Gastrointestinal Simulation Research Articles

Bioactivity evaluation of microbial exopolysaccharide and its use as an encapsulating agent for Lactobacillus acidophilus (LA-3)

Microbial exopolysaccharides present bioactive and technological characteristics of interest to the food and pharmaceutical industries. This study aimed to evaluate the bioactivity of the exopolysaccharide - EPS-LN60 - obtained from L. plantarum DF60Mi and its use as an encapsulating agent for Lactobacillus acidophilus LA-3. The prebiotic score, antioxidant activity, and cytotoxicity in human tumor and non-tumor cells were determined. Solutions of 0% (F0), 0.5% (F1), and 1.0% (F2) of EPS were used as a coating for alginate capsules containing LA-3, which were evaluated for morphology, survival of LA-3 during refrigerated storage and after exposure to in vitro gastrointestinal conditions. EPS-LN60 had a higher prebiotic capacity (1.45 ± 0.17) compared to the fructooligosaccharide (0.24 ± 0.05) and an antioxidant activity of 1862 ± 23.08 μM FeSO4/g. Furthermore, EPS-LN60 inhibited tumor cell growth by 36%±2.78%. The capsules coated with EPS-LN60 were spherical, uniform, and efficient toward LA-3 survival (85%) after 20 days of storage. F0 did not have viable cells after 10 days. EPS-LN60 provided additional protection to the capsule, preventing loss of viability during gastrointestinal simulation conditions, resulting in survival rates greater than 86%, which was not observed in F0 when submitted to the same conditions. Therefore, the potential of EPS-LN60 was demonstrated in terms of its bioactivity and its use as an encapsulating agent for L. acidophilus LA-3, with the possibility of future exploitation as a functional ingredient by the food and pharmaceutical industries.

Fabricating pectin and chitosan double layer coated liposomes to improve physicochemical stability of beta-carotene and alter its gastrointestinal fate

To improve storage stability and gastrointestinal (GI) stability of liposomes, pectin and chitosan double layer coated liposome (P-C-L) was proposed and optimized using electrostatic deposition technique. The physical-chemical properties and GI fate of the carrier were then investigated in comparison to that of chitosan coated liposomes (C-L) and un-coated liposomes (L). The results showed P-C-L was successfully prepared at 0.2 % chitosan and 0.06 % pectin. It was hydrogen bonds between the amino groups in chitosan and liposomal interfacial region, and the interaction between the carboxyl groups in pectin layer and amino groups in chitosan layer maintained the structure of P-C-L after absorption by electrostatic interaction. The double layer coatings could improve chemical stability of the encapsulated β-carotene (βC), as well as the thermal stability of liposomes. What's more, the permeability of liposomal bilayers and βC release mechanism in simulated GI fluids was changed by the polymer coating. P-C-L exhibited better controlled release for βC than C-L and L, and displayer beneficial effect on delivering bioactive agents passing through intensity tract. This may assistant developing more efficient delivery system for bioactive agent.

Intestine-targeted high internal phase Pickering emulsion formulated using silkworm pupa protein via ultrasonic treatment

High internal phase Pickering emulsions (HIPPEs) stabilized by food grade particles have received much attention as deliver vehicles for bioactives in recent years. In this study, ultrasonic treatment was conducted to regulate the size of silkworm pupa protein (SPP) particles, fabricating oil–in–water (O/W) HIPPEs with intestinal releasability. Briefly, the pretreated SPP and SPP-stabilized HIPPEs were characterized, and the targeting release was investigated using in vitro gastrointestinal simulations and sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Results revealed ultrasonic treatment time was the key factor regulating emulsification performance and stability of HIPPEs. Optimized SPP particles were obtained based on their size and zeta potential of 152.67 nm and 26.77 mV, respectively. With ultrasonic treatment, the hydrophobic groups in the secondary structure of SPP were exposed, facilitating the formation of a stable oil–water interface for HIPPEs. Additionally, SPP-stabilized HIPPE showed high stable against the gastric digestion. The SPP with 70 kDa molecular weight, which was the major interfacial proteins of the HIPPE, can be hydrolyzed by intestinal digestive enzymes, enabling the intestine-targeted release of the emulsion. Overall, in the present study, a facile method was developed to stabilize HIPPEs using solo SPP with ultrasonic treatment to protect and deliver hydrophobic bioactive ingredients.

