Beclomethasone loaded liposomes enriched with mucin: A suitable approach for the control of skin disorders.

Inflammatory skin disorders are the fourth leading cause of chronic non-fatal conditions, which have a serious impact on the patient quality of life. Due to their treatment with conventional corticosteroids, which often result in poor therapeutic efficacy, relapses and systemic side effects from prolonged therapy, these diseases represent a global burden that negatively impacts the global economy. To avoid these problems and optimize corticosteroid benefits, beclomethasone was loaded into liposome formulations specifically tailored for skin delivery. These formulations were enhanced with mucin (0.1 and 0.5 % w/v) to further ensure prolonged formulation permanence at the site of application. The addition of 0.5 % w/v mucin resulted in the formation of small unilamellar vesicles and multicompartment vesicles. Liposomes and 1mucin-liposomes were smaller (∼48 and ∼61 nm, respectively) and more monodispersed (PI ∼ 0.14 and ∼ 0.17, respectively) than 5mucin-liposomes, which were larger (∼137 nm), slightly polydispersed (PI ∼ 0.23), and less stable during storage (4 months in the dark at 25 °C). Liposomes were negatively charged (∼ -79 mV) irrespective of their composition, and capable of incorporating high amount of beclomethasone (∼ 80 %). In vitro studies on skin fibroblasts and keratinocytes confirmed the high biocompatibility of all formulations (viability ≥ 95 %). However, the use of mucin-liposomes resulted in higher efficacy against nitric oxide production and free radical damage. Finally, topical applications using 12-O-tetradecanoylphorbol-13-acetate-injured skin in vivo experiments showed that only the mucin-enriched formulations could restore healthy conditions within 4 days, underscoring promise as a treatment for skin disorders.

A Cocktail-Based Formula for the Design of Nanosized Cosmeceuticals as Skincare and Anti-Age Products.

Nasco and Bovale grape pomace extracts, alone or in association, were loaded in nanoemulsions tailored for cosmetic application, using Kolliphor®RH40 (kolliphor) as the synthetic surfactant, Olivem®1000 (olivem) as the natural one, and lecithin as the cosurfactant. Pink transparent or milky dispersions, as a function of the used extract and surfactant, were obtained to be used as cosmeceutical serum or milk. The sizes of the nanoemulsion droplets were small (≈77 nm with kolliphor and ≈141 nm with olivem), homogenously dispersed (~0.24 with kolliphor and ~0.16 with olivem), highly negatively charged (≈-43 mV irrespective of the used surfactant) and their stability either on storage or under stressing conditions was affected by the used extract and surfactant. Formulations protected the extracts from the degradation caused by UV exposition, were biocompatible against keratinocytes, protected them against oxidative damages induced using hydrogen peroxide and inhibited the release of nitrite induced in macrophages using the lipopolysaccharide inflammatory stimulus. The overall results underlined the key role played by the composition of the formula to achieve a suitable cosmeceutical for skin care but even for the prevention of premature aging and chronic damages caused by the stressing conditions.

From Grape By-Products to Enriched Yogurt Containing Pomace Extract Loaded in Nanotechnological Nutriosomes Tailored for Promoting Gastro-Intestinal Wellness.

Grape pomace is the main by-product generated during the winemaking process; since it is still rich in bioactive molecules, especially phenolic compounds with high antioxidant power, its transformation in beneficial and health-promoting foods is an innovative challenge to extend the grape life cycle. Hence, in this work, the phytochemicals still contained in the grape pomace were recovered by an enhanced ultrasound assisted extraction. The extract was incorporated in liposomes prepared with soy lecithin and in nutriosomes obtained combining soy lecithin and Nutriose FM06®, which were further enriched with gelatin (gelatin-liposomes and gelatin-nutriosomes) to increase the samples' stability in modulated pH values, as they were designed for yogurt fortification. The vesicles were sized ~100 nm, homogeneously dispersed (polydispersity index < 0.2) and maintained their characteristics when dispersed in fluids at different pH values (6.75, 1.20 and 7.00), simulating salivary, gastric and intestinal environments. The extract loaded vesicles were biocompatible and effectively protected Caco-2 cells against oxidative stress caused by hydrogen peroxide, to a better extent than the free extract in dispersion. The structural integrity of gelatin-nutriosomes, after dilution with milk whey was confirmed, and the addition of vesicles to the yogurt did not modify its appearance. The results pointed out the promising suitability of vesicles loading the phytocomplex obtained from the grape by-product to enrich the yogurt, offering a new and easy strategy for healthy and nutritional food development.

Co-delivering of oleuropein and lentisk oil in phospholipid vesicles as an effective approach to modulate oxidative stress, cytokine secretion and promote skin regeneration.

