Nutraceuticals have gained increasing attention over the last years due to their potential value as therapeutic compounds formulated from natural sources. For instance, there is a wide range of literature about the cardioprotective properties of omega-3 lipids and the antioxidant value of some phenolic compounds, which are related to antitumoral activity. However, the value of nutraceuticals can be limited by their instability under gastric pH and intestinal fluids, their low solubility and absorption. That is why encapsulation is a crucial step in nutraceutical design. In fact, pharmaceutical nanotechnology improves nutraceutical stability and bioavailability through the design and production of efficient nanoparticles (NPs). Lipid nanoparticles protect the bioactive compounds from light and external damage, including the gastric and intestinal conditions, providing a retarded delivery in the target area and guaranteeing the expected therapeutic effect of the nutraceutical. This review will focus on the key aspects of the encapsulation of bioactive compounds into lipid nanoparticles, exploring the pharmaceutical production methods available for the synthesis of NPs containing nutraceuticals. Moreover, the most common nutraceuticals will be discussed, considering the bioactive compounds, their natural source and the described biological properties.
Microalgal biomass is a sustainable source of bioactive lipids with omega-3 fatty acids. The efficient extraction of neutral and polar lipids from microalgae requires alternative extraction methods, frequently combined with biomass pretreatment. In this work, a combined ultrasound and enzymatic process using commercial enzymes Viscozyme, Celluclast, and Alcalase was optimized as a pretreatment method for Nannochloropsis gaditana, where the Folch method was used for lipid extraction. Significant differences were observed among the used enzymatic pretreatments, combined with ultrasound bath or probe-type sonication. To further optimize this method, ranges of temperatures (35, 45, and 55 °C) and pH (4, 5, and 8) were tested, and enzymes were combined at the best conditions. Subsequently, simultaneous use of three hydrolytic enzymes rendered oil yields of nearly 29%, showing a synergic effect. To compare enzymatic pretreatments, neutral and polar lipids distribution of Nannochloropsis was determined by HPLC–ELSD. The highest polar lipids content was achieved employing ultrasound-assisted enzymatic pretreatment (55 °C and 6 h), whereas the highest glycolipid (44.54%) and PE (2.91%) contents were achieved using Viscozyme versus other enzymes. The method was applied to other microalgae showing the potential of the optimized process as a practical alternative to produce valuable lipids for nutraceutical applications.
Damage to the retinal pigment epithelium, Bruch’s membrane and/or tissues underlying macula is known to increase the risk of age-related macular degeneration (AMD). AMD is commonly categorized in two distinct types, namely, the nonexudative (dry form) and the exudative (wet form). Currently, there is no ideal treatment available for AMD. Recommended standard treatments are based on the use of vascular endothelial growth factor (VEGF), with the disadvantage of requiring repeated intravitreal injections which hinder patient’s compliance to the therapy. In recent years, several synthetic and natural active compounds have been proposed as innovative therapeutic strategies against this disease. There is a growing interest in the development of formulations based on nanotechnology because of its important role in the management of posterior eye segment disorders, without the use of intravitreal injections, and furthermore, with the potential to prolong drug release and thus reduce adverse effects. In the same way, 3D bioprinting constitutes an alternative to regeneration therapies for the human retina to restore its functions. The application of 3D bioprinting may change the current and future perspectives of the treatment of patients with AMD, especially those who do not respond to conventional treatment. To monitor the progress of AMD treatment and disease, retinal images are used. In this work, we revised the recent challenges encountered in the treatment of different forms of AMD, innovative nanoformulations, 3D bioprinting, and techniques to monitor the progress.
To develop greener extraction alternatives for microalgae biomass, ultrasound assisted extraction (UAE) and pressurized liquid extraction (PLE) with different biobased solvents were investigated, demonstrating that both techniques are useful alternatives for algal lipid extraction. Specifically, Nannochloropsis gaditana lipids were extracted by UAE and PLE at different temperatures and extraction times with sustainable solvents like 2-Methyltetrahydrofuran (2-MeTHF) and its mixtures with ethanol and other alcohols. The best oil yields for both PLE and UAE of N. gaditana were achieved with the mixture of 2-MeTHF:ethanol (1:3), reaching yields of up to 16.3%, for UAE at 50 °C and up to 46.1% for PLE at 120 °C. Lipid composition of the extracts was analyzed by HPLC-ELSD and by GC-MS to determine lipid species and fatty acid profile, respectively. Different fractionation of lipid species was achieved with PLE and solvent mixtures of different polarity. Thus, for the extraction of glycolipids, ethanolic extracts contained higher amounts of glycolipids and EPA, probably due to the higher polarity of the solvent. The optimized method was applied to microalgae Isochrysis galbana and Tetraselmis chuii showing the potential of mixtures of biobased solvents like 2-methyl-THF and ethanol in different proportions to efficiently extract and fractionate lipids from microalgal biomass.