Geochemical fractionation, bioaccessibility and ecological risk of metallic elements in the weathering profiles of typical skarn-type copper tailings from Tongling, China

Nonferrous metal tailings have long posed a significant threat to the surrounding environment and population. Previous studies have primarily focused on heavy metal pollution in the vicinity of sulfide tailings, while little attention was given to metal mobility and bioavailability within skarn-type tailings profile during weathering. Therefore, this study aimed to investigate the fractionation, bioaccessibility, and ecological risk associated with metallic elements (MEs, including Pb, Cd, Cr, Zn, and Cu) in two representative weathering copper-tailings profiles of Tongling mine (China). This was achieved through the use of mineralogical analyses, BCR extractions (F1: exchangeable, F2: reducible, F3: oxidizable, F4: residual fraction), in-vitro gastrointestinal simulation test (PBET) and risk assessment models. The mineral compositions of two weathering profiles were similar, with quartz and calcite being the dominant minerals, along with minor amounts of siderite, hematite and spangolite. The mean concentration in the tailings profile was approximately 0.31 (Cr), 1.8 (Pb), 12 (Zn), 33 (Cd) or 34 (Cu) times of the local background values (LBVs). The mean content of the bottom weakly-weathering layer in profile was about 0.36 (Cr), 0.91 (Pb), 1.91 (Cd), 2.73 (Zn) or 2.68 (Cu) times of the surface oxide layer, indicating a strong weathering-leaching effect. The average proportion of BCR-F1 fraction for Cd (30.94 %) was the highest among the five MEs, possibly due to its association with calcite. The PBET-extracted fractions for Cd, Zn and Cu were significantly positively correlated with the F1, F2 and F3 fractions of BCR, suggesting that these elements have higher bioavailability/bioaccessibility. The assessment results indicated that Cd posed a higher health risk, while the risk of Cu, Zn, and Pb is relatively low and Cr is safe. In conclusion, this study provides valuable insights into the environmental geochemical behavior and potential risks of MEs in skarn-type non-ferrous metal tailings ponds.

Release of an encrypted, highly potent ACE-inhibitory peptide by enzymatic hydrolysis of moth bean (Vigna aconitifolia) protein.

Dietary approaches for the regulation of blood pressure are the need of the hour. Hence, identifying the foods possessing such activity is gaining importance. With this aim, moth bean (Vigna aconitifolia), an underutilized pulse, was explored for the presence of antihypertensive activity in terms of angiotensin converting enzyme (ACE)-inhibition bioactivity. Defatted moth bean protein concentrate was hydrolyzed by using different proteases including Alcalase, papain, and trypsin, to identify the enzyme producing highly potent ACE inhibitory peptides. The hydrolysate showing the highest ACE inhibitory activity was further fractionated using an ultrafiltration membrane (10, 3 and 1 kDa) based on ACE inhibitory activity. The active fraction was further subjected to the ion-exchange chromatography followed by RP-HPLC and LC-MS/MS analysis for the enrichment and identification of ACE inhibitory peptides. Finally, based on the bioinformatic analysis, few peptides were synthesized and evaluated for ACE inhibitory activity, followed by docking study and molecular dynamic simulation of a peptide with the highest ACE inhibitory activity. Out of the three proteases, Alcalase-derived hydrolysate showed the highest (~59%) ACE inhibition activity. Molecular weight-based fractionation revealed that <1 kDa fraction possessed the highest ACE inhibitory activity. Activity guided separation of 1 kDa fraction using ion-exchange chromatography, RP-HPLC and LC-MS/MS showed the presence of about 45 peptides. Based on the bioinformatic analysis, 15 peptides were synthesized and evaluated for ACE inhibitory activity. Among these, a novel octapeptide FPPPKVIQ showed the highest ACE inhibitory activity (93.4%) with an IC50 of 0.24 μM. This peptide retained about 59% activity post gastrointestinal digestion simulation. A Dixon plot as well as docking studies revealed the uncompetitive inhibitory nature of this peptide with a Ki value of 0.81 μM. Molecular dynamic simulation studies till 100 ns ensured the stability of the ACE-peptide complex. Thus, present study identified a novel potent ACE inhibitory peptide from moth bean that can be incorporated in a functional dietary formulation for regulation of hypertension.