In this work oleuropein and lentisk oil have been co-loaded in different phospholipid vesicles (i.e., liposomes, transfersomes, hyalurosomes and hyalutransfersomes), to obtain a formulation capable of both inhibiting the production of different markers connected with inflammation and oxidative stress and promoting the skin repair. Liposomes were prepared using a mixture of phospholipids, oleuropein and lentisk oil. Tween 80, sodium hyaluronate or their combination have been added to the mixture to obtain transfersomes, hyalurosomes and hyalutransfersomes. Size, polydispersity index, surface charge and stability on storage was evaluated. The biocompatibility, anti-inflammatory activity and wound healing effect were tested using normal human dermal fibroblasts. Vesicles were small (mean diameter âˆ¼ 130 nm) and homogeneously dispersed (polydispersity index âˆ¼ 0.14), highly negatively charged (zeta potential 02053-64 mV) and capable of loading 20 mg/mL of oleuropein and 75 mg/mL of lentisk oil. The freeze-drying of dispersions with a cryoprotectant permitted to improve their stability on storage. The co-loading of oleuropein and lentisk oil in vesicles inhibited the overproduction of inflammatory markers, especially MMP-1 and IL-6, counteracted the oxidative stress induced in cells using hydrogen peroxide, and promoted the healing of a wounded area performed in vitro in a cell monolayer of fibroblasts. The proposed co-loading of oleuropein and lentisk oil in natural-based phospholipid vesicles may hold promising therapeutic value especially for the treatment of a wide variety of skin disorders.

New Vehiculation Systems of Mometasone Furoate for the Treatment of Inflammatory Skin Diseases.

A pre-formulation study was carried out to obtain liposomal formulations of mometasone furoate as an alternative system to marketed forms of corticosteroid for the treatment of inflammatory skin lesions. Mometasone furoate was loaded in glycerosomes and glyceroethosomes, which were also modified with hyaluronic acid (glyceroethohyalurosomes). Vesicles were designed, elaborated, and characterized, and their biocompatibility, efficacy against oxidative stress and skin lesions were assessed in vitro, in human epidermal cells, and in vivo, in a mouse skin epidermal hyperplasia model. All formulations tested showed great encapsulation efficiency, nanometric size, formed monodispersed systems and a highly negative Z potential. Similar values were obtained over nine months storage at 4 °C, which indicates the great stability of the three types of nanoliposomes at least during the time tested. Among them, 0.1% mometasone furoate glyceroethohyalurosomes were the best formulation to protect cells against oxidative stress and their anti-inflammatory efficacy was confirmed in vivo, being even more effective than the marketed form (Elocom®), as the reduction in the inflammation was even ~15% higher than that achieved with the commercial cream. Selected formulations could be potential candidates as new vehiculation systems for mometasone furoate. The presence of hyaluronic acid in glyceroethohyalurosomes makes them the best candidates in preventing/treating skin inflammatory lesions.

Characterization of Nasco grape pomace-loaded nutriosomes and their neuroprotective effects in the MPTP mouse model of Parkinson's disease.

Grape pomaces have recently received great attention for their richness in polyphenols, compounds known to exert anti-inflammatory and antioxidant effects. These pomaces, however, have low brain bioavailability when administered orally due to their extensive degradation in the gastrointestinal tract. To overcome this problem, Nasco pomace extract was incorporated into a novel nanovesicle system called nutriosomes, composed of phospholipids (S75) and water-soluble maltodextrin (Nutriose® FM06). Nutriosomes were small, homogeneously dispersed, had negative zeta potential, and were biocompatible with intestinal epithelial cells (Caco-2). Nasco pomace extract resulted rich in antioxidant polyphenols (gallic acid, catechin, epicatechin, procyanidin B2, and quercetin). To investigate the neuroprotective effect of Nasco pomace in the subacute 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of Parkinson's disease (PD), Nasco nutriosomes or Nasco suspension was administered intragastrically and their neuroprotective effects were evaluated. Degeneration of nigro-striatal dopaminergic neurons induced by subacute MPTP treatment, the pathological hallmark of PD, was assessed through immunohistochemical evaluation of tyrosine hydroxylase (TH) in the caudate-putamen (CPu) and substantia nigra pars compacta (SNc), and the dopamine transporter (DAT) in CPu. Immunohistochemical analysis revealed that Nasco nutriosomes significantly prevented the reduction in TH- and DAT-positive fibres in CPu, and the number of TH-positive cells in SNc following subacute MPTP treatment, while Nasco suspension counteracted MPTP toxicity exclusively in SNc. Overall, these results highlight the therapeutic effects of Nasco pomace extract when administered in a nutriosome formulation in the subacute MPTP mouse model of PD and validate the effectiveness of the nutriosome preparation over suspension as an innovative nano-drug delivery system for in vivo administration.

