Anti-MRSA and Biological Activities of Propolis Concentrations Loaded to Chitosan Nanoemulsion for Pharmaceutics Applications

Propolis is a naturally occurring substance with beneficial properties; bees produce it from various plant sources, and it is an anti-inflammatory and therapeutic resinous substance. This study aimed to enhance the biological features of propolis extract by loading it onto active film. Firstly, extraction was performed using three solvent systems, and their total phenolic, flavonoid, and antioxidant activity was measured. Propolis ethanol extract (EEP) was evaluated for phenolic fraction content and then chosen to prepare a chitosan-loaded emulsion with several concentrations. The antibacterial, anti-mycotic, and anti-mycotoxigenic properties of the extract and nanoemulsion were assessed. PPE’s cytotoxicity and nanoemulsion were evaluated using brine shrimp and cell line assays. Results indicate higher phenolic (322.57 ± 4.28 mg GAE/g DW), flavonoid (257.64 ± 5.27 mg QE/g DW), and antioxidant activity of the EEP. The phenolic fraction is distinguished by 18 phenolic acids with high p-hydroxybenzoic content (171.75 ± 1.64 µg/g) and 12 flavonoid compounds with high pinocembrin and quercetin content (695.91 ± 1.76 and 532.35 ± 1.88 µg/g, respectively). Phenolic acid derivatives (3,4-Dihydroxybenzaldehyde, 3,4-Dihydroxyphenol acetate, and di-methoxy cinnamic) are also found. Concentrations of 50, 100, 150, and 200 ng EEP loaded on chitosan nanoemulsion reflect significant antibacterial activity against pathogenic bacteria, particularly methicillin-resistant Staphylococcus aureus (MRSA) and toxigenic fungi, particularly Fusarium. Among the four EEP-loaded concentrations, the nanoemulsion with 150 ng showed outstanding features. Using a simulated medium, 150 and 200 ng of EEP-loaded chitosan nanoemulsion concentrations can stop zearalenone production in Fusarium media with complete fungi inhibition. Also, it reduced aflatoxins production in Aspergillus media, with fungal inhibition (up to 47.18%). These results recommended the EEP-chitosan application for pharmaceutics and medical use as a comprehensive wound healing agent.