<p>Natural products are sustainable and eco-friendly sources for nanoparticle synthesis, offering unique properties and high biocompatibility. Among these, Syrian royal jelly (SRJ), rich in bioactive compounds, represents a novel and promising resource for green synthesis of gold nanoparticles (AuNPs). This study demonstrates, for the first time, the use of SRJ as bioreducing and stabilizing agent, producing uniform spherical AuNPs with significant antibacterial activity against both Gram-positive and Gram-negative bacteria. Furthermore, SRJ-AuNPs exhibited good biocompatibility with human dermal fibroblasts, indicating their potential for safe biomedical applications, including wound healing and infection prevention. By combining green synthesis with comprehensive biological evaluation, this work not only introduces a regionally sourced and multifunctional nanomaterial but also lays a foundation for future investigation into their mechanisms of action, long-term cytotoxicity, and clinical translation. Overall, these findings advance the development of eco-friendly, biologically relevant nanomaterials and open new avenues for their use in practical biomedical applications, including antibacterial coatings, wound dressings, acne treatments, and anti-aging formulations thereby, paving the way for future clinical studies.</p>