Phase equilibrium and crystallization behavior of mixed lipid systems

As complex lipid systems, the phase and crystallization behavior of mixtures of a high-melting milk fat fraction with a low-melting milk fat fraction or canola oil was studied. A turbidity technique was developed to estimate solubility and metastability conditions of these lipid mixtures. Both solubility and metastability of the high-melting milk fat fraction in liquid lipids increased exponentially with temperature. At a given equilibration temperature, liquid phases and solid fractions with nearly identical melting profiles and TAG compositions were obtained regardless of the original concentration of the lipid mixture. The maximum melting temperature (MMT), as measured by DSC, of the liquid phase increased dramatically in the equilibrium temperature range of 27.5–35.0°C but did not change at temperatures below and above this range (down to 25.0°C and up to 40°C in this study). The content of long-chain TAG (C₄₆−C₅₂) increased and short-chain TAG (C₃₆−C₄₀) decreased in the liquid phases as the equilibrium temperature increased. A plot of the TAG group ratio (i.e, long-short-chain TAG) vs. equilibrium temperature was generated to illustrate the phase behavior of the complex lipid system and to represent a solubility curve, from which the supersaturation level for crystallization kinetics was determined. Higher supersaturation and lower temperature resulted in higher nucleation and crystallization rates. Compared to the system with a low-melting milk fat fraction, mixtures of the high-melting milk fat fraction with canola oil had higher nucleation and crystallization rates due to the lower solubility found for this system.     

Oxidative stability of milk drinks containing structured lipids produced from sunflower oil and caprylic acid

Milk drinks containing 5% traditional sunflower oil (SO), randomized lipid (RL) or specific structured lipid (SL) (both produced from SO and tricaprylin/caprylic acid) were compared with respect to their particle size, viscosity and oxidative stability during storage. Furthermore, the effect of adding potential antioxidants EDTA or gallic acid to the milk drink based on SL was investigated. The lipid type significantly affected the oxidative stability of the milk drinks: Milk drink based on SL oxidized faster than milk drink based on RL or SO. The reduced oxidative stability in the SL milk drink could not be ascribed to a single factor, but was most likely influenced by the structure of the lipid and differences in the process applied to produce and purify the lipids. EDTA was a strong antioxidant, while gallic acid did not exert a distinct antioxidative effect in the milk drink based on SL.     

Intracellular Delivery of mRNA for Cell-Selective CRISPR/Cas9 Genome Editing using Lipid Nanoparticles

Messenger RNA (mRNA) is being used as part of an emerging class of biotherapeutics with great promise for preventing and treating a wide range of diseases, as well as encoding programmable nucleases for genome editing. However, mRNA's low stability and immunogenicity, as well as the impermeability of the cell membrane to mRNA greatly limit mRNA's potential for therapeutic use. Lipid nanoparticles (LNPs) are currently one of the most extensively studied nanocarriers for mRNA delivery and have recently been clinically approved for developing mRNA-based vaccines to prevent COVID-19. In this review, we summarize the latest advances in designing ionizable lipids and formulating LNPs for intracellular and tissue-targeted mRNA delivery. Furthermore, we discuss the progress of intracellular mRNA delivery for spatiotemporally controlled CRISPR/Cas9 genome editing by using LNPs. Finally, we provide a perspective on the future of LNP-based mRNA delivery for CRISPR/Cas9 genome editing and the treatment of genetic disorders.     

Ligand Binding Promiscuity of Human Liver Fatty Acid Binding Protein: Structural and Dynamic Insights from an Interaction Study with Glycocholate and Oleate

Human liver fatty acid binding protein (hL‐FABP) has been reported to act as an intracellular shuttle of lipid molecules, thus playing a central role in systemic metabolic homeostasis. The involvement of hL‐FABP in the transport of bile salts has been postulated but scarcely investigated. Here we describe a thorough NMR investigation of glycocholate (GCA) binding to hL‐FABP. The protein molecule bound a single molecule of GCA, in contrast to the 1:2 stoichiometry observed with fatty acids. GCA was found to occupy the large internal cavity of hL‐FABP, without requiring major conformational rearrangement of the protein backbone; rather, this led to increased stability, similar to that estimated for the hL‐FABP:oleate complex. Fast‐timescale dynamics appeared not to be significantly perturbed in the presence of ligands. Slow motions (unlike for other proteins of the family) were retained or enhanced upon binding, consistent with a requirement for structural plasticity for promiscuous recognition.     

酮康唑固体脂质纳米粒体外评价

通过考察酮康唑固体脂质纳米粒体外透皮情况,由结果预测固体脂质纳米粒的发展前景.方法制备酮康唑固体脂质纳米粒,并测定2h、4h、6h、8h、12h、24 h时间点的药物累积透过皮肤与潴留于皮肤的量,通过数据分析评价药物在固体脂质纳米粒里的体外释放及透皮情况.结果药物在固体脂质纳米粒里的皮肤累积透过量低于含药1％的乳膏,但固体脂质纳米粒的皮肤潴留量明显高于含药1％的乳膏.结论固体脂质纳米粒可增加药物在皮肤的局部浓度从而增强药效,同时可减少药物透过真皮进入体循环从而减少副作用,具有良好的发展前景.     