Digested Cinnamon (Cinnamomum verum J. Presl) Bark Extract Modulates Claudin-2 Gene Expression and Protein Levels under TNFα/IL-1β Inflammatory Stimulus.

Epigenetic changes, host-gut microbiota interactions, and environmental factors contribute to inflammatory bowel disease (IBD) onset and progression. A healthy lifestyle may help to slow down the chronic or remitting/relapsing intestinal tract inflammation characteristic of IBD. In this scenario, the employment of a nutritional strategy to prevent the onset or supplement disease therapies included functional food consumption. Its formulation consists of the addition of a phytoextract enriched in bioactive molecules. A good candidate as an ingredient is the Cinnamon verum aqueous extract. Indeed, this extract, subjected to a process of gastrointestinal digestion simulation (INFOGEST), exhibits beneficial antioxidant and anti-inflammatory properties in an in vitro model of the inflamed intestinal barrier. Here, we deepen the study of the mechanisms related to the effect of digested cinnamon extract pre-treatment, showing a correlation between transepithelial electrical resistance (TEER) decrement and alterations in claudin-2 expression under Tumor necrosis factor-α/Interleukin-1β (TNF-α/IL-1) β cytokine administration. Our results show that pre-treatment with cinnamon extract prevents TEER loss by claudin-2 protein level regulation, influencing both gene transcription and autophagy-mediated degradation. Hence, cinnamon polyphenols and their metabolites probably work as mediators in gene regulation and receptor/pathway activation, leading to an adaptive response against renewed insults.

Effects of lyoprotectant and encapsulated Lactobacillus acidophilus KBL409 on freeze-drying and storage stability

The purpose of this study was to optimize the encapsulated formulation and freeze-drying conditions of Lactobacillus acidophilus KBL409 to improve shelf-life. The optimum freeze-drying conditions was 8% sucrose as a lyoprotectant at −30 °C. In addition, the free, alginate-encapsulated (Al-), and alginate/chitosan-encapsulated (Al/Chi-) L. acidophilus KBL409 with or without sucrose were prepared and evaluated for the simulated gastrointestinal (SGI) tract, moisture characteristics, and storage stability. The freeze-dried Al/Chi-cells with sucrose displayed the highest survival rate of 0.14% in the SGI tract (p < 0.05). The cells showed sigmoidal sorption isotherms and moisture content of 0.084 kg water/kg solid for Brunauer-Emmett-Teller-monolayer. At 4 °C/60% RH, the glass transition temperature was 12.55 °C, which allowed for 12-week period of glassy state and the highest storage viability of 22.35%. In conclusion, the viability of Lactobacillus acidophilus KBL409 faces distinct challenges for freeze-drying and storage, and encapsulation with lyoprotectant affects various stability and quality attributes of microorganisms.

Metformin and curcumin co-encapsulated chitosan/alginate nanoparticles as effective oral carriers against pain-like behaviors in mice

Nanotechnology plays an integral role in multimodal analgesia. In this study, we co-encapsulated metformin (Met) and curcumin (Cur) into chitosan/alginate (CTS/ALG) nanoparticles (NPs) at their synergistic drug ratio by applying response surface methodology. The optimized Met-Cur-CTS/ALG-NPs were achieved with Pluronic® F-127 2.33 % (w/v), Met 5.91 mg, and CTS:ALG mass ratio 0.05:1. The prepared Met-Cur-CTS/ALG-NPs had 243 nm particle size, −21.6 mV zeta potential, 32.6 and 44.2 % Met and Cur encapsulations, 19.6 and 6.8 % Met and Cur loading, respectively, and 2.9:1 Met:Cur mass ratio. Met-Cur-CTS/ALG-NPs displayed stability under simulated gastrointestinal (GI) fluid conditions and during storage. In vitro release study of Met-Cur-CTS/ALG-NPs in simulated GI fluids showed sustained release, with Met exhibiting Fickian diffusion and Cur demonstrating non-Fickian diffusion following the Korsmeyer-Peppas model. Met-Cur-CTS/ALG-NPs exhibited increased mucoadhesion and improved cellular uptake in Caco-2 cells. Additionally, Met-Cur-CTS/ALG-NPs exhibited better anti-inflammatory effects in lipopolysaccharide-stimulated RAW 264.7 macrophage and BV-2 microglial cells than the equivalent amount of the Met-Cur physical mixture, indicating a greater ability to modulate peripheral and central immune mechanisms of pain. In the mouse formalin-induced pain model, Met-Cur-CTS/ALG-NPs administered orally exhibited better attenuation of pain-like behaviors and proinflammatory cytokine release compared to the Met-Cur physical mixture. Furthermore, Met-Cur-CTS/ALG-NPs did not induce significant side effects in mice at therapeutic doses. Altogether, the present study establishes a CTS/ALG nano-delivery system for Met-Cur combination against pain with improved efficacy and safety.

