Curcumin is a major bioactive component present in turmeric, attributed with a bright yellow color; however, with low solubility and susceptibility to photooxidation, many of the purported health benefits are actually limited. We developed an encapsulation delivery system to produce curcumin nanoparticles using α-cyclodextrin as a carrier and Tween 80 as a surfactant for the purpose of increasing photostability and bioavailability. We used RSM to determine an optimal curcumin nanoparticle (NP) size of 386.4 ± 21.3 nm and entrapment efficiency of 97.4 ± 2.9% for use in functional tests conducted in differentiated Caco-2 cells. Curcumin-NP showed greater (p = 0.001) (74.32 ± 4.20%) stability compared to free curcumin (36.74 ± 3.03%), after treatment with pulsed light, at the highest 20 J/cm2 energy input. The release behavior of curcumin-NPs was pH-dependent and showed a 5 times higher intestinal epithelial cellular uptake and 21.3% decrease of TEER-value, compared to free curcumin (p = 0.017). Compared with free curcumin, the curcumin-NPs had higher antioxidant capacity in all tests while exhibited lower antioxidant capacity loss after pulsed light treatment. Moreover, pre-exposure of curcumin-NPs to pulsed light treatment showed no apparent cytotoxicity effects in Caco-2 cells. In conclusion, using α-cyclodextrin for curcumin-NP encapsulation enhanced both bioavailability and photostability, leading to greater intestinal intracellular antioxidant capacity and no loss in cell viability.

Phytochemical nanoencapsulation is a rapidly emerging technology in the post-harvest processing sector for multiple applications. Edible coatings and aerosol-derived hydrocolloid films are one example of distinct potential uses intended to extend the shelf life of food products by forming a protective barrier that mitigates microbial contamination and moisture loss. Here, we propose a novel approach that combines embedding dual phytochemical-encapsulated nanoparticles within an alpha-cyclodextrin-Tween-based edible coating. An optimized phytochemical mixture of curcumin/piperine, incorporated into nanoparticles (NPs) using a central composite design, resulted in the development of a matrix with alpha-cyclodextrin inclusion and Tween 20 stabilization, which provided minimal particle size (approximate to 285 nm) and maximal encapsulation efficiency (approximate to 98 %). When applied as an aerosol to strawberries and blueberries, the curcumin/piperine NPs significantly extended shelf-life at ambient temperature (22 degrees C) and preserved fruit quality during refrigerated storage (4 degrees C) (P < 0.05). Constructed nanocomplexes exhibited pH-dependent release behavior and varying, but significant (P < 0.05) antimicrobial activity against ,Escherichia coli Bacillus subtilis, Micrococcus luteus, and Enterobacter aerogenes. Texture attributes of fruits, including hardness, cohesiveness, and chewiness were preserved, while enhanced retention of vitamin C corresponded with improved total phenolic content and total antioxidant capacity (P < 0.05). The carry-over antioxidant activity was also observed using differentiated Caco-2 cells in vitro (P < 0.05). Overall, this hydrocolloid-based approach of using alpha-cyclodextrin-Tween nanoencapsulation with curcumin/piperine bioactives enabled a highly effective and food-grade post-harvest strategy for extending the shelf-life and nutritional quality of soft fresh fruits.

COVID-19 infection continues globally with frequent emergence of unfamiliar SARS-CoV-2 variants acting to impair immunity conferred by vaccines. The competitive binding of SARS-CoV-2 spike proteins by angiotensin-converting enzyme 2 (ACE-2) to mimetic and act as a de-coy over that by native ACE-2 receptors on healthy human cells re-mains a practical approach to lessen viral spread. In this study, a therapeutic strategy was developed that targeted gastrointestinal SARS-CoV-2 infection using ACE-2 encapsulated in chi-tosan/tripolyphosphate cross-linked nanoparticles (NPs). Optimization conditions were determined by varying pH (4.0-6.5) and chitosan: ACE-2 mixing ratios (1:1, 1.5:1, 2:1, 2.5:1, 3:1), followed by choice of spray-drying (SD), freeze-drying (FD), or spray-freeze drying (SFD) with varying mannitol concentrations (0, 1:1, and 5:1 of total weight). The optimal formulation was achieved using a pH 5.5 with a mixing chitosan-ACE-2 ratio of 2:1; where ACE-2 loaded NPs had an average particle size of 303.7 nm, polydispersity index (PDI) of 0.21, encapsula-tion efficiency (EE) of 98.4%, zeta potential of 6.8 mV, and ACE-2 loading content (LC) of 28.4%. In general, all drying methods main-tained the spherical shape of the NPs with varying mannitol concen-tration having a significant (P<0.05) effect. After reconstitution, all SD samples had a relatively low yield rate, but the ACE-2 NPs dehydrated specifically by SFD required a lower amount of added mannitol (1:1 of its total weight) and produced a higher yield rate (P<0.05) and similar PDI and EE values, along with relatively good particle size and LC. This formulation also produced a high ACE-2 release and uptake in differentiated Caco-2 cells; thus, representing an effective ACE-2 en-capsulation procedure for use with dry powders.

Blueberry polyphenols exhibit antioxidant and anti-inflammatory properties with a plausible role in preventing chronic diseases. However, applications in food or supplements is limited by low bioavailability. This study was aimed to formulate oleogel-blueberry extract encapsulated systems with chitosan/polyethylene glycol (PEG) nanoparticles (NPs) to enhance resveratrol delivery and bioavailability while retaining bioactivity. NPs were formulated via ionic gelation at pH 5.5 and mixed with soybean (SO), peanut (PO), or hemp oil (HO) to create solid oleogels containing 25 % or 32 % oil with lecithin and sitosterol. Rheological analysis confirmed solid-like properties. HO oleogels showed the highest resveratrol delivery (P < 0.05) and superior chemical stability. Resveratrol release from HO improved bioactive penetration and intracellular antioxidant capacity in cultured Caco-2 intestinal cells, with no toxicity observed. These findings support expanding oleogels as delivery vehicles for blueberry polyphenolics, like resveratrol.

