Bioavailability by design - Vitamin D3 liposomal delivery vehicles.

Vitamin D3 deficiency has serious health consequences, as demonstrated by its effect on severity and recovery after COVID-19 infection. Because of high hydrophobicity, its absorption and subsequent redistribution throughout the body are inherently dependent on the accompanying lipids and/or proteins. The effective oral vitamin D3 formulation should ensure penetration of the mucus layer followed by internalization by competent cells. Isothermal titration calorimetry and computer simulations show that vitamin D3 molecules cannot leave the hydrophobic environment, indicating that their absorption is predominantly driven by the digestion of the delivery vehicle. In the clinical experiment, liposomal vitamin D3 was compared to the oily formulation. The results obtained show that liposomal vitamin D3 causes a rapid increase in the plasma concentration of calcidiol. No such effect was observed when the oily formulation was used. The effect was especially pronounced for people with severe vitamin D3 deficiency.

The Membrane Electrical Potential and Intracellular pH as Factors Influencing Intracellular Ascorbate Concentration and Their Role in Cancer Treatment.

Ascorbate is an important element of a variety of cellular processes including the control of reactive oxygen species levels. Since reactive oxygen species are implicated as a key factor in tumorigenesis and antitumor therapy, the injection of a large amount of ascorbate is considered beneficial in cancer therapy. Recent studies have shown that ascorbate can cross the plasma membrane through passive diffusion. In contrast to absorption by active transport, which is facilitated by transport proteins (SVCT1 and SVCT2). The passive diffusion of a weak acid across membranes depends on the electrostatic potential and the pH gradients. This has been used to construct a new theoretical model capable of providing steady-state ascorbate concentration in the intracellular space and evaluating the time needed to reach it. The main conclusion of the analysis is that the steady-state intracellular ascorbate concentration weakly depends on its serum concentration but requires days of exposure to saturate. Based on these findings, it can be hypothesized that extended oral ascorbate delivery is possibly more effective than a short intravenous infusion of high ascorbate quantities.

The Elucidation of the Molecular Mechanism of the Extrusion Process.

Extrusion is a popular method for producing homogenous population of unilamellar liposomes. The technique relies on forcing a lipid suspension through cylindrical pores in a polycarbonate membrane. The quantification of the extrusion and/or recalibration processes make possible the acquisition of experimental data, which can be correlated with the mechanical properties of the lipid bilayer. In this work, the force needed for the extrusion process was correlated with the mechanical properties of a lipid bilayer derived from other experiments. Measurements were performed using a home-made dedicated device capable of maintaining a stable volumetric flux of a liposome suspension through well-defined pores and to continuously measure the extrusion force. Based on the obtained results, the correlation between the lipid bilayer bending rigidity and extrusion force was derived. Specifically, it was found that the bending rigidity of liposomes formed from well-defined lipid mixtures agrees with data obtained by others using flicker-noise spectroscopy or micromanipulation. The other issue addressed in the presented studies was the identification of molecular mechanisms leading to the formation of unilamellar vesicles in the extrusion process. Finally, it was demonstrated that during the extrusion, lipids are not exchanged between vesicles, i.e., vesicles can divide but no membrane fusion or lipid exchange between bilayers was detected.

Evaluation of the In Vitro Stability of Stimuli-Sensitive Fatty Acid-Based Microparticles for the Treatment of Lung Cancer.

The fatty acid-based microparticles containing iron oxide nanoparticles and paclitaxel (PAX) are a viable proposition for the treatment of lung cancer. The microparticles inhaled as a dry powder can be guided to selected locations using an external magnetic field, and when accumulated there, the active compound release can be triggered by local hyperthermia. However, this general strategy requires that the active compound is released from microparticles and can reach the targeted cells before microparticles are removed. Isothermal titration calorimetry was used to demonstrate that the components of microparticles were released and transferred to albumins and lipid bilayers. The morphology of the measured particulates was studied with scanning electron microscopy and dynamic light scattering. To determine the cytotoxicity of microparticles, cell culture studies were done. It has been shown that the transfer efficiency depends predominantly on the fatty acid composition of microparticles, which, together with the active ingredient, accumulate predominantly in membrane structures after being released from microparticles and before entering the cytoplasm. The release process is sufficient; hence, paclitaxel-loaded microparticles effectively suppressed the proliferation of A549 human lung epithelial cells of malignant origin (IC50 values for both lauric acid-based and myristic/palmitic-based microparticles containing paclitaxel were below 0.375 µg/mL), while reference microparticles were noncytotoxic.

Experimental evidence and physiological significance of the ascorbate passive diffusion through the lipid bilayer.

The diverse range of functions performed by ascorbate in many metabolic processes requires its effective redistribution between various aqueous body compartments. It is believed that this hydrophilic molecule needs protein transporters for crossing the biological membrane barriers. Any effective model reflecting the ascorbate distribution within the body requires bi-directional fluxes, but only the ascorbate transporters facilitating its intake by cells have been identified to date. The cellular efflux of this molecule still lacks proper mechanistic explanation, nevertheless data suggesting possible passive ascorbate transport recently appeared. In the paper, we provide experimental evidences that ascorbate associates efficiently with the lipid bilayer interface and slowly crosses its hydrophobic core. The measured logPmembrane/water and membrane permeability coefficient equal to 3 and 10-7 - 10-8 cm/s, respectively. The ascorbate passive diffusion across the lipid bilayer provides the missing element needed for the construction of a consistent physiological model describing the ascorbate local homeostasis. The model was effectively used for the construction of the mechanistic description of the processes, which facilitate the ascorbate homeostasis in the brain.

The effect of lipid phase on liposome stability upon exposure to the mechanical stress.

Mechanical properties of a lipid bilayer are parameters determined mainly for giant unilamellar vesicles (GUVs). It is not clear if values obtained on the GUV model can be directly translated to submicron large unilamellar vesicles (LUVs). This ambiguity is a major obstacle in exploring the effect of lipid bilayer mechanics on membrane associated processes and effectiveness of liposome-based targeted drug delivery systems. In presented work extrusion, which is a common method to prepare LUVs, was used to study liposomes preparation and stability upon exposure to mechanical stress. The effect of parameters of the extrusion process (temperature, membrane pore size, extrusion force and volumetric flux) on the properties of liposome suspension (average liposome size, polydispersity index and lipid recovery ratio) was determined for model liposomes composed of DPPC lipid. The state of the DPPC lipid bilayer depends on temperature, therefore, the effect of lipid bilayer mechanics on the extrusion process can be quantitated without altering membrane composition. The extrusion process was carried out with the automated extruder delivering quantitative data on the extrusion force and volumetric flux. Obtained results have been interpreted in terms of mechanical properties of the lipid bilayer. Determined mechanical properties of the lipid bilayer and its dependence on temperature are in good agreement with the literature results determined for GUVs. This shows that mechanical properties of the lipid bilayer does not depend on the liposome size in the range from 100 nm to hundreds of microns.

New oral liposomal vitamin C formulation: properties and bioavailability.

Vitamin C is the exogenous compound necessary for a variety of metabolic processes; therefore, the efficient delivery is critical for the maintenance of body homeostasis. Vitamin C pharmacokinetics and low quantities in processed foodstuff, necessitates its continuous supplementation. In the paper, we present the new liposomal formulation of vitamin C free of harmful organic solvents. The formulation was quantitatively characterized with respect to its chemically composition and nano-structuring. The vitamin C accessibility to cells from the formulation was evaluated using evidence derived from experiments performed on cell cultures. Finally, the enhanced bioavailability of vitamin C from the formulation was demonstrated in the medical experiment.