Essential oils from medicinal and aromatic plants are increasingly recognized as a promising class of green molecules for use in crop production. In many cases, the beneficial aspects of a substance are not supported by sufficient toxicological safety testing, even though recent reports suggest that some compounds may be toxic to terrestrial or aquatic non-target species. It is, therefore, essential to investigate the possibility of adverse effects on non-target animals and humans exposed to these substances through the consumption of fruit and/or vegetables. The present study aims to examine the potential effects on yield and quality parameters and investigate the level of in vitro and in vivo toxicity of an Eco-product (EP) based on rosemary and eucalyptus essential oils, to provide a measure for safe use in the agricultural sector. The product was evaluated in lettuce crop production and indicated that one-time application of the EP formula increases yield, activating various secondary metabolism pathways of the plant to cope with oxidative stress. Cytotoxicity assays and in vivo acute oral and dermal toxicity studies suggest that the tested compound does not pose any significant health hazard, and the dissolved product can be classified in Category 5, according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).

In this study, it was demonstrated that ice-templating via freeze drying with custom-made moulds, in combination with air brushing, allows for the fabrication of poly(glycerol sebacate urethane) (PGSU) scaffolds with hierarchical multilayer microstructures to replicate various native soft tissues. The PGSU scaffolds were either monolayered but exhibited an anisotropic microstructure, or bilayered and trilayered, with each layer showing different microstructures. By using freeze drying with custom-made moulds, the ice crystals of the solvent were grown unidirectionally, and after freeze-drying, the scaffolds had an anisotropic microstructure, mimicking tissues such as tendon and skeletal muscle. The anisotropic PGSU scaffolds were also examined for their tensile strength, and a range of mechanical properties were obtained by altering the reactants’ molar ratio and polymer concentration. This is of importance, since soft tissues exhibit different mechanical properties depending on their native location and functionality. By combining freeze drying with airbrushing, scaffolds were fabricated with a thin, non-porous layer on top of the porous layers to allow three-dimensional cell co-culture for tissues such as skin and oral mucosa. These results show that fabrication techniques can be combined to produce PGSU scaffolds with tailored hierarchical microstructures and mechanical properties for multiple tissue engineering applications.