<p>The increasing demand for sustainable and cost-effective pavement technologies has led to a growing focus on maximizing the Reclaimed Asphalt Pavement (RAP) utilization in asphalt mixtures. The incorporation of high-RAP content in Hot Mix Asphalt mixtures presents challenges related to aged binder stiffness, reduced workability, and the development of non-homogeneous asphalt mixtures. To address these issues, bio-rejuvenators have emerged as a promising solution, effectively restoring the aged binder properties and enhancing overall performance. This review systematically synthesizes current knowledge on the concept, types, and requirements of bio‑rejuvenators; optimum dosage determination; rejuvenation efficiency; and laboratory and field performance of bio‑rejuvenated high‑RAP mixtures. A structured four‑step methodology, supported by bibliometric analysis, was employed to identify research trends, knowledge gaps, and thematic clusters. The consolidated findings from the laboratory and field evaluations indicate positive outcomes, demonstrating the potential for increased RAP utilization without compromising performance. Based on this synthesis, the review proposes specific guidelines for advancing bio‑rejuvenator technology and implementation. These include selection criteria for bio‑rejuvenator sources, standardized dosage determination methods, strategies for enhancing long‑term durability, integration of anti‑aging additives, and environmental impact assessment. This review aims to support researchers, pavement engineers, policymakers, and industry practitioners in optimizing high-RAP mixtures through sustainable and performance-driven approaches. By combining a rigorous methodological framework with actionable recommendations, this study contributes to advancing scientific understanding of bio-rejuvenators and directs future research toward the development of standardized, field-validated practices.</p>