Background/Objectives: COVID-19 infection continues globally, with frequent emergence of unfamiliar SARS-CoV-2 variants acting to impair immunity. The competitive binding of SARS-CoV-2 spike proteins and angiotensin-converting enzyme 2 (ACE-2) can decrease the binding of the virus on native ACE-2 receptors on healthy human cells. It remains a practical approach to lessen viral spread. In this study, a method to encapsulate ACE-2 in the form of chitosan/tripolyphosphate cross-linked nanoparticles (NPs) was developed with emphasis placed on the best dehydration method to secure functional ACE-2 nanoparticles. Methods: Methods: Preparation conditions were assessed by varying pH (4.0–6.5) and the ratio between chitosan and ACE-2 mixing ratios (1:1, 1.5:1, 2:1, 2.5:1, and 3:1). The formulated NPs were then dehydrated using different approaches that included spray-drying (SD), freeze-drying (FD), and spray-freeze drying (SFD) and used varying mannitol concentrations (0, 1:1, and 5:1 of total weight). The mannitol was served as a cryoprotectant in this study. Results: The best formulation achieved used a pH 5.5 with a mixing chitosan–ACE-2 ratio of 2:1, where ACE-2-loaded NPs had an average particle size of 303.7 nm, polydispersity index (PDI) of 0.21, encapsulation efficiency (EE) of 98.4%, and ACE-2 loading content (LC) of 28.4%. After reconstitution, all SD samples had a relatively low yield rate, but the ACE-2 NPs dehydrated specifically using SFD required a lower amount of added mannitol (1:1 of its total weight) and produced a higher yield rate (p < 0.05) and similar PDI and EE values, along with relatively good particle size and LC. This formulation also produced a high ACE-2 release and uptake in differentiated Caco-2 cells, thus representing an effective ACE-2 encapsulation procedure for use with dry powders. Conclusions: This work showed that spray-freeze drying was the best method to dehydrate ACE-2 NPs, using less cryoprotectant to create a significant advantage in terms of greater loading capacity with lower additive requirements.

Red seaweeds are emerging sources of phenolics with potential health benefits. This study presents the first detailed phenolic and antioxidant analysis of Mazzaella japonica using cold plasma-treated water (CPTW) as a green extraction medium. Free, esterified, and insoluble-bound phenolics were extracted following CPTW exposure (10, 20, and 30 min), and individual compounds were identified using HPLC-QTOF-MS/MS. CPTW treatment significantly enhanced phenolic extraction up to 20 min, particularly in the free fraction, while esterified and insoluble-bound fractions declined, suggesting cell wall disruption and hydrolysis promoted phenolic release into the free form. A total of 27 phenolic compounds, mainly phenolic acids, flavonoids, and several phlorotannins, were identified for the first time in M. japonica. Extracts exhibited strong antioxidant activity, especially in the 20-min free fraction, in both chemical assays and Caco-2 cell models. CPTW is a promising, eco-friendly technique to recover functional phenolics from seaweeds for nutraceuticals or other value-added applications.

Angiotensin-converting enzyme 2 (ACE2) is responsible for cell fusion with SARS-CoV viruses. ACE2 is contained in different areas of the human body, including the nasal cavity, which is considered the main entrance for different types of airborne viruses. We took advantage of the roles of ACE2 and the nasal cavity in SARS-CoV-2 replication and transmission to develop a nasal dry powder. Recombinant ACE2 (rhACE2), after a proper encapsulation achieved via spray freeze drying, shows a binding efficiency with spike proteins of SARS-CoV-2 higher than 77 % at quantities lower than 5 mu g/ml. Once delivered to the nose, encapsulated rhACE2 led to viability and permeability of RPMI 2650 cells of at least 90.20 +/- 0.67 % and 47.96 +/- 4.46 %, respectively, for concentrations lower than 1 mg/ml. These results were validated using nasal dry powder containing rhACE2 to prevent or treat infections derived from SARS-CoV-2.

High-pressure processing (HPP) has been employed in the food and pharmaceutical industries for multiple applications, such as microbial inactivation, shelf life extension, homogenizing/stabilizing emulsions, suspensions, gels, and other colloidal systems, cold extraction of meat in crustaceans, the opening of mollusks, etc. However, high pressure is known to affect the stability and the quality of barosensitive (i.e., sensitivity because of the level of pressure) components of the bioproducts, such as proteins. In general, Le Chatelier's principle dictates the fate of high molecular weight polymeric compounds like proteins under high pressure, suggesting a tendency to degrade into simpler monomers. From a structural analysis point of view, this generally results in increased tendencies for the protein to denature from its native state and possibly affect its ability to renature. Protein crystallization is also affected favorably or unfavorably by pressure, depending on the effect of pressure on nucleation and crystal growth steps for the particular type of protein. Protein refolding is another effect whose kinetics can be optimized by pressure. This work discusses the mechanisms of the impact of pressure on protein structure, crystallization refolding, and unfolding, with examples of the application of these processes in recent literature.