Capsules with Ileocolonic-Targeted Release of Vitamin B2, B3, and C (ColoVit) Intended for Optimization of Gut Health: Development and Validation of the Production Process.

The ileocolonic-targeted delivery of vitamins can establish beneficial alterations in gut microbial composition. Here, we describe the development of capsules containing riboflavin, nicotinic acid, and ascorbic acid covered with a pH-sensitive coating (ColoVit) to establish site-specific release in the ileocolon. Ingredient properties (particle size distribution, morphology) relevant for formulation and product quality were determined. Capsule content and the in vitro release behaviour were determined using a HPLC-method. Uncoated and coated validation batches were produced. Release characteristics were evaluated using a gastro-intestinal simulation system. All capsules met the required specifications. The contents of the ingredients were in the 90.0-120.0% range, and uniformity requirements were met. In the dissolution test a lag-time in drug release of 277-283 min was found, which meets requirements for ileocolonic release. The release itself is immediate as shown by dissolution of the vitamins of more than 75% in 1 h. The production process of the ColoVit formulation was validated and reproducible, it was shown that the vitamin blend was stable during the production process and in the finished coated product. The ColoVit is intended as an innovative treatment approach for beneficial microbiome modulation and optimization of gut health.

An in silico approach to unveil peptides from Acheta domesticus with potential bioactivity against hypertension, diabetes, cardiac and pulmonary fibrosis

Entomophagy is a sustainable alternative source of proteins for human nutrition. Acheta domesticus is one of the three insect species that complies with the European Union Regulation on novel foods, but to date, there are no reports on their potential bioactive peptides. In this study, an in silico approach was applied to simulate the gastrointestinal (GI) digestion of six A. domesticus proteins and identify new peptides with potential anti-hypertensive and/or anti-diabetic properties, resulting from their capability to inhibit the somatic Angiotensin-I converting enzyme (sACE) and/or dipeptidyl peptidase 4 (DPP-4), respectively. A molecular docking protocol was applied to evaluate the binding interactions between the 43 peptides ranked with high probability of being bioactive and three drug targets: DPP-4 and two catalytic domains (N- and C-) of sACE. Five peptides (AVQPCF, CAIAW, IIIGW, DATW and QIVW) showed high docking scores for both enzymes, suggesting their potential to inhibit the DPP-4 and both catalytic domains of sACE, thus possessing multifunctional bioactive properties. Two peptides (PIVCF and DVW) showed higher docking scores for the N-domain of sACE, indicating a potential action as selective inhibitors and consequently with anti-cardiac and pulmonary fibrosis bioactivities. This is the first study identifying peptides originated from the simulated GI digestion of A. domesticus with potential activities against hypertension, diabetes, cardiac and pulmonary fibrosis.

Effect of fiber-bound polyphenols from highland barley on lipid oxidation products of cooked pork during in vitro gastrointestinal digestion.