Stability and Antioxidant Activity of Hydro-Glyceric Extracts Obtained from Different Grape Seed Varieties Incorporated in Cosmetic Creams.

Grape seeds are agro-industrial by-products, which if improperly managed, may be responsible for socioeconomic and environmental problems. Nevertheless, it is possible to effectively valorize them by means of extraction of the bioactive compounds, especially the antioxidant phenolic molecules, using a safe, green, and environmentally-friendly extractive medium (i.e., hydro-glyceric solution). In the present study, the extraction was performed using seeds from two Lebanese varieties, Obeidi and Asswad Karech, and three international varieties, Marselan, Syrah, and Cabernet Franc. The type and amount of phenolic compounds were identified by High-Performance Liquid Chromatography (HPLC). Marselan was the extract richer in catechins (132.99 ± 9.81 µg/g of dried matter), and it also contained a higher amount of phenolic compounds (49.08 ± 0.03 mg of gallic acid equivalent/g of dry matter and 10.02 ± 0.24 mg of proanthocyanidin content/g of dry matter). The antioxidant capacity of the extracts was assessed using three different colorimetric assays including 2,2-DiPhenyl-1-PicrylHydrazyl (DPPH), CUPRIC ion Reducing Antioxidant Capacity (CUPRAC), and Ferric Reducing Antioxidant Power (FRAP). As expected, Marselan exhibited the highest antioxidant activity; as well, the total phenolic and proanthocyanidin content were the highest. The stability of the Marselan extract incorporated into a commercial cream, was performed at three different temperatures (4, 25, and 50 °C), and four different concentrations (5, 4, 3, 2%), over a period of 4 months, using different methods such as centrifugation, Heat-Shock Cycles, pH, and viscosity. All Marselan hydro-glyceric extract formulations were proven to be stable over the entire 4 months, where the highest stability was achieved at 4 °C and the least at 50 °C. This study supports the suitability of the incorporation of phenolic extracts into commercial creams to enrich the cosmetic industry with effective, natural, and safe skincare products.

A cascade biorefinery for grape marc: Recovery of materials and energy through thermochemical and biochemical processes.

The agro-industrial sector makes a high contribution to greenhouse gas emissions; therefore, proper waste management is crucial to reduce the carbon footprint of the food chain. Hydrothermal carbonization (HTC) is a promising and flexible thermochemical process for converting organic materials into energy and added-value products that can be used in different applications. In this work, grape marc residues before and after an extraction process for recovering polyphenols were hydrothermally treated at 220 °C for 1 h. The resulting hydrochar and process water were investigated to test an innovative cascade approach aimed at a multiple product and energy recovery based on the integration of HTC with anaerobic digestion. The results show that this biorefinery approach applied to grape marc could allow to diversify and integrate its potential valorisation options. The produced hydrochars possess an increased fixed carbon content compared to the feedstock (up to +70 %) and, therefore, can be used in soil, immobilizing carbon in a stable form and partially replacing peat in growing media (up to 5 % in case of hydrochar from grape marc after extraction), saving the consumption of this natural substrate. In addition, energy can be recovered from both hydrochar by combustion and from process water through anaerobic digestion to produce biogas. Hydrochars show good properties as solid fuel similar to lignite, with an energy content of around 27 MJ kg-1 (+30 % compared to the feedstock). The anaerobic digestion of the process water allowed obtaining up to 137 mL of biomethane per gram of fed COD. Finally, while HTC process waters are suitable for biological treatment, attention must be paid to the presence of inhibiting compounds that induce acute toxic effects in aerobic conditions. The proposed approach is consistent with the principles of circular economy and could increase the overall sustainability and resilience of the agro-industrial sector.

Formulation and Testing of Antioxidant and Protective Effect of Hyalurosomes Loading Extract Rich in Rosmarinic Acid Biotechnologically Produced from Lavandula angustifolia Miller.