Sublingual immunotherapy (SLIT) is a safe and efficacious treatment used to desensitize patients to their food allergies, with recent additional evidence of a durable response persisting several months after discontinuation of therapy.1Schworer SA Kim EH. Sublingual immunotherapy for food allergy and its future directions.Immunotherapy. 2020; 12: 921-931Crossref PubMed Scopus (13) Google Scholar,2Kim EH Keet CA Virkud YV Chin S Ye P Penumarti A et al.Open-label study of the efficacy, safety, and durability of peanut sublingual immunotherapy in peanut-allergic children.J Allergy Clin Immunol. 2023; 151: 1558-1565Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar Studies demonstrating the efficacy of food SLIT have relied almost exclusively on the use of glycerinated food extracts used in skin prick testing. Skin test extracts, however, are costly (eg, currently approximately 100 Canadian dollars/5 mL of a single allergen), can be difficult to access, and are not licensed for SLIT by regulatory agencies such as the US Food and Drug Administration or Health Canada. These factors make food SLIT inaccessible to many families. In contrast, food powders are relatively inexpensive and commercially available. We evaluated the in vitro sublingual protein penetration rates of commercially available food powders in comparison to glycerinated extracts (current standard in clinical SLIT research) for 5 common food allergens: peanut, cashew, egg, cow's milk, and sesame. Glycerinated extracts and powdered forms of the food allergens were purchased for testing. Protein content was quantified using the Bradford protein assay (ThermoFisher Scientific, Rockford, USA; No. A55866). Powders were first dissolved in double distilled water at a concentration of 1 mg/mL. Powder solutions and liquid extracts were diluted as necessary to fit the standard curve. Results were corrected for dilution factors. The contents of the proteins were calculated on the basis of the absorbance at 595 nm (Tecan infinite M200 Pro; Tecan, Männedorf, Switzerland). TR146 cells (MilliporeSigma, Oakville, Canada), a human carcinoma cell line, were used as an in vitro model of sublingual mucosa since they can form a stratified nonkeratinized epithelium and have been used as a suitable in vitro model for human buccal drug delivery permeability studies.3Lin GC Leitgeb T Vladetic A Friedl HP Rhodes N Rossi A et al.Optimization of an oral mucosa in vitro model based on cell line TR146.Tissue Barriers. 2020; 81748459Crossref Scopus (25) Google Scholar,4Sander C Nielsen HM Jacobsen J. Buccal delivery of metformin: TR146 cell culture model evaluating the use of bioadhesive chitosan discs for drug permeability enhancement.Int J Pharm. 2013; 458: 254-261Crossref Scopus (46) Google Scholar Approval for the use of human cell lines was obtained from the University of British Columbia (B17-0247-A003). TR146 cells were cultivated in a cell culture dish (100 mm diameter) using Nutrient Mixture F-12 Ham medium containing 10% fetal calf serum, 100 IU/mL penicillin, and 100 μg/mL streptomycin. The culture was kept at an environment of 37°C, 95% relative humidity with 5% carbon dioxide. The cell culture medium was changed every 2 days. The cells were subcultured every 7 days using 0.05% trypsin solution. TR146 cells were seeded in a 24-well Millicell Hanging Cell Culture Insert (Sigma, Darmstadt, Germany) at a density of 5 × 104 cells/cm2 and placed in the 24-well plates. The medium was changed every 2 days for 30 days before the test. The formation of the monolayers was confirmed before and after the experiment by transepithelial electric resistance using the Millicell-ERS-2 system (Millipore Corporation, Bedford, USA). The experiment was conducted from the apical to the basolateral chamber and Hanks' Balanced Salt Solution with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer (HBSS-HEPES) was used as media. All solid extracts and liquid extracts were diluted by the media with the same protein content and added to the apical chamber. Samples (50 μL) from the basolateral chamber were obtained at 0.5, 1, 2, and 4 hours with 37°C incubation. The permeated protein from the samples was analyzed by Bradford protein assay. The protein content was calculated as the measured protein content minus any protein detected in the blank control samples, which contained no protein from food extracts or powder solutions. All samples were tested in triplicate and averages (±SD) are presented. Two-way repeated analysis of variance tests was applied to determine differences in penetration rates between food powders and glycerinated extracts at 0.5, 1, 2, and 4 hours after inoculation. If statistically significant differences were detected among means, Tukey tests were applied. A 2-sided significance level of 0.05 was applied. Data were analyzed with Stata BE/18.0 (Stata Corp, College Station, Texas). In comparisons of the measured total protein content with the reported total protein content, more variability was observed for glycerinated extracts (between 64% and 236% of reported total protein content) than in the food powders (between 83% and 114% of the reported total protein content) (Table 12Kim EH Keet CA Virkud YV Chin S Ye P Penumarti A et al.Open-label study of the efficacy, safety, and durability of peanut sublingual immunotherapy in peanut-allergic children.J Allergy Clin Immunol. 2023; 151: 1558-1565Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar,5Kim EH Yang L Ye P Guo R Li Q Kulis MD et al.Long-term sublingual immunotherapy for peanut allergy in children: clinical and immunologic evidence of desensitization.J Allergy Clin Immunol. 2019; 144 (1320-1326.e1)Abstract Full Text Full Text PDF Scopus (89) Google Scholar,6Gendo K Orden T Tevrizian A Jacobs J Mozelsio N Gilbert K et al.Food sublingual immunotherapy using consistent, cheaper, and customizable oral immunotherapy solutions.J Asthma Allergy. 2021; 14: 467-470Crossref PubMed Scopus (0) Google Scholar). There was also wide variability in the calculated volume of glycerinated extract required to provide a goal SLIT maintenance dose of 2- to 4-mg dose of protein among the different food extracts (Table 12Kim EH Keet CA Virkud YV Chin S Ye P Penumarti A et al.Open-label study of the efficacy, safety, and durability of peanut sublingual immunotherapy in peanut-allergic children.J Allergy Clin Immunol. 2023; 151: 1558-1565Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar,5Kim EH Yang L Ye P Guo R Li Q Kulis MD et al.Long-term sublingual immunotherapy for peanut allergy in children: clinical and immunologic evidence of desensitization.J Allergy Clin Immunol. 2019; 144 (1320-1326.e1)Abstract Full Text Full Text PDF Scopus (89) Google Scholar,6Gendo K Orden T Tevrizian A Jacobs J Mozelsio N Gilbert K et al.Food sublingual immunotherapy using consistent, cheaper, and customizable oral immunotherapy solutions.J Asthma Allergy. 