Carrier-free immobilization is a key process to develop efficient biocatalysts able to catalyze the cell wall degradation in microalgae where the traditional solid supports cannot penetrate. Thus, the insolubilization of commercial Celluclast®, Alcalase®, and Viscozyme® enzymes by carrier-free immobilization and their application in microalgae pretreatment was investigated. In this study, different precipitants at different ratios (ethanol, acetone, and polyethylene glycol 4000) were tested in the first part of the method, to establish the precipitation conditions. The screening of the best precipitant is needed as it depends on the nature of the enzyme. The best results were studied in terms of immobilization yield, thermal stability, and residual activity and were analyzed using scanning electron microscopy. Moreover, a novel strategy was intended including the three enzymes (combi-CLEAs) to catalyze the enzymatic degradation of Nannochloropsis gaditana microalgal cell wall in one pot. The carrier-free immobilized derivatives were 10 times more stable compared to soluble enzymes under the same. At the best conditions showed its usefulness in the pretreatment of microalgae combined with ultrasounds, facilitating the cell disruption and lipid recovery. The results obtained suggested the powerful application of these robust biocatalysts with great catalytic properties on novel and sustainable biomass such as microalgae to achieve cost-effective and green process to extract valuable bioactive compounds.
Microalgae are described as a new source of a wide range of bioactive compounds with health-promoting properties, such as omega-3 lipids. This biomass product is gaining attention mainly due to its potential to accumulate different compounds depending on the species and environment, and it has been commonly recognized as a valuable nutraceutical alternative to fish and krill oils. In this work, we obtained the extract of the microalga Nannochloropsis gaditana, selected on the basis of its content of eicosapentaenoic acid (EPA) and glycolipids, which were determined using GC-MS and high-performance liquid chromatography (HPLC), respectively. To develop an oral formulation for the delivery of the extract, we used a 23 factorial design approach to obtain an optimal lipid nanoparticle formulation. The surfactant and solid lipid content were set as the independent variables, while the particle size, polydispersity index, and zeta potential were taken as the dependent variables of the design. To ensure the potential use of the optimum LN formulation to protect and modify the release of the loaded microalga extract, rheological and differential scanning calorimetry analyses were carried out. The developed formulations were found to be stable over 30 days, with an encapsulation efficiency over 60%.
In recent years, intensive research has been conducted on natural carotenoids extraction using several processes. Conventional extraction methods require high amounts of solvents and a long extraction time. However, pressurized liquid extraction demonstrated to be an interesting method. The extraction efficiencies of pressurized liquid for the recovery of carotenoids, from the green and the orange biomasses of the microalga Dunaliella salina DunaDZ1, are described. Organic solvents were tested including ethanol, n-hexane, ethyl acetate and a mixture of n-hexane:ethanol (3:4). Moreover, three extraction temperatures were used (90, 120 and 150 °C) at constant pressure. Extraction efficiency and extracts characterization were conducted. Results have shown that temperature has a positive effect on extraction yield. HPLC characterization showed that β-carotene is the main carotenoid in the orange biomass, and lutein in the green biomass, with the presence of other minor carotenoids in both biomasses. The highest carotenoid amounts were found in the n-hexane orange biomass extract, with β-carotene isomers as the main carotenoid (138.54 and 357.10 mg/g of dry extract, for cis and trans isomers, respectively). Otherwise, extracts obtained at the lowest tested temperature provided the best carotenoid yields. The best results for the antioxidant activity were obtained at 120 °C for orange biomass ethyl acetate extract.
Microalgae are described as a potential alternative source of bioactive compounds that are environmentally friendly. Dunaliella on its own is well known due to its potential to accumulate large amounts of carotenoids, especially β-carotene. Green and orange biomasses from a new Dunaliella salina strain DunaDZ1 isolated from an Algerian Salt Lake were evaluated for carotenoids composition and antioxidant activity. Ultrasound-Assisted Extraction (UAE) was employed using different extracting solvents. Furthermore, UAE extracts were then analyzed for their chemical composition by TLC, HPLC-DAD and for the antioxidant activity. Lutein was the main carotenoid in the green biomass, with the highest amount for ethyl acetate extract (393.19 mg/g). However, for the orange biomass, the main carotenoid was trans, β-carotene (131.83 mg/g) in the acetonic extract. Moreover, several others carotenoids were detected, belonging to xanthophylls and carotenes. Additionally, ultrasonic-assisted extraction with ethyl acetate produced the extract with the highest antioxidant activity for both D. salina biomasses. These extracts could be used as a natural antioxidant and as an ingredient for functional foods formulation.
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