Zein is renewable plant protein with valuable film-forming properties that can be used as a packaging material. It is known that the addition of natural cross-linkers can enhance a film’s tensile properties. In this study, we aimed to prepare antimicrobial zein-based films enriched with monolaurin, eugenol, oregano, and thyme essential oil. Films were prepared using the solvent casting technique from ethanol solution. Their physicochemical properties were investigated using structural, morphological, and thermal techniques. Polar and dispersive components were analyzed using two models to evaluate the effects on the surface free energy values. The antimicrobial activity was proven using a disk diffusion method and the suppression of bacterial growth was confirmed via a growth kinetics study with the Gompertz function. The films’ morphological characteristics led to systems with uniform distribution of essential oils or eugenol droplets combined with a flat-plated structure of monolaurin. A unique combination of polyphenolic eugenol and amphiphilic monoglyceride provided highly stretchable films with enhanced barrier properties and efficiency against Gram-positive and Gram-negative bacteria, yeasts, and molds. The prepared zein-based films with tunable surface properties represent an alternative to non-renewable resources with a potential application as active packaging materials.

Biofilm is a structured community of microorganisms adhering to surfaces of various polymeric materials used in food packaging. Microbes in the biofilm may affect food quality. However, the presence of biofilm can ensure biodegradation of discarded packaging. This work aims to evaluate a biofilm formation on the selected biodegradable polymer films: poly (lactic acid) (PLA), poly (butylene adipate-co-terephthalate) (PBAT), and poly (butylene succinate) (PBS) by selected bacterial strains; collection strains of Escherichiacoli, Staphylococcusaureus; and Bacillus pumilus, Bacillussubtilis, Bacillustequilensis, and Stenotrophomonasmaltophilia isolated from dairy products. Three different methods for biofilm evaluation were performed: the Christensen method, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and fluorescence microscopy. High biofilm formation was confirmed on the control PBS film, whereas low biofilm formation ability was observed on the PLA polymer sample. Furthermore, the films with incorporated antimicrobial compounds (thymol or eugenol) were also prepared. Antimicrobial activity and also reduction in biofilm formation on enriched polymer films were determined. Therefore, they were all proved to be antimicrobial and effective in reducing biofilm formation. These films can be used to prepare novel active food packaging for the dairy industry to prevent biofilm formation and enhance food quality and safety in the future.

Poloxamers (P184, P188, and P407) have been investigated as the carrier system for eugenol or thymol. A synergic effect of mixed Poloxamers was proved by enhanced micellar parameters, with a lower critical micelle concentration (about 0.06 mM) and the highest surface adsorption of 9 × 10^-7 mol m^-2 for P188/P407. Dynamic light scattering revealed a decrease in micellar size after loading with biomolecules. Three mathematical models were applied to study the release kinetics, of which Korsmeyer-Peppas was the best fitted model. Higher relative release was observed for Poloxamer/eugenol samples, up to a value of 0.8. Poloxamer micelles with thymol were highly influential in bacterial reduction. Single P407/eugenol micelles proved to be bacteriostatic for up to 6 h for S. aureus or up to 48 h for E. coli. Mixed micelles were confirmed to have prolonged bacteriostatic activity for up to 72 h against both bacteria. This trend was also proven by the modified Gompertz model. An optimized P188/P407/eugenol micelle was successfully used as a model system for release study with a particle size of less than 30 nm and high encapsulation efficiency surpassing 90%. The developed mixed micelles were proved to have antibiofilm activity, and thus they provide an innovative approach for controlled release with potential in topical applications.