2021; 14: 467-470Crossref PubMed Scopus (0) Google Scholar).Table 1Total Protein and In Vitro Protein Penetration Rates in Food Powders and Glycerinated Extract for 5 AllergensMeasured and reported total protein contentPreparationMeasured total protein content (mean ± SD)Reported total protein contentaReported total protein content was obtained from food manufacturers nutrition facts food labels, published data, and/or by means of communications from the manufacturer, when available. Reported total protein was not available for cashew and sesame extracts.Percent of reported total protein contentCalculated volume of extract needed to deliver 2 mg proteinbValues calculated as goal sublingual immunotherapy dose of 2 mg protein or 4 mg protein (as per previously published protocols by Kim et al2,5) divided by the mean measured total protein content (mg/mL), as determined by Bradford assay.Calculated volume of extract needed to deliver 4 mg proteinbValues calculated as goal sublingual immunotherapy dose of 2 mg protein or 4 mg protein (as per previously published protocols by Kim et al2,5) divided by the mean measured total protein content (mg/mL), as determined by Bradford assay.ExtractscGlycerinated extracts (1:20 w/v fully dissolved in 0.2% phenol and 50% glycerinated saline) of peanut (Arachis hypogaea), cashew (Anacardium occidentale), egg (Gallus domesticus), cow's milk (Bos domesticus), and sesame (Sesamum indicum) were obtained from Stallergenes Greer Canada (London, Canada). Peanut7.75 ± 0.001 mg/mLapprox 4-8 mg/mLdReported total protein content obtained from Gendo et al.697-1940.26 mL0.52 mL Cashew10.42 ± 0.002 mg/mLN/AN/A0.19 mL0.38 mL Egg4.71 ± 0.03 mg/mL2-4 mg/mLdReported total protein content obtained from Gendo et al.6118-2360.42 mL0.85 mL Cow's milk2.56 ± 0.01 mg/mL2-4 mg/mLdReported total protein content obtained from Gendo et al.664-1280.78 mL1.56 mL Sesame0.88 ± 0.002 mg/mLN/AN/A2.27 mL4.55 mLPowderseThe following brands of powders were purchased commercially: Sahah Naturals PB&Me Original Powdered Peanut Butter (Montreal, Canada), Nuts.com Cashew Flour (Cranford, USA), Hoosier Hill Farm Whole Egg Granules (Fort Wayne, USA), Carnation Instant Skim Milk Powder (Markham, Canada), and Kevala Organic Sesame Flour (Dallas, USA). Peanut0.33 ± 0.0004 mg/mg0.40 mg/mg83 Cashew0.20 ± 0.002 mg/mg0.18 mg/mg111 Egg0.57 ± 0.004 mg/mg0.50 mg/mg114 Cow's milk0.36 ± 0.003 mg/mg0.36 mg/mg100 Sesame0.44 ± 0.2 mg/mg0.40 mg/mg110Protein penetration rate, mean (SD)Time point, hPeanutCow's MilkCashewEggSesameExtractcGlycerinated extracts (1:20 w/v fully dissolved in 0.2% phenol and 50% glycerinated saline) of peanut (Arachis hypogaea), cashew (Anacardium occidentale), egg (Gallus domesticus), cow's milk (Bos domesticus), and sesame (Sesamum indicum) were obtained from Stallergenes Greer Canada (London, Canada).PowdereThe following brands of powders were purchased commercially: Sahah Naturals PB&Me Original Powdered Peanut Butter (Montreal, Canada), Nuts.com Cashew Flour (Cranford, USA), Hoosier Hill Farm Whole Egg Granules (Fort Wayne, USA), Carnation Instant Skim Milk Powder (Markham, Canada), and Kevala Organic Sesame Flour (Dallas, USA).ExtractcGlycerinated extracts (1:20 w/v fully dissolved in 0.2% phenol and 50% glycerinated saline) of peanut (Arachis hypogaea), cashew (Anacardium occidentale), egg (Gallus domesticus), cow's milk (Bos domesticus), and sesame (Sesamum indicum) were obtained from Stallergenes Greer Canada (London, Canada).PowdereThe following brands of powders were purchased commercially: Sahah Naturals PB&Me Original Powdered Peanut Butter (Montreal, Canada), Nuts.com Cashew Flour (Cranford, USA), Hoosier Hill Farm Whole Egg Granules (Fort Wayne, USA), Carnation Instant Skim Milk Powder (Markham, Canada), and Kevala Organic Sesame Flour (Dallas, USA).ExtractcGlycerinated extracts (1:20 w/v fully dissolved in 0.2% phenol and 50% glycerinated saline) of peanut (Arachis hypogaea), cashew (Anacardium occidentale), egg (Gallus domesticus), cow's milk (Bos domesticus), and sesame (Sesamum indicum) were obtained from Stallergenes Greer Canada (London, Canada).PowdereThe following brands of powders were purchased commercially: Sahah Naturals PB&Me Original Powdered Peanut Butter (Montreal, Canada), Nuts.com Cashew Flour (Cranford, USA), Hoosier Hill Farm Whole Egg Granules (Fort Wayne, USA), Carnation Instant Skim Milk Powder (Markham, Canada), and Kevala Organic Sesame Flour (Dallas, USA).ExtractcGlycerinated extracts (1:20 w/v fully dissolved in 0.2% phenol and 50% glycerinated saline) of peanut (Arachis hypogaea), cashew (Anacardium occidentale), egg (Gallus domesticus), cow's milk (Bos domesticus), and sesame (Sesamum indicum) were obtained from Stallergenes Greer Canada (London, Canada).PowdereThe following brands of powders were purchased commercially: Sahah Naturals PB&Me Original Powdered Peanut Butter (Montreal, Canada), Nuts.com Cashew Flour (Cranford, USA), Hoosier Hill Farm Whole Egg Granules (Fort Wayne, USA), Carnation Instant Skim Milk Powder (Markham, Canada), and Kevala Organic Sesame Flour (Dallas, USA).ExtractcGlycerinated extracts (1:20 w/v fully dissolved in 0.2% phenol and 50% glycerinated saline) of peanut (Arachis hypogaea), cashew (Anacardium occidentale), egg (Gallus domesticus), cow's milk (Bos domesticus), and sesame (Sesamum indicum) were obtained from Stallergenes Greer Canada (London, Canada).PowdereThe following brands of powders were purchased commercially: Sahah Naturals PB&Me Original Powdered Peanut Butter (Montreal, Canada), Nuts.com Cashew Flour (Cranford, USA), Hoosier Hill Farm Whole Egg Granules (Fort Wayne, USA), Carnation Instant Skim Milk Powder (Markham, Canada), and Kevala Organic Sesame Flour (Dallas, USA).0.536.7 ± 1.2%30.3 ± 1.5%42.1 ± 0.5%40.7 ± 0.9%28.0 ± 0.7%32.2 ± 0.6%40.4 ± 1.0%40.7 ± 0.9%38.5 ± 2.9%24.2 ± 1.1%182.2 ± 2.0%72.1 ± 1.5%89.5 ± 1.9%84.3 ± 1.5%74.0 ± 2.3%76.3 ± 3.2%86.0 ± 1.0%fValues in each column are statistically different (P < .05).84.3 ± 1.5%fValues in each column are statistically different (P < .05).82.4 ± 1.5%71.4 ± 1.8%289.8 ± 1.5%80.0 ± 2.0%92.8 ± 4.9%92.6 ± 2.5%91.0 ± 1.4%81.2 ± 1.5%92.6 ± 2.5%fValues in each column are statistically different (P < .05).91.7 ± 1.4%fValues in each column are statistically different (P < .05).85.5 ± 1.4%78.3 ± 2.1%495.5 ± 1.5%96.1 ± 1.5%94.9 ± 3.2%93.8 ± 0.7%94.9 ± 1.0%91.1 ± 1.5%96.5 ± 1.7%fValues in each column are statistically different (P < .05).93.8 ± 0.7%fValues in each column are statistically different (P < .05).92.1 ± 1.1%93.8 ± 1.1%Abbreviations: approx, approximately; N/A, not applicable; w/v, weight by volume.a Reported total protein content was obtained from food manufacturers nutrition facts food labels, published data, and/or by means of communications from the manufacturer, when available. Reported total protein was not available for cashew and sesame extracts.b Values calculated as goal sublingual immunotherapy dose of 2 mg protein or 4 mg protein (as per previously published protocols by Kim et al2Kim EH Keet CA Virkud YV Chin S Ye P Penumarti A et al.Open-label study of the efficacy, safety, and durability of peanut sublingual immunotherapy in peanut-allergic children.J Allergy Clin Immunol. 2023; 151: 1558-1565Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar,5Kim EH Yang L Ye P Guo R Li Q Kulis MD et al.Long-term sublingual immunotherapy for peanut allergy in children: clinical and immunologic evidence of desensitization.J Allergy Clin Immunol. 2019; 144 (1320-1326.e1)Abstract Full Text Full Text PDF Scopus (89) Google Scholar) divided by the mean measured total protein content (mg/mL), as determined by Bradford assay.c Glycerinated extracts (1:20 w/v fully dissolved in 0.2% phenol and 50% glycerinated saline) of peanut (Arachis hypogaea), cashew (Anacardium occidentale), egg (Gallus domesticus), cow's milk (Bos domesticus), and sesame (Sesamum indicum) were obtained from Stallergenes Greer Canada (London, Canada).d Reported total protein content obtained from Gendo et al.6Gendo K Orden T Tevrizian A Jacobs J Mozelsio N Gilbert K et al.Food sublingual immunotherapy using consistent, cheaper, and customizable oral immunotherapy solutions.J Asthma Allergy. 