Background and aim: The urgent need for breakthrough therapeutic approaches for multi-drug resistance Staphylococcus aureus (MDR) has drawn attention to potential cutting-edge tools like nanoparticles (NPs). The preparation of chitosan/alginate-encapsulated Echinacea angustifolia extract and evaluation of its effectiveness against MDR strains were the goals of this investigation. Method: Scanning electron microscopy, dynamic light scattering (DLS), and Fourier Transform Infrared spectroscopy were used to analyze the synthesized NPs. Investigations were done into the antibacterial and antibiofilm properties of E. angustifolia NPs. Also, the expression of essential genes in biofilm formation was evaluated by real-time PCR. Results: According to DLS measurements, spherical E. angustifolia NPs had a diameter of 335.3 ± 1.43 nm. The PDI was 0.681, and the entrapment effectiveness (EE%) of the E. angustifolia extract reached 83.45 %. Synthesized NPs had the highest antibacterial power. They also significantly decreased the expression of genes involved in biofilm formation. Conclusion: By releasing natural-derived medications under regulated conditions, the findings described here should help create new plant extracts with increased stability and antibacterial action, which will help minimize the usage of antibiotics.