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The objective of this work was to prepare and characterize liposomes containing co-encapsulated ascorbic acid (AA) and ascorbyl palmitate (AP), as well as to evaluate their stability, cytotoxicity, antioxidant, and antimicrobial activity. Through the pre-formulation studies, it was possible to improve the formulation, as leaving it more stable and with a greater antioxidant activity, resulting in a formulation designated LIP-AAP, with 161 nm vesicle size, 0.215 polydispersity index, -31.7 mV zeta potential, and pH of 3.34. Encapsulation efficiencies were 37% for AA and 79% for AP, and the content was 1 mg/mL for each compound. The optimized liposomes demonstrated stability under refrigeration for 60 days, significant antioxidant activity (31.4 µMol of TE/mL), and non-toxicity, but no antimicrobial effects against bacteria and fungi were observed. These findings confirm that the co-encapsulated liposomes are potent, stable antioxidants that maintain their physical and chemical properties under optimal storage conditions.

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In the food industry context, where fresh cheese stands out as a highly perishable product with a short shelf life, this study aimed to extend its preservation through multi-layer edible coatings. The overall objective was to analyze the biaxial behavior and texture of fresh cheese coated with nanoliposomes encapsulating grape seed tannins (NTs) and polysaccharides (hydroxypropyl methylcellulose; HPMC and kappa carrageenan; KC) using immersion and spray methods, establishing comparisons with uncoated cheeses and commercial samples, including an accelerated shelf-life study. NT, HPMC, and KC were employed as primary components in the multi-layer edible coatings, which were applied through immersion and spray. The results revealed significant improvements, such as a 20% reduction in weight loss and increased stability against oxidation, evidenced by a 30% lower peroxide index than the uncoated samples. These findings underscore the effectiveness of edible coatings in enhancing the quality and extending the shelf life of fresh cheese, highlighting the innovative application of nanoliposomes and polysaccharide blends and the relevance of applying this strategy in the food industry. In conclusion, this study provides a promising perspective for developing dairy products with improved properties, opening opportunities to meet market demands and enhance consumer acceptance.

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Grape seeds are an excellent source of flavonoids and tannins with powerful antioxidant properties. However, the astringency of tannins limits their direct incorporation into food. To overcome this challenge, we investigated the encapsulation of grape seed tannins within nanoliposomes formed by ultrasound cycling. We characterized the nanoliposomes' physicochemical properties, including encapsulation efficiency, antioxidant activity, stability, microstructure, and rheological properties. Our findings reveal that the nanoliposomes exhibited excellent stability under refrigerated conditions for up to 90 days with a mean particle size of 228 ± 26 nm, a polydispersity index of 0.598 ± 0.087, and a zeta potential of -41.6 ± 1.30 mV, maintaining a spherical multilamellar microstructure. Moreover, they displayed high antioxidant activity, with encapsulation efficiencies of 79% for epicatechin and 90% for catechin. This innovative approach demonstrates the potential of using ultrasound-assisted nanoliposome encapsulation to directly incorporate grape seed tannins into food matrices, providing a sustainable and efficient method for enhancing their bioavailability and functionality.

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Edible composite coatings (ECC) formulated from biopolymers that incorporate antioxidant molecules represent an innovative alternative to improve food texture and provide health benefits. Tannins have aroused great interest due to their ability to stabilize suspensions and counteract the effects of free radicals. The mechanical and surface properties are crucial to establishing its quality and applicability. In this study, the objective was to analyze the mechanical and surface properties of ECC made with nanoliposomes that encapsulate grape seed tannins (TLS) and polysaccharides such as hydroxypropylmethylcellulose (HPMC) and kappa carrageenan (KCG) for their future direct application in foods susceptible to oxidation. The inclusion of HPMC or KCG affected the density, showing values in the range of 1010 to 1050 [kg/m3], evidencing significant changes (p < 0.05) in the surface tension in the TLS/FS-HPMC and TLS/FS mixtures. KCG and in the dispersion coefficients, with values in the range of -2.9 to -17.6 [mN/m] in HPS (S1) and -17.6 to -40.9 [mN/m] in PDMS (S2). The TLS/FS-HPMC coating showed higher stiffness and elastic recovery capacity than the TLS/FS-KCG coating, suggesting that the presence of TLS influenced the stiffness of the polymer. HPMC is recommended as a suitable polymer for coating solids, while KCG is more appropriate for suspensions. These findings provide valuable information for directly applying these ECC compounds to food products, potentially offering better preservation and health benefits.