2021; 14: 467-470Crossref PubMed Scopus (0) Google Scholare The following brands of powders were purchased commercially: Sahah Naturals PB&Me Original Powdered Peanut Butter (Montreal, Canada), Nuts.com Cashew Flour (Cranford, USA), Hoosier Hill Farm Whole Egg Granules (Fort Wayne, USA), Carnation Instant Skim Milk Powder (Markham, Canada), and Kevala Organic Sesame Flour (Dallas, USA).f Values in each column are statistically different (P < .05). Open table in a new tab Abbreviations: approx, approximately; N/A, not applicable; w/v, weight by volume. Sublingual protein penetration rates were significantly different by time point (0.5, 1, 2, and 4 hours) for all foods tested (P < .05) (Table 12Kim EH Keet CA Virkud YV Chin S Ye P Penumarti A et al.Open-label study of the efficacy, safety, and durability of peanut sublingual immunotherapy in peanut-allergic children.J Allergy Clin Immunol. 2023; 151: 1558-1565Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar,5Kim EH Yang L Ye P Guo R Li Q Kulis MD et al.Long-term sublingual immunotherapy for peanut allergy in children: clinical and immunologic evidence of desensitization.J Allergy Clin Immunol. 2019; 144 (1320-1326.e1)Abstract Full Text Full Text PDF Scopus (89) Google Scholar,6Gendo K Orden T Tevrizian A Jacobs J Mozelsio N Gilbert K et al.Food sublingual immunotherapy using consistent, cheaper, and customizable oral immunotherapy solutions.J Asthma Allergy. 2021; 14: 467-470Crossref PubMed Scopus (0) Google Scholar). No significant differences were observed in penetration rates by food form (extract vs powder) for peanut, cow's milk, cashew, or sesame (P > .05) (Table 12Kim EH Keet CA Virkud YV Chin S Ye P Penumarti A et al.Open-label study of the efficacy, safety, and durability of peanut sublingual immunotherapy in peanut-allergic children.J Allergy Clin Immunol. 2023; 151: 1558-1565Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar,5Kim EH Yang L Ye P Guo R Li Q Kulis MD et al.Long-term sublingual immunotherapy for peanut allergy in children: clinical and immunologic evidence of desensitization.J Allergy Clin Immunol. 2019; 144 (1320-1326.e1)Abstract Full Text Full Text PDF Scopus (89) Google Scholar,6Gendo K Orden T Tevrizian A Jacobs J Mozelsio N Gilbert K et al.Food sublingual immunotherapy using consistent, cheaper, and customizable oral immunotherapy solutions.J Asthma Allergy. 2021; 14: 467-470Crossref PubMed Scopus (0) Google Scholar). For egg, there was a significant difference in penetration rates by food form (P = .0168), with penetration rates of the glycerinated egg extract significantly higher than the powder at 1 hour (86.0 ± 1.0% vs 84.3 ± 1.5%), 2 hours (92.6 ± 2.5% vs 91.7 ± 1.4%), and 4 hours of incubation (96.5 ± 1.7% vs 93.8 ± 0.7%) (P < .05), but not at 0.5 hours (40.4 ± 1.0% vs 40.7 ± 0.9%) (P > .05) (Table 12Kim EH Keet CA Virkud YV Chin S Ye P Penumarti A et al.Open-label study of the efficacy, safety, and durability of peanut sublingual immunotherapy in peanut-allergic children.J Allergy Clin Immunol. 2023; 151: 1558-1565Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar,5Kim EH Yang L Ye P Guo R Li Q Kulis MD et al.Long-term sublingual immunotherapy for peanut allergy in children: clinical and immunologic evidence of desensitization.J Allergy Clin Immunol. 2019; 144 (1320-1326.e1)Abstract Full Text Full Text PDF Scopus (89) Google Scholar,6Gendo K Orden T Tevrizian A Jacobs J Mozelsio N Gilbert K et al.Food sublingual immunotherapy using consistent, cheaper, and customizable oral immunotherapy solutions.J Asthma Allergy. 2021; 14: 467-470Crossref PubMed Scopus (0) Google Scholar). In this study, we observed less variability in the measured protein content of whole food powders than in glycerinated extracts, and statistically significant differences in penetration rates were only observed for egg, with higher rates for glycerinated extract at 1, 2, and 4 hours. However, the percent differences in penetration rates for egg were minimal and would likely have limited practical significance. Since SLIT is believed to promote a tolerogenic environment in the oral cavity through the uptake of allergens by myeloid dendritic cells, several factors, including the influence of pH, saliva, tongue movement, protein weight, allergen solubility, bioavailability, and absorption in the oral cavity1Schworer SA Kim EH. Sublingual immunotherapy for food allergy and its future directions.Immunotherapy. 2020; 12: 921-931Crossref PubMed Scopus (13) Google Scholar,7Guo Y Pratap Singh A Emerging strategies for enhancing buccal and sublingual administration of nutraceuticals and pharamaceuticals.J Drug Deliv Sci Technol. 2019; 52: 440-451Crossref Scopus (38) Google Scholar, 8Yu J Ahmedna M Goktepe I. Peanut protein concentrate: production and functional properties as affected by processing.Food Chem. 2007; 103: 121-129Crossref Scopus (309) Google Scholar, 9Koppelman SJ Smits M Tomassen M De Jong GAH Baumert J Taylor SL et al.Release of major peanut allergens from their matrix under various pH and simulated saliva conditions—ara h2 and ara h6 are readily bio-accessible.Nutrients. 2018; 10: 1281Crossref Scopus (13) Google Scholar, 10Kopper RA Odum NJ Sen M Helm RM Stanley JS Burks AW. Peanut protein allergens: the effect of roasting on solubility and allergenicity.Int Arch Allergy Immunol. 2005; 136: 16-22Crossref PubMed Scopus (0) Google Scholar should be considered when attempting to translate these in vitro findings to in vivo absorption. Furthermore, TR146 cells use endocytosis and passive and active transport mechanisms, similar to in vivo sublingual mucosa, although to varying degrees because of differences in their biologic environments.3Lin GC Leitgeb T Vladetic A Friedl HP Rhodes N Rossi A et al.Optimization of an oral mucosa in vitro model based on cell line TR146.Tissue Barriers. 2020; 81748459Crossref Scopus (25) Google Scholar,4Sander C Nielsen HM Jacobsen J. Buccal delivery of metformin: TR146 cell culture model evaluating the use of bioadhesive chitosan discs for drug permeability enhancement.Int J Pharm. 2013; 458: 254-261Crossref Scopus (46) Google Scholar,7Guo Y Pratap Singh A Emerging strategies for enhancing buccal and sublingual administration of nutraceuticals and pharamaceuticals.J Drug Deliv Sci Technol. 2019; 52: 440-451Crossref Scopus (38) Google Scholar In this study, samples were collected for up to 4 hours, since in vitro absorption by means of TR146 cells is slower than in vivo sublingual absorption (which can occur within minutes because of a high degree of vascularization and a different physiological environment). Further research is needed to understand how the penetration rate in vitro translates to the in vivo sublingual environment since SLIT doses are held under the tongue for only a matter of minutes. Detailed in vitro protein studies for more food allergens, and powder and extract products from a variety of manufacturers are also warranted. Overall, the comparable penetration rates of powders and extracts in vitro support that food SLIT using powders could have clinical usefulness, although further research is needed. Dr Chan has received research support from DBV Technologies; has been a member of advisory boards for Pfizer, Miravo, Medexus, Leo Pharma, Kaleo, DBV, AllerGenis, Sanofi Genzyme, Bausch Health, Avir Pharma, AstraZeneca, and ALK; and was co-lead of the Canadian Society of Allergy and Clinical Immunology oral immunotherapy guidelines. Dr Pratap-Singh holds the BC Ministry of Agriculture Endowed Professorship in Food and Beverage Innovation. Drs Williams, Guo and Soller have no conflicts to disclosure. The authors thank Ally Baaske and Bryant Hartono from the University of British Columbia for their assistance with statistical and laboratory analysis, respectively. This research was funded by the North American Society for Pediatric Gastroenterology, Hepatology, and Nutrition Foundation Council for Pediatric Nutrition Professionals Research Grant; Natural Science and Engineering Research Council of Canada's Discovery Research Grant (No. RGPIN-2018-04735); and Canada Foundation for Innovation John Evans Leaders Fund Award (No. 37498).