The gastrointestinal (GI) tract is a major site of lipid oxidation, and the lipid oxidation products are related to an increased risk of various chronic diseases. In this study, the inhibition capacity of bound-polyphenol rich insoluble dietary fiber (BP-IDF) from highland barley (HB) to lipid oxidation was evaluated during simulated GI digestion. We found that the level of lipid hydroperoxides (LOOH) and aldehydes were significantly inhibited when highland barley bound-polyphenol rich insoluble dietary fiber (HBBP-IDF) co-digestion with cooked pork. The lipid oxidation products were more effectively scavenged during simulated gastric digestion, with inhibition of 77.4% for LOOH, 52.3% for malondialdehyde, 46.5% for 4-hydroxy-2-hexenal and 48.7% for 4-hydroxy-2-nonenel, respectively. The fiber-bound polyphenols are the principal scavengers of lipid oxidation products. These findings suggest that HBBP-IDF could be used as a functional ingredient able to scavenge lipid oxidation products across the GI tract. © 2023 Society of Chemical Industry.

Effect of Food and a Proton-Pump Inhibitor on the Absorption of Encorafenib: An In Vivo-In Vitro-In Silico Approach.

Encorafenib is a kinase inhibitor indicated for the treatment of patients with BRAF mutant melanoma and BRAF mutant metastatic colorectal cancer. To understand the effect of food and coadministration with a proton-pump inhibitor (PPI), in vitro, in vivo, and in silico data were generated to optimize the clinical dose, evaluate safety, and better understand the oral absorption process under these conditions. Study 1 evaluated the effect of food on the plasma pharmacokinetics, safety, and tolerability after a single oral dose of encorafenib 100 mg. Study 2 evaluated the same end points with coadministration of encorafenib and rabeprazole (PPI perpetrator). The in vitro gastrointestinal TIM-1 model was used to investigate the release of encorafenib and the amount available for absorption under different testing conditions (fasted, fed, and with the use of a PPI). The fasted, fed, and PPI states were predicted for the encorafenib commercial capsule in GastroPlus 9.8. In study 1, both AUCinf and AUClast decreased by 4% with the administration of a high-fat meal. The Cmax was 36% lower than with fasted conditions. All 3 exposure parameters in study 2 (AUCinf, AUClast, and Cmax) had mean changes of <10% when encorafenib was coadministered with a PPI. Using the in vitro gastrointestinal simulator TIM-1, the model demonstrated a similar release of drug, as the bioaccessible fraction, in the 3 conditions was equal (≥80%), predicting no PPI or food effect for this drug formulation. The modeling in GastroPlus 9.8 demonstrated complete absorption of encorafenib when formulated as an amorphous solid dispersion. To obtain these results, it was crucial to integrate the amorphous solubility of the drug that shows a 20-fold higher solubility at pH 6.8 compared with crystalline solubility. The increased amorphous solubility is likely the reason no PPI effect was observed compared with fasted state conditions. The prolongation in gastric emptying in the fed state resulted in delayed plasma Tmax for encorafenib. No dose adjustment is necessary when encorafenib is administered in the fed state or when coadministered with a PPI. Both the TIM-1 and physiologically based pharmacokinetic model results were consistent with the observed clinical data, suggesting that these will be valuable tools for future work.

Valorisation of Wasted Immature Tomato to Innovative Fermented Functional Foods

In this study, the lactic fermentation of immature tomatoes as a tool for food ingredient production was evaluated as a circular economy-oriented alternative for valorising industrial tomatoes that are unsuitable for processing and which have wasted away in large quantities in the field. Two lactic acid bacteria (LAB) were assessed as starter cultures in an immature tomato pulp fermentation to produce functional food ingredients with probiotic potential. The first trial evaluated the probiotic character of Lactiplantibacillus plantarum (LAB97, isolated from immature tomato microbiota) and Weissella paramesenteroides (C1090, from the INIAV collection) through in vitro gastrointestinal digestion simulation. The results showed that LAB97 and C1090 met the probiotic potential viability criterion by maintaining 6 log10 CFU/mL counts after in vitro simulation. The second trial assessed the LAB starters' fermentative ability. Partially decontaminated (110 °C/2 min) immature tomato pulp was used to prepare the individually inoculated samples (Id: LAB97 and C1090). Non-inoculated samples, both with and without thermal treatment (Id: CTR-TT and CTR-NTT, respectively), were prepared as the controls. Fermentation was undertaken (25 °C, 100 rpm) for 14 days. Throughout storage (0, 24, 48, 72 h, 7, and 14 days), all the samples were tested for LAB and Y&M counts, titratable acidity (TA), solid soluble content (SSC), total phenolic content (TPC), antioxidant capacity (AOx), as well as for organic acids and phenolic profiles, and CIELab colour and sensory evaluation (14th day). The LAB growth reached ca. 9 log10 CFU/mL for all samples after 72 h. The LAB97 samples had an earlier and higher acidification rate than the remaining ones, and they were highly correlated to lactic acid increments. The inoculated samples showed a faster and higher decrease rate in their SSC levels when compared to the controls. A nearly two-fold increase (p < 0.05) during the fermentation, over time, was observed in all samples' AOx and TPC (p < 0.05, r = 0.93; similar pattern). The LAB97 samples obtained the best sensory acceptance for flavour and overall appreciation scores when compared to the others. In conclusion, the L. plantarum LAB97 starter culture was selected as a novel probiotic candidate to obtain a potential probiotic ingredient from immature tomato fruits.