Culture of plant cells or tissues is a scalable, sustainable, and environmentally friendly approach to obtain extracts and secondary metabolites of uniform quality that can be continuously supplied in controlled conditions, independent of geographical and seasonal variations, environmental factors, and negative biological influences. In addition, tissues and cells can be extracted/obtained from the by-products of other industrial cultivations such as that of Lavandula angustifolia Miller (L. angustifolia), which is largely cultivated for the collection of flowers. Given that, an extract rich in rosmarinic acid was biotechnologically produced starting from cell suspension of L. angustifolia, which was then loaded in hyalurosomes, special phospholipid vesicles enriched with sodium hyaluronate, which in turn are capable of both immobilizing and stabilizing the system. These vesicles have demonstrated to be good candidates for skin delivery as their high viscosity favors their residence at the application site, thus promoting their interaction with the skin components. The main physico-chemical and technological characteristics of vesicles (i.e., mean diameter, polydispersity index, zeta potential and entrapment efficiency of extract in vesicles) were measured along with their biological properties in vitro: biocompatibility against fibroblasts and ability to protect the cells from oxidative stress induced by hydrogen peroxide. Overall, preliminary results disclosed the promising properties of obtained formulations to be used for the treatment of skin diseases associated with oxidative stress and inflammation.

Mouthwash Formulation Co-Delivering Quercetin and Mint Oil in Liposomes Improved with Glycol and Ethanol and Tailored for Protecting and Tackling Oral Cavity.

The aim of this work was the simultaneous loading of quercetin and mint essential oil (mint oil) in phospholipid vesicles specifically tailored to obtain an antibacterial and antioxidant mouthwash. The vesicles were prepared using soy lecithin and Tween 80 as bilayer components, and a mixture of phosphate buffer solution (33%), propylene glycol (33%) and ethanol (33%) as dispersing phase. The formation of regularly shaped, spherical and unilamellar vesicles was confirmed by cryogenic transmission electron microscopy analyses. Similarly, light scattering results disclosed that the size of the vesicles increased by increasing the concentration of mint oil, but at the same time the high amount of mint oil ensured high stability, as the size of these vesicles remained unchanged during 12 months of storage. All tested formulations were highly biocompatible towards epithelial cells and capable of counteracting oxidative cell damages caused by hydrogen peroxide. Moreover, the vesicles prepared with the highest concentration of mint oil inhibited the proliferation of the cariogenic Streptococcus mutans (S. mutans) and Lactobacillus acidophilus (L. acidophilus).

Nasal Spray Formulations Based on Combined Hyalurosomes and Glycerosomes Loading Zingiber officinalis Extract as Green and Natural Strategy for the Treatment of Rhinitis and Rhinosinusitis.

A total green nanotechnological nasal spray has been manufactured and proposed as an alternative treatment of rhinitis and rhinosinusitis. It was obtained by combining the strengthening effect of liposomes on barrier function, the hydrating and lubricating properties of sodium hyaluronan and the anti-inflammatory and antioxidant activities of the extract of Zingiber officinalis. To this purpose, the extract was loaded in special phospholipid vesicles immobilized with hyaluronic acid (hyalurosomes), which were further enriched with glycerol in the water phase. Liposomes and glycerosomes were prepared as well and used as reference. Vesicles were oligolamellar and multicompartment, as confirmed by cryogenic transmission electron microscopy (cryo-TEM) observation, small in size (~140 nm) and negatively charged (~-23 mV). Spray characteristics were evaluated by using the Spraytec® and instant images, from which the plume angle was measured. The range of the droplet size distribution and the narrow spray angle obtained suggest a good nebulization and a possible local deposition in the nasal cavity. In vitro studies performed by using human keratinocytes confirmed the high biocompatibility of vesicles and their ability to effectively counteract oxidative damage on cells induced by hydrogen peroxide. The overall collected data suggest that our vesicles are suitable as nasal spray.

Extraction of the antioxidant phytocomplex from wine-making by-products and sustainable loading in phospholipid vesicles specifically tailored for skin protection.

The present study is aimed at valorizing grape pomace, one of the most abundant winery-making by-products of the Mediterranean area, through the extraction of the main bioactive compounds from the skin of grape pomace and using them to manufacture innovative nanoformulations capable of both avoiding skin damages and promoting skincare. The phytochemicals were recovered through maceration in hydroethanolic solution. Catechin, quercetin, fisetin and gallic acid, which are known for their antioxidant power, were detected as the main compounds of the extract. Liposomes and phospholipid vesicles modified with glycerol or Montanov 82® or a combination of both, were used as carriers for the extract. The vesicles were small (~183 nm), slightly polydispersed (PI ≥ 0.28), and highly negatively charged (~-50 mV). The extract was loaded in high amounts in all vesicles (~100%) irrespective of their composition. The antioxidant activity of the extract, measured by using the DPPH (2,2-Diphenyl-1-picrylhydrazyl) test, was 84 ± 1%, and slightly increased when loaded into the vesicles (~89%, P < 0.05). The grape pomace extract loaded vesicles were highly biocompatible and able to protect fibroblasts (3T3) from the oxidative stress induced by hydrogen peroxide.