Resveratrol is a well described phytochemical with bioactive properties that include antioxidant, antiinflammatory and an affinity to reduce intra-ocular pressure efficiently. The present research describes a method of using ionic gelation using chitosan and polyethylene glycol (PEG) to construct nano-encapsulation of an oil-based blueberry extract, aimed at improving resveratrol bioavailability. Blueberry extract nanoparticles (NPs) were optimally constructed to have a particle size of 343.9 nm and an entrapment efficiency of 98.2% with a polydispersity index (PDI) of 0.231 and a zeta potential of 8.6 mv. Bioavailability accessed using in vitro digestion showed an increase up to 78.6% for resveratrol NP in the blueberry extract. Using differentiated Caco2 cells, the cellular uptake and penetration of NPs displayed highest efficacy compared with free blueberry and blueberry in polyethylene glycol (PEG) (P < 0.05). This study demonstrated the potential of using chitosan/PEG to optimize assembly of NP to improve resveratrol bioavailability when formulated in an oil-based blueberry extract.

Injectable peptides such as insulin, glucagon-like peptide 1 (GLP-1), and their agonists are being increasingly used for the treatment of diabetes. Currently, the most common route of administration is injection, which is linked to patient discomfort as well as being subjected to refrigerated storage and the requirement for efficient supply chain logistics. Buccal and sublingual routes are recognized as valid alternatives due to their high accessibility and easy administration. However, there can be several challenges, such as peptide selection, drug encapsulation, and delivery system design, which are linked to the enhancement of drug efficacy and efficiency. By using hydrophobic polymers that do not dissolve in saliva, and by using neutral or positively charged nanoparticles that show better adhesion to the negative charges generated by the sialic acid in the mucus, researchers have attempted to improve drug efficiency and efficacy in buccal delivery. Furthermore, unidirectional films and tablets seem to show the highest bioavailability as compared to sprays and other buccal delivery vehicles. This advantageous attribute can be attributed to their capability to mitigate the impact of saliva and inadvertent gastrointestinal enzymatic digestion, thereby minimizing drug loss. This is especially pertinent as these formulations ensure a more directed drug delivery trajectory, leading to heightened therapeutic outcomes. This communication describes the current state of the art with respect to the creation of nanoparticles containing peptides such as insulin, glucagon-like peptide 1 (GLP-1), and their agonists, and theorizes the production of mucoadhesive unidirectional release buccal tablets or films. Such an approach is more patient-friendly and can improve the lives of millions of diabetics around the world; in addition, these shelf-stable formulations ena a more environmentally friendly and sustainable supply chain network.