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Nanoliposomes, bilayer vesicles at the nanoscale, are becoming popular because of their safety, patient compliance, high entrapment efficiency, and prompt action. Several notable biological activities of natural essential oils (EOs), including fungal inhibition, are of supreme interest. As developed, multi-compositional nanoliposomes loaded with various concentrations of clove essential oil (CEO) and tea tree oil (TTO) were thoroughly characterized to gain insight into their nano-size distribution. The present work also aimed to reconnoiter the sustainable synthesis conditions to estimate the efficacy of EOs in bulk and EO-loaded nanoliposomes with multi-functional entities. Following a detailed nano-size characterization of in-house fabricated EO-loaded nanoliposomes, the antifungal efficacy was tested by executing the mycelial growth inhibition (MGI) test using Trichophyton rubrum fungi as a test model. The dynamic light scattering (DLS) profile of as-fabricated EO-loaded nanoliposomes revealed the mean size, polydispersity index (PdI), and zeta potential values as 37.12 ± 1.23 nm, 0.377 ± 0.007, and -36.94 ± 0.36 mV, respectively. The sphere-shaped morphology of CEO and TTO-loaded nanoliposomes was confirmed by a scanning electron microscope (SEM). The existence of characteristic functional bands in all tested counterparts was demonstrated by attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. Compared to TTO-loaded nanoliposomes, the CEO-loaded nanoliposomes exhibited a maximum entrapment efficacy of 91.57 ± 2.5%. The CEO-loaded nanoliposome fraction, prepared using 1.5 µL/mL concentration, showed the highest MGI of 98.4 ± 0.87% tested against T. rubrum strains compared to the rest of the formulations.

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Abstract Curcumin is a plant-derived compound with polypharmacological properties that are hampered by its poor solubility, fast degradation, etc. Wound closure complications that follow tooth extraction are numerous, and relatively frequently additional treatment is needed to prevent unwanted process chronification. The present study aims to compare the effects of free and the nanoliposome-encapsulated curcumin on tooth extraction wound closure. The experiments were performed on Wistar rats where both forms of curcumin were applied topically on a tooth extraction wound for seven days. Changes in tissue oxidative stress (malondialdehyde and oxidized proteins concentrations, and catalase activity) and inflammation (nitric oxide levels and myeloperoxidase activity) related parameters were studied three and seven days following the tooth extraction. Also, the extent of pathohistological changes and osteopontin immunohistochemical expression were studied. The obtained results indicate that both forms of curcumin prevent an increase in oxidative stress and inflammation-related parameters in the studied samples at 3-and 7-day time points. Additionally, we found that curcumin diminished tissue inflammatory response and osteopontin expression, while at the same time it caused faster granulation tissue maturation. The encapsulation of curcumin in nanoliposomes proved to be better in improving the extraction wound healing process than the free curcumin, giving this formulation a potential in the pharmaceutical industry.

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Abstract The objective of this study was to evaluate the influence of vitamin C (VC) on the stability of stored liposomes under different climatic conditions. Liposomal formulations containing 1 mg/mL of VC (LIP-VC) and blank formulations (LIP-B) were prepared by the reverse-phase evaporation method. After preparation, they were characterized according to their refractive index, average vesicle diameter, polydispersity index (PDI), zeta potential, pH, content, encapsulation efficiency (EE%), morphology, stability and antioxidant activity. For stability, LIP-VC and LIP-B were stored in different climatic conditions (4 °C, 25 °C and 40 °C) for 30 days. The LIP-VC presented 1.3365 refractive index, 161 nm of mean diameter, 0.231 PDI, -7.3 mV zeta potential, 3.2 pH, 19.4% EE%, spherical morphology, 1 mg/mL of VC content, and antioxidant activity of 12 and 11.4 μmol of TE/mL for the radical DPPH and ABTS+, respectively. During stability, the LIP-B stored in 40 °C showed an instability in the parameters: PDI, vesicle size and zeta potential after 15 days, while the LIP-VC remained stable in its size and PDI for 30 days. After that, it is shown that VC can be used as an antioxidant and stabilizer in liposomes to increase the stability and shelf-life of vesicles.

