A systematic study of fabricating nanoparticles (NPs) by cost-effective polymer deposition/solvent displacement (nanoprecipitation) method has been carried out. Five amino acid based biodegradable (AABB) ester polymers (four neutral and one cationic), four organic solvents miscible with water, and eight surfactants were tested for the fabrication of the goal NPs. Depending on the nature of the AABB polymers, organic solvents and surfactants, as well as on the fabrication conditions, the size (Mean Particle Diameter) of the NPs could be tuned within 42 ÷ 398 nm, the zeta-potential within 12.5 ÷ +28 mV. The stability (resuspendability) of the NPs upon storage (at room temperature and refrigerated) was tested as well. In Vitro biocompatibility study of the NPs was performed with four different stable cell lines: A549, HeLa (human); RAW264.7, Hepa 1-6 (murine). Comparing the NPs parameters, their stability upon storage, and the data of biological examinations the best were found: As the AABB polymer, a poly(ester amide) composed of l-leucine, 1,6-hexanediol and sebacic acid–8L6, as a solvent (organic phase—DMSO), and as a surfactant, Tween 20.

Scaffold-based systems have become essential in biomedical research, providing the possibility of building in vitro models that can better mimic tissue/organic physiology. A relatively new family of biomimetics—pseudo-proteins (PPs)—can therefore be considered especially promising in this context. Three different artificial leucine-based LPP films were tested in vitro as potential scaffolding materials. In vitro experiments were performed using two types of cells: primary mouse skin fibroblasts and a murine monocyte/macrophages cell line, RAW264.7. Cell adhesion and cell spreading were evaluated according to morphological parameters via scanning electron microscopy (SEM), and they were assessed according to actin cytoskeleton distribution, which was studied via confocal laser microscopy. Cell proliferation was evaluated via an MTT assay. Cell migration was studied using time-lapse microscopy. SEM images for both types of cells demonstrated prominent adhesion and perfect cell spreading on all three LPPs. Analyses of actin cytoskeleton organization revealed a high number of focal adhesions and prominent motility-associated structures. A certain stimulation of cell proliferation was detected in the cases of all three LPPs, and two of them promoted macrophage migration. Overall, our data suggest that the LPPs used in the study can be considered potential cell-friendly scaffolding materials.

Our research explores leucine-based pseudo-proteins (LPPs) for advanced wound dressings, focusing on their effects on wound healing in an in vitro model. We assessed three types of LPP films for their ability to enhance wound closure rates and modulate cytokine production. They all significantly improved wound closure compared to traditional methods, with the 8L6 and copolymer films showing the most pronounced effects. Notably, the latter exhibited an optimal cytokine profile: an initial burst of pro-inflammatory TNF-α, followed by a controlled release of IL-6 during the proliferative phase and a significant increase in anti-inflammatory IL-10 during remodeling. This balanced cytokine response suggests that the copolymer film not only accelerates wound closure but also supports a well-regulated healing process, potentially reducing fibrosis and abnormal scarring, underscoring the potential of copolymer LPPs as advanced wound dressing materials. Future research will aim to elucidate the specific signaling pathways activated by the copolymer LPP to better understand its mechanism of action. Overall, LPP films offer a promising approach to improving wound care and could lead to more effective treatments for complex wounds.

Natural antioxidants are essential for protecting the body against oxidative stress and exhibit a wide range of biological activities. In this context, forty extracts derived from ten submerged cultivated mushroom species were analyzed for their mycochemical composition, antioxidant capacity, and cytotoxic effects against MCF7 breast cancer cells. Qualitative and quantitative screening revealed that, among the detected classes of bioactive compounds, the extracts were predominantly enriched in flavonoids, terpenoids, and phenolic constituents. Considerable variation was observed in the levels of total phenolics, flavonoids, and ascorbic acid among different species and solvent extracts. The highest total phenolic contents were detected in ethanol and ethyl acetate extracts of G. frondosa (110.0 ± 6.4, 227.6 ± 14.2, and 160.5 ± 5.3 mg GAE/g), while the water extract of F. velutipes also exhibited elevated phenolic levels (119.2 ± 6.5 mg GAE/g). Flavonoid concentrations ranged from 102.5 ± 10.5 to 359.9 ± 2.5 mg QE/g in biomass and culture liquid extracts obtained with organic solvents. Ascorbic acid content was generally highest in ethyl acetate culture liquid extracts, suggesting solvent-dependent enrichment of antioxidant metabolites. Free radical scavenging activity increased in a concentration-dependent manner, reaching inhibition values more than 90% at 20 mg/mL in all tested mushrooms. Cytotoxicity assays demonstrated that extract type, solvent, and incubation time strongly influenced the inhibition of MCF7 cell viability. Ethyl acetate extracts from H. erinaceus, P. ostreatus, T. versicolor, and T. pubescens exhibited the strongest cytotoxic effects, reducing cell viability by up to 70% at higher concentrations. The results demonstrate that mushroom extracts, particularly ethyl acetate extracts, possess significant antioxidant and cytotoxic activities. These findings highlight their potential as promising natural sources of medicinal bioactive compounds for antioxidant and anticancer applications.