The issue of usability and accessibility in scientific journals is of crucial importance for effective knowledge communication, with a direct impact on user satisfaction. These aspects must be considered at every stage of website development, from needs analysis and planning, through design and implementation, to testing, maintenance, and further development. In the contemporary digital landscape, a discernible trend has emerged for websites to be designed by the principles of Universal Design (UD). Furthermore, tools based on artificial intelligence (AI) are playing an increasingly significant role in their development. The present study involved the execution of two independent experiments with several people overlapping in both groups. In the first study, a comparative analysis was conducted between the existing website of a scientific journal and a prototype manually developed by a designer, adhering to UD principles. In the second experiment, the existing website was compared with two prototypes generated by different AI-based tools. The evaluation of usability, accessibility, and user satisfaction was conducted utilising a combination of methodologies, including the eye-tracking technique, an author survey, and automated tools designed to assess compliance with WCAG 2.1 AA guidelines.