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The development of greener nano-constructs with noteworthy biological activity is of supreme interest, as a robust choice to minimize the extensive use of synthetic drugs. Essential oils (EOs) and their constituents offer medicinal potentialities because of their extensive biological activity, including the inhibition of fungi species. However, their application as natural antifungal agents are limited due to their volatility, low stability, and restricted administration routes. Nanotechnology is receiving particular attention to overcome the drawbacks of EOs such as volatility, degradation, and high sensitivity to environmental/external factors. For the aforementioned reasons, nanoencapsulation of bioactive compounds, for instance, EOs, facilitates protection and controlled-release attributes. Nanoliposomes are bilayer vesicles, at nanoscale, composed of phospholipids, and can encapsulate hydrophilic and hydrophobic compounds. Considering the above critiques, herein, we report the in-house fabrication and nano-size characterization of bioactive oregano essential oil (Origanum vulgare L.) (OEO) molecules loaded with small unilamellar vesicles (SUV) nanoliposomes. The study was focused on three main points: (1) multi-compositional fabrication nanoliposomes using a thin film hydration-sonication method; (2) nano-size characterization using various analytical and imaging techniques; and (3) antifungal efficacy of as-developed OEO nanoliposomes against Trichophyton rubrum (T. rubrum) by performing the mycelial growth inhibition test (MGI). The mean size of the nanoliposomes was around 77.46 ± 0.66 nm and 110.4 ± 0.98 nm, polydispersity index (PdI) of 0.413 ± 0.015, zeta potential values up to -36.94 ± 0.36 mV were obtained by dynamic light scattering (DLS). and spherical morphology was confirmed by scanning electron microscopy (SEM). The presence of OEO into nanoliposomes was displayed by attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy. Entrapment efficiency values of 79.55 ± 6.9% were achieved for OEO nanoliposomes. In vitro antifungal activity of nanoliposomes tested against T. rubrum strains revealed that OEO nanoliposomes exhibited the highest MGI, 81.66 ± 0.86%, at a concentration of 1.5 µL/mL compared to the rest of the formulations. In summary, this work showed that bioactive OEO molecules with loaded nanoliposomes could be used as natural antifungal agents for therapeutical purposes against T. rubrum.

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Capsaicinoids confer the pungency in chili peppers and are proven to contain many beneficial traits, among them analgesic and anti-inflammatory properties. Capsaicinoids produce a burning sensation when administered and have other secondary effects given their pungency. This is why many drug delivery vehicles have been tested to prove their efficacy in reducing the secondary effects of these compounds while still having its primary beneficial effects. There are many articles that talk about the formulation of drug delivery systems and their properties yet there is little information on the therapeutic effects of applying these drug delivery systems. This review is focused on studies' results that prove efficacy of capsaicinoids specially in those that use them in nano-sized drug delivery systems. Nano-sized carriers have been proven to reduce the secondary effects of capsaicinoids while improving the bioavailability, absorption, anti-inflammatory and analgesic properties of these compounds.

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This work focused on comparing the ability of lecithins with two purity grades regarding their performance in the development of nanoliposomes, as well as their ability to contain and release polar (trans-aconitic acid) and non-polar (quercetin) antioxidant compounds. First, the chemical characterization of both lecithins was carried out through infrared spectroscopy (FTIR), electrospray ionization mass spectrometry (ESI/MS), and modulated differential scanning calorimetry (mDSC). Second, nanoliposomes were prepared by the ethanol injection method and characterized by means of particle size, polydispersity, and zeta potential measurements. Third, the encapsulation efficiency and in vitro release profiles of antioxidants were evaluated. Finally, the antioxidant effect of quercetin and trans aconitic acid in the presence and absence of nanoliposomes was assessed through the oxygen radical absorbance capacity (ORAC) assay. The results showed that, although there are differences in the chemical composition between the two lecithins, these allow the development of nanoliposomes with very similar physicochemical features. Likewise, nanoliposomes elaborated with low purity grade lecithins favored the encapsulation and release of trans-aconitic acid (TAA), while the nanoliposomes made with high purity lecithins favored the encapsulation of quercetin (QCT) and modified its release. Regarding the antioxidant effect, the vehiculization of TAA and QCT in nanoliposomes led to an increase in the antioxidant capability, where QCT showed a sustained effect over time and TAA exhibited a rapidly decaying effect. Likewise, liposomal systems were also found to have a slight antioxidant effect.

