Dipalmitoylphosphatidylcholine liposomes were encapsulated with free stigmasterol (ST), stigmasteryl myristate (ME), and stigmasteryl oleate (OE). Their quality was determined using TEM, FT-IR spectroscopy, zeta potential and hydrodynamic diameter. The prepared liposomes were heated at 60 and 180 °C. The degradation of stigmasterol, fatty acids was determined, as was the formation of derivatives. The results show that the liposomes had been prepared successfully. The ST liposomes were the smallest, while the ME and OE liposomes were of similar size. The extent to which the compounds encapsulated in the liposomes degraded depended on their structure. When samples were heated to 60 °C, the degradation of stigmasterol ranged from 11 % in ST to 47 % in OE and 58 % in ME. After heating to 180 °C, the lowest level of degradation of stigmasterol was for OE (51 %), whereas the degradation of stigmasterol in ST and ME was 85 % and 90 %, respectively. In addition, the high level of oxyphytosterols in the samples heated to 180 °C is of concern, especially in the ST and ME samples. The level of SOP in liposomes heated to 60 °C ranged from 1.7 mg/g in liposomes encapsulated with free stigmasterol to 10.4 and 32.9 mg/g in liposomes with stigmasteryl myristate and oleate. After heating to 180 °C, the total content of SOP was much higher, ranging from 68.2 for OE to 111.7 and 135.7 mg/g for ME and ST, respectively.

Background: Phytosterols are bioactive lipids susceptible to oxidation, particularly under thermal stress. Incorporation into liposomes may enhance their stability, while resveratrol—a natural antioxidant—could further limit thermal degradation. Stigmasterol esters, which contain fatty acid residues prone to oxidation, require additional characterization to understand their behavior under heating. Methods: Liposomes composed of dipalmitoylphosphatidylcholine (DPPC) were enriched with free stigmasterol (ST), stigmasteryl myristate (ME), or stigmasteryl oleate (OE), with or without resveratrol (RES). Liposomal systems were characterized using transmission electron microscopy, zeta potential, and hydrodynamic diameter analyses. Samples were heated at 60 °C and 180 °C for 8 h to evaluate stigmasterol degradation, oxyphytosterol (SOP) formation, and decomposition of fatty acid residues in the esters. Results: Liposomes remained structurally stable at 60 °C but underwent marked alterations at 180 °C. ST formed the smallest particles, while ME and OE systems exhibited larger hydrodynamic diameters. Incorporation of resveratrol enhanced thermal and oxidative stability, reducing stigmasterol degradation (7.73–18.86% at 60 °C; 29.66–35.28% at 180 °C) and limiting SOP formation. Differences in the breakdown of myristic versus oleic acid residues highlighted the role of fatty acid type in determining thermal resistance. Conclusions: Resveratrol effectively improves the stability of liposomes containing stigmasterol or its esters and mitigates oxidative damage under thermal stress. Protective effects were particularly evident at moderate temperatures, indicating the potential of resveratrol–phytosterol liposomes as thermally stable delivery systems.

The plasma membrane, composed mostly of lipids and proteins, is a dynamic structure essential for maintaining cellular homeostasis and signaling. Its composition, organization and molecular dynamics have important functional consequences for the cell, while aberrations of its integrity are associated with various human pathologies, including cancers, inflammatory and neurodegenerative diseases. ATP-binding cassette transporter A1 (ABCA1) plays a key role in cellular lipid and cholesterol metabolism, yet its impact on plasma membrane organization and dynamics remains incompletely understood. Using multidisciplinary approaches, we investigated how ABCA1 activity alters plasma membrane dynamics and nanodomain organization in living cells. Fluorescence lifetime imaging microscopy (FLIM) and fluorescence anisotropy showed that active ABCA1 increases membrane homogeneity. Proton nuclear magnetic resonance (¹H NMR) spectroscopy of living cells demonstrated that ABCA1 activity alters membrane fluidity and induces its distinct chemical composition changes. Furthermore, spot variation fluorescence correlation spectroscopy (svFCS) revealed that ABCA1 modulates lipid and protein diffusion dynamics. Our findings confirm and extend previous studies by demonstrating that ABCA1 is one of the key regulators of plasma membrane lipid distribution, influencing its structural and functional organization. By modulating lipid-raft nanodomains, ABCA1 activity may have broader implications for membrane-dependent cellular processes.

Biomembranes, which are the structural and functional basis of the cells of all living organisms, have been an extremely interesting research object for biology and chemistry scientists for years. The multitude of elements constituting the components of natural lipid membranes, however, is associated with interpretation difficulties regarding the nature of the processes taking place in them. A useful research object that is a model of bilamellar biosystems with a significantly simplified composition and at the same time retaining properties that can be a reference point in relation to natural membranes are lipid membranes in the form of one or several component liposomes. It is precisely such systems built of molecules of dipalmitoyl phosphatidylcholine (DPPC) or dipalmitoyl phosphatidylglycerol (DPPG), and analogous systems with the addition of cholesterol (Chol), that were the subject of research in this work. Near-infrared (NIR) vibrational spectroscopy provides a suitable method for the study of the hydrated samples. In most cases it can be alternatively adopted instead of commonly used mid-infrared (MIR) vibrational spectroscopy. This technique was applied for the first time to identify the spectral changes associated with the conformational changes in the hydrophobic region of model lipid bilayers. Trans/gauche isomerization of CH2 groups of lipid hydrocarbon chains is accompanied by characteristic changes in spectral parameters of both νas,s CH2 bands and their first overtones (2νas,s CH2). The heating of all types of analyzed liposomes results in main phase transition (Tm) accompanied by trans to gauche isomerization of CH2 groups of lipid hydrocarbon chains. The NIR-spectroscopy was able to describe in proper way (similar to MIR results) the character of Tm in studied bilayers.