The Effect of Encapsulating a Prebiotic-Based Biopolymer Delivery System for Enhanced Probiotic Survival

Orally delivered probiotics must survive transit through harsh environments during gastrointestinal (GI) digestion and be delivered and released into the target site. The aim of this work was to evaluate the survivability and delivery of gel-encapsulated Lactobacillus rhamnosus GG (LGG) to the colon. New hybrid symbiotic beads alginate/prebiotic pullulan/probiotic LGG were obtained by the extrusion method. The average size of the developed beads was 3401 µm (wet), 921 µm (dry) and the bacterial titer was 109 CFU/g. The morphology of the beads was studied by a scanning electron microscope, demonstrating the structure of the bacterial cellulose shell and loading with probiotics. For the first time, we propose adding an enzymatic extract of feces to an artificial colon fluid, which mimics the total hydrolytic activity of the intestinal microbiota. The beads can be digested by fecalase with cellulase activity, indicating intestinal release. The encapsulation of LGG significantly enhanced their viability under simulated GI conditions. However, the beads, in combination with the prebiotic, provided greater protection of bacteria, enhancing their survival and even increasing cell numbers in the capsules. These data suggest the promising prospects of coencapsulation as an innovative delivery method based on the inclusion of probiotic bacteria in a symbiotic matrix.

Chitosan/Albumin Coating Factorial Optimization of Alginate/Dextran Sulfate Cores for Oral Delivery of Insulin.

The design of nanoparticle formulations composed of biopolymers, that govern the physicochemical properties of orally delivered insulin, relies on improving insulin stability and absorption through the intestinal mucosa while protecting it from harsh conditions in the gastrointestinal (GI) tract. Chitosan/polyethylene glycol (PEG) and albumin coating of alginate/dextran sulfate hydrogel cores are presented as a multilayer complex protecting insulin within the nanoparticle. This study aims to optimize a nanoparticle formulation by assessing the relationship between design parameters and experimental data using response surface methodology through a 3-factor 3-level optimization Box-Behnken design. While the selected independent variables were the concentrations of PEG, chitosan and albumin, the dependent variables were particle size, polydispersity index (PDI), zeta potential, and insulin release. Experimental results showed a nanoparticle size ranging from 313 to 585 nm, with PDI from 0.17 to 0.39 and zeta potential ranging from -29 to -44 mV. Insulin bioactivity was maintained in simulated GI media with over 45% cumulative release after 180 min in a simulated intestinal medium. Based on the experimental responses and according to the criteria of desirability on the experimental region's constraints, solutions of 0.03% PEG, 0.047% chitosan and 1.20% albumin provide an optimum nanoparticle formulation for insulin oral delivery.

Multi-Compartmental Dissolution Method, an Efficient Tool for the Development of Enhanced Bioavailability Formulations Containing Poorly Soluble Acidic Drugs