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The necessity to develop more efficient, biocompatible, patient compliance, and safer treatments in biomedical settings is receiving special attention using nanotechnology as a potential platform to design new drug delivery systems (DDS). Despite the broad range of nanocarrier systems in drug delivery, lack of biocompatibility, poor penetration, low entrapment efficiency, and toxicity are significant challenges that remain to address. Such practices are even more demanding when bioactive agents are intended to be loaded on a nanocarrier system, especially for topical treatment purposes. For the aforesaid reasons, the search for more efficient nano-vesicular systems, such as nanoliposomes, with a high biocompatibility index and controlled releases has increased considerably in the past few decades. Owing to the stratum corneum layer barrier of the skin, the in-practice conventional/conformist drug delivery methods are inefficient, and the effect of the administered therapeutic cues is limited. The current advancement at the nanoscale has transformed the drug delivery sector. Nanoliposomes, as robust nanocarriers, are becoming popular for biomedical applications because of safety, patient compliance, and quick action. Herein, we reviewed state-of-the-art nanoliposomes as a smart and sophisticated drug delivery approach. Following a brief introduction, the drug delivery mechanism of nanoliposomes is discussed with suitable examples for the treatment of numerous diseases with a brief emphasis on fungal infections. The latter half of the work is focused on the applied perspective and clinical translation of nanoliposomes. Furthermore, a detailed overview of clinical applications and future perspectives has been included in this review.

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Colistin is a re-emergent antibiotic peptide used as a last resort in clinical practice to overcome multi-drug resistant (MDR) Gram-negative bacterial infections. Unfortunately, the dissemination of colistin-resistant strains has increased in recent years and is considered a public health problem worldwide. Strategies to reduce resistance to antibiotics such as nanotechnology have been applied successfully. In this work, colistin was characterized physicochemically by surface tension measurements. Subsequently, nanoliposomes coated with highly deacetylated chitosan were prepared with and without colistin. The nanoliposomes were characterized using dynamic light scattering and zeta potential measurements. Both physicochemical parameters fluctuated relatively to the addition of colistin and/or polymer. The antimicrobial activity of formulations increased by four-fold against clinical isolates of susceptible Pseudomona aeruginosa but did not have antimicrobial activity against multidrug-resistant (MDR) bacteria. Interestingly, the free coated nanoliposomes exhibited the same antibacterial activity in both sensitive and MDR strains. Finally, the interaction of colistin with phospholipids was characterized using molecular dynamics (MD) simulations and determined that colistin is weakly associated with micelles constituted by zwitterionic phospholipids.

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Hybrid encapsulation structures based on ß-carotene-loaded nanoliposomes incorporated within the polymeric ultrathin fibers produced through electrospinning were developed to improve the photostability of the antioxidant. These novel materials were intended to incorporate ß-carotene into water-based food formulations, overcoming the existing limitations associated with its hydrophobic character. Initially, both empty and antioxidant-loaded nanoliposomes were developed and incorporated into polyvinyl alcohol (PVOH) and polyethylene oxide (PEO) solutions. The changes in the solution properties were evaluated to determine their effects on the electrospinning processing. The mixed polymer solutions were subsequently electrospun to produce hybrid nanoliposome-loaded ultrathin fibers. FTIR analysis confirmed the presence of phospholipid molecules inside the electrospun fibers. These ultrathin fibers were evaluated regarding their morphology, diameter, internal ß-carotene distribution and stability against UV irradiation. Liposomal release studies from the electrospun fibers were also undertaken, confirming the presence of the liposomal structures after dissolving the electrospun fibers in water.

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Liposomes are known to be a potent adjuvant for a wide range of antigens, as well as appropriate antigen carriers for antibody generation response in vivo. In addition, liposomes are effective vehicles for peptides and proteins, thus enhancing their immunogenicity. Considering these properties of liposomes and the antigenicity of the Leishmania membrane proteins, we evaluated if liposomes carrying glycosylphosphatidylinositol (GPI)-anchored proteins of Leishmania amazonensis promastigotes could induce protective immunity in BALB/c mice. To assay protective immunity, BALB/c mice were intraperitoneally injected with liposomes, GPI-protein extract (EPSGPI) as well as with the proteoliposomes carrying GPI-proteins. Mice inoculated with EPSGPI and total protein present in constitutive proteoliposomes displayed a post-infection protection of about 70% and 90%, respectively. The liposomes are able to work as adjuvant in the EPSGPI protection. These systems seem to be a promising vaccine prototype for immunisation against leishmaniasis.

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