Food fortification can be a solution to anemia in developing countries. A previous study determined that the combination of spray drying, hydroxypropyl-methylcellulose as wall material, and maltodextrin as bulk material, encapsulated iron gluconate achieved the highest bioavailability. However, the addition of vitamin B12 to the hydroxypropyl-methylcellulose/maltodextrin capsules increased the iron cell uptake over the previously reported results. The cell viability, the number of live, healthy cells in a sample, of HepG2, human liver cancer cells, increases by about 17 % for dual-encapsulated iron gluconate and vitamin b12. The cell uptake in Caco2, human colorectal adenocarcinoma cells, is higher by 25 % when using encapsulated iron and vitamin b12 compared to encapsulated iron. The strength of dual-encapsulated iron and vitamin b12 is also confirmed in in-vivo studies. Once fully anemic, young female rats eating food with encapsulated iron gluconate and vitamin b12, show the fastest recovery with respect to rats eating food with encapsulated iron and pure iron. The first needed only five days for their hemoglobulin values to return to normal. The second and the third needed 15 and 21 days, respectively.

Proteins, the building blocks of life, are increasingly being used as therapeutics for treating several diseases. Yet, there are challenges in the delivery of highly labile materials like proteins, which is often circumvented with the help of encapsulation for targeted delivery and enhanced stability. Spray drying technology has recently been employed for encapsulation due to its' low cost and scale-up capabilities, yet the high temperatures of drying air makes the technology unsuitable for proteins. More recently, spray freeze drying has evolved as an emerging technology that combines spray drying with freeze drying by using low temperatures, and is thus suitable for maintaining the stability of proteins. This study investigates the correlation between formulation parameters and the properties of protein encapsulated microparticles prepared by spray freeze drying. Morphology was investigated using microscopic methods, and protein stability was examined using infrared and mass spectrometry. By using bovine serum albumin, we verify that increasing the total weight to 15 mg/ml results in microencapsulates with a projected area equivalent diameter of 100 mu m larger. We demonstrate that some types of amino acids are essential for shell formation; however, glutamine generates an increase in dimer areas in mass spectra of 5.5. D-Mannitol is the suggested carrier for high encapsulation efficiency (above 90 %). The formulation containing polyvinylpyrrolidone, mannitol, and leucine (at 6, 9, and 2 mg/ml, respectively) produced the lowest reduction in the stability of a few types of proteins; deconvoluted infrared peaks show a difference of less than 2% compared to the free protein. Understanding the spray freeze drying phenomenon for protein encapsulation would allow the control over morphological and chemical properties of microparticles containing active proteins.