From coffee industry waste materials to skin‐friendly products with improved skin fat levels

Spent coffee grounds (SCG), which are the residue obtained from the treatment of coffee with hot water or steam, can be used for industrial applications, due to the high content in lipids. The cosmetic products might be a suitable application for these types of residues because the barrier properties of the stratum corneum (SC) are largely dependent on the intactness of the lipid lamellae that surrounds the corneocytes. The purpose of this work was to assess the feasibility of using the lipid fraction of SCG extracted with supercritical carbon dioxide in the development of new cosmetic formulations with improved skin lipids (sebum) and hydration. The use of spent coffee lipid extract in cosmetic industry seems to be a suitable approach to recycle the wastes from coffee industry. Emulsion containing 10% of the lipid fraction of SCG (SpentCofOil cream) presented promising characteristics in the improvement of sebum skin levels with a good acceptance by consumers when compared to an emulsion containing 10% w/w of green coffee oil (GreenCofOil cream) and a placebo without coffee oil (NoCofOil cream). Practical applications: In this work, the authors develop and characterize a cream containing 10% of the lipid fraction of SCG extracted with supercritical carbon dioxide with improved skin lipids (sebum) and hydration.     

Extraction of lipids from Yarrowia Lipolytica

BACKGROUND: Microorganisms have often been considered for the production of oils and fats as an alternative to agricultural and animal resources. Extraction experiments were performed using a strain of the yeast Yarrowia lipolytica (Y. lipolytica), a high‐lipid‐content yeast. Three different methods were tested: Soxhlet extraction, accelerated solvent extraction (ASE) and supercritical carbon dioxide (SCCO₂) extraction using ethanol as a co‐solvent. Also, high pressure solubility measurements in the systems ‘CO₂ + yeast oil’ and ‘CO₂ + ethanol + yeast oil’ were carried out. RESULTS: The solubility experiments determined that, at the conditions of the supercritical extractor (40 °C and 20 MPa), a maximum concentration of 10 mg of yeast oil per g of solvent can be expected in pure CO₂. 10% w/w of ethanol in the solvent mixture increased this value to almost 15 mg of yeast oil per g of solvent. Different pretreatments were necessary to obtain satisfactory yields in the extraction experiments. The Soxhlet and the ASE method were not able to complete the lipid extraction. The ‘SCCO₂ + ethanol’ extraction curves revealed the influence of the different pretreatments on the extraction mechanism. CONCLUSION: Evaluating the effectiveness of a given pretreatment, ASE reduced the amount of material and solvent used compared with Soxhlet. In all three cases, the best total extraction performance was obtained for the ethanol‐macerated yeast (EtM). Addition of ethanol to the solvent mixture enhanced the oil solubility. Oil can be extracted from Y. lipolytica in two different steps: a non‐selective ethanol extraction followed by TAG‐selective SCCO₂ purification. © 2012 Society of Chemical Industry     

Challenges in nucleic acid‐lipid films for transfection

The use of surface‐based methods for the delivery of therapeutics has recently generated increasing interest. These platforms have tremendous potential to minimize detrimental side effects associated with systemic delivery by localizing the therapeutic vehicle, and thus provide higher local doses for improved efficacy. Cationic lipids are one of the most commonly used synthetic carriers for the delivery of genetic cargo, such as DNA and RNA. However, reports on the use of lipid‐based films for gene delivery are scarce. Here we investigate the use of a lipid‐based film for the in vitro delivery of plasmid DNA. Solid DNA‐lipid films show very low levels of transfection, while identical complexes prepared for bolus delivery provide high levels of transfection when used directly. We investigate the mechanism, whereby the activity of these solid‐state films is lost and suggest methods for circumventing these challenges and restoring the efficacy of these films as gene delivery platforms. © 2013 American Institute of Chemical Engineers AIChE J, 59: 3203–3213, 2013     

Characteristics of lipid micro- and nanoparticles based on supercritical formation for potential pharmaceutical application

The interest of the pharmaceutical industry in lipid drug delivery systems due to their prolonged release profile, biocompatibility, reduction of side effects, and so on is already known. However, conventional methods of preparation of these structures for their use and production in the pharmaceutical industry are difficult since these methods are usually multi-step and involve high amount of organic solvent. Furthermore, some processes need extreme conditions, which can lead to an increase of heterogeneity of particle size and degradation of the drug. An alternative for drug delivery system production is the utilization of supercritical fluid technique. Lipid particles produced by supercritical fluid have shown different physicochemical properties in comparison to lipid particles produced by classical methods. Such particles have shown more physical stability and narrower size distribution. So, in this paper, a critical overview of supercritical fluid-based processes for the production of lipid micro- and nanoparticles is given and the most important characteristics of each process are highlighted.