>p</i> &lt; 0.05). The most notable knowledge gains occurred in understanding AI’s role in enhancing the function of nurses in the future (Egypt: 46.6–80.3%; Saudi Arabia: 34.2–43.8%) and its applications in patient care improvement (Egypt: 39.7–61.7%; Saudi Arabia: 39.7–49.3%). Additionally, both countries demonstrated a significant improvement in nurses’ perceptions of artificial intelligence after the intervention (<i>p</i> &lt; 0.05). Post-intervention scores showed substantial increases across all perception statements, with most items exceeding 90% agreement.</p> Conclusion <p>Integrating a knowledge-based artificial intelligence chatbot into nursing training programs increased nurses’ knowledge and perceptions in Egypt and Saudi Arabia, highlighting its potential to enhance nursing education and practice.</p> Clinical trial number <p>Not Applicable.</p>