Nose-to-brain delivery is increasing in popularity as an alternative to other invasive delivery routes. However, targeting the drugs and bypassing the central nervous system are challenging. We aim to develop dry powders composed of nanoparticles-in-microparticles for high efficiency of nose-to-brain delivery. The size of microparticles (between 250 and 350 µm), is desired for reaching the olfactory area, located below the nose-to-brain barrier. Moreover, nanoparticles with a diameter between 150 and 200 nm are desired for traveling through the nose-to-brain barrier. The materials of PLGA or lecithin were used in this study for nanoencapsulation. Both types of capsules showed no toxicology on nasal (RPMI 2650) cells and a similar permeability coefficient (Papp) of Flu-Na, which was about 3.69 ± 0.47 × 10-6 and 3.88 ± 0.43 × 10-6 cm/s for TGF-β-Lecithin and PLGA, respectively. The main difference was related to the location of deposition; the TGF-β-PLGA showed a higher drug deposition in the nasopharynx (49.89 ± 25.90 %), but the TGF-β-Lecithin formulation mostly placed in the nostril (41.71 ± 13.35 %).

Periodontal disease (PD) can be prevented by local or systemic application of epidermal growth factor receptor inhibitors (EGFRIs) that stabilize αvβ6 integrin levels in the periodontal tissue, leading to an increase in the expression of anti-inflammatory cytokines, such as transforming growth factor-β1. Systemic EGFRIs have side effects and, therefore, local treatment of PD applied into the periodontal pockets would be preferrable. Thus, we have developed slow-release three-layered microparticles of gefitinib, a commercially available EGFRI. A combination of different polymers [cellulose acetate butyrate (CAB), Poly (D, L-lactide-co-glycolide) (PLGA) and ethyl cellulose (EC)] and sugars [D-mannose, D-mannitol and D-(+)-trehalose dihydrate] were used for the encapsulation. The optimal formulation was composed of CAB, EC, PLGA, mannose and gefitinib (0.59, 0.24, 0.09, 1, and 0.005 mg/ml, respectively; labeled CEP-gef), and created microparticles of 5.7 ± 2.3 µm in diameter, encapsulation efficiency of 99.98%, and a release rate of more than 300 h. A suspension of this microparticle formulation blocked EGFR phosphorylation and restored αvβ6 integrin levels in oral epithelial cells, while the respective control microparticles showed no effect.

Recent advances in peptide delivery and nanotechnology has resulted in emergence of several non-parenteral administration routes that replace subcutaneous injections associated with patient discomfort. Thiolated biopolymers are relatively new materials being explored to enhance mucoadhesivity and permeability in these efforts, yet their pH dependent reactivity remains an obstacle. This work focussed on improving the mucoadhesivity of thiolated chitosans by activating them with mercaptonicotinic acid, in a bid to create a novel thiomerized chitosan that can open cell tight junctions for application in oral delivery. The synthesized mercaptonicotinic acid activated thiolated chistoan (MNA-TG-chitosan), along with thiolated chitosan (TG-chitosan) and unmodified chitosan were then used to create insulin nanoparticles (insNPs) using spray drying encapsulation process. Use of MNA-TG-chitosan in place of chitosan resulted in reduction of particle size of insNPs from 318 to 277 nm with no significant changes in polydispersity index (~ 0.2), encapsulation efficiency (~ 99%), insulin loading content (~ 25%) and morphology. Results from in-vitro cytotoxicity on TR146, CaCo2 and HepG2 cell lines revealed no significant effects on cell viability at 50–1000 μg/mL concentration. insNPs encapsulated with the new material, MNA-TG-chitosan, resulted in a 1.5-fold and 4.4-fold higher cellular uptake by HepG2 liver cells where insulin is metabolized, approximately 40% and 600% greater insulin transport through TR146 buccal cell monolayers, and 40% and 150% greater apparent permeability than insNPs encapsulated with unmodified chitosan and TG-chitosan respectively. The higher permeation achieved on using MNA-TG chitosan was attributed to the greater opening of the cell tight junction evidenced by reduction of transepithelial electrical resistance of TR146 buccal cell monolayers. This study demonstrates MNA-TG-chitosan as a promising material for improved peptide oral delivery.

Food fortification through iron encapsulated spray-dried microparticles can alleviate the iron deficiency. Here, we analyze the impact of the ratio between the wall and the bulk material, the amount of solids, the type of wall material, and the type of iron compound on the properties of spray dried powder. Optimal formulations contain equal weight percentages of iron compound, bulk material, and wall material. In this case, spray-dried microparticles show an average projected area equivalent diameter of 4.3 mu m with a bioavailability of 60%. Moreover, the encapsulation of iron gluconate reduces the toxicity of Caco-2 and Hep-G2 at any quantities of iron below 12 mg/ml. We verified that most mucoadhesive microparticles include dextran or hydroxyl-methylcellulose; the detachment force between microparticles with a shell made of hydroxyl-methylcellulose and the human intestine is 1.6 N. Moreover, the encapsulation of iron gluconate via spray drying increases Caco-2 iron absorption by 38% compared to non-encapsulated material. An opposite response was observed using iron fumarate, where the absorption was reduced by 31% when the iron compound was encapsulated.

Insulin nanoparticles (NPs) with high loading content have found diverse applications in different dosage forms. This work aimed to evaluate the impact of freeze-drying and spray drying process on the structures of insulin-loaded chitosan nanoparticles, with or without mannitol as cryoprotectants. We also assessed the quality of these nanoparticles by redissolving them. Before dehydration, the chitosan/sodium tripolyphosphate/insulin crosslinked nanoparticles were optimized to 318 nm of particle size, 0.18 of PDI, 99.4% of entrapment efficiency, and 25.01% of loading content. After reconstitution, all nanoparticles, except the one produced by the freeze-drying method without using mannitol, maintained their spherical particle structure. The nanoparticles dehydrated by spray drying without mannitol also showed the smallest mean particle size (376 nm) and highest loading content (25.02%) with similar entrapment efficiency (98.7%) and PDI (0.20) compared to mannitol-containing nanoparticles dehydrated by either spray drying or freeze-drying techniques. The nanoparticles dried by spray drying without mannitol also resulted in the fastest release and highest cellular uptake efficacy of insulin. This work shows that spray drying can dehydrate insulin nanoparticles without the need for cryoprotectants, creating a significant advantage in terms of greater loading capacity with lower additive requirements and operating costs as compared to conventional freeze drying approaches.