The purpose of this study was to investigate the applicability of the Gastrointestinal Simulator (GIS), a multi-compartmental dissolution model, to predict the in vivo performance of Biopharmaceutics Classification System (BCS) Class IIa compounds. As the bioavailability enhancement of poorly soluble drugs requires a thorough understanding of the desired formulation, the appropriate in vitro modelling of the absorption mechanism is essential. Four immediate release ibuprofen 200 mg formulations were tested in the GIS using fasted biorelevant media. In addition to the free acid form, ibuprofen was present as sodium and lysine salts in tablets and as a solution in soft-gelatin capsules. In the case of rapid-dissolving formulations, the dissolution results indicated supersaturation in the gastric compartment, which affected the resulting concentrations in the duodenum and the jejunum as well. In addition, a Level A in vitro-in vivo correlation (IVIVC) model was established using published in vivo data, and then the plasma concentration profiles of each formulation were simulated. The predicted pharmacokinetic parameters were consistent with the statistical output of the published clinical study. In conclusion, the GIS method was found to be superior compared to the traditional USP method. In the future, the method can be useful for formulation technologists to find the optimal technique to enhance the bioavailability of poorly soluble acidic drugs.

Microstructured lipid microparticles containing anthocyanins: Production, characterization, storage, and resistance to the gastrointestinal tract

Anthocyanins from grape peel extract have several biological properties and can act as a natural colorant and antioxidant agent. However, these compounds are susceptible to degradation by light, oxygen, temperature, and the gastrointestinal tract. Thus, this study produced microstructured lipid microparticles (MLMs) containing anthocyanins by the spray chilling technique and evaluated the particle stability. trans-free fully hydrogenated palm oil (FHPO) and palm oil (PO) were used as encapsulating materials in the ratios 90:10, 80:20, 70:30, 60:40, and 50:50, respectively. The concentration of grape peel extract was 40 % (w/w) in relation to the encapsulating materials. The microparticles were evaluated for thermal behavior by DSC, polymorphism, FTIR, size distribution and particle diameter, bulk density, tapped density, flow properties, morphology, phenolic compounds content, antioxidant capacity, and retention of anthocyanins. Furthermore, the storage stability of the microparticles was investigated at different temperatures (-18, 4, and 25 °C), and the anthocyanins retention capacity, kinetic parameters (half-life time and degradation constant rate), total color difference, and visual aspects were evaluated during 90 days of storage. The resistance of MLMs to the gastrointestinal tract was also evaluated. In general, higher FHPO concentrations increased the thermal resistance of the MLMs and both showed defined peaks of β' and β forms. The FTIR analysis showed that the MLMs preserved the original forms of their constituent materials even after atomization, with interactions between them. The increase in the PO concentration directly affected the increased mean particle diameter, agglomeration, and cohesiveness, as well as lower bulk density, tapped density, and flowability. The retention of anthocyanins in MLMs ranged from 81.5 to 61.3 % and was influenced by the particle size, with a better result observed for the treatment MLM_90:10. The same behavior was observed for the phenolic compounds content (1443.1–1247.2 mg GAE/100 g) and antioxidant capacity (1739.8–1660.6 mg TEAC/100 g). During the storage, MLMs made with FHPO to PO ratios of 80:20, 70:30, and 60:40 showed the highest stability for anthocyanin retention and color changes at the three temperatures (- 18 °C, 4 °C, and 25 °C). The gastrointestinal simulation in vitro revealed that all treatments were resistant to gastric phase and maintained a maximum and controlled release in the intestinal phase, demonstrating that FHPO together with PO are effective to protect anthocyanins during gastric digestion, and can improve the bioavailability of this compound in the human organism. Thus, the spray chilling technique may be a promising alternative for the production of anthocyanins-loaded microstructured lipid microparticles with functional properties for various technological applications.

Preparation, characterization and gastrointestinal stability of silk fibroin nanoparticles loaded with red wine polyphenols

Silk fibroin nanoparticles (SFN) are interesting drug delivery systems due to their high load capacity and low toxicity. The aim of this study was to reveal how SFN load red wine polyphenols (RWP), and to assess their protective effect during the early stages of digestion by using the dynamic gastrointestinal simulator (simgi®) under physiological conditions. RWP-SFN were prepared by incubation of SFN in red wine and recovered by centrifugation. The obtained nanoparticles (particle size of 194.4 ± 2.1 nm and a Zeta potential of −22.7 ± 0.1 mV) showed a significant polyphenol loading content (7386.59 ± 1018.28 μg/g) with a high encapsulation efficiency (66.66 ± 2.58%). They were highly stable during their passage through the digestive tract, being the activity of RWP highly preserved. This data paves the way for future developments of SFN as stabilizers with a wide range of applications in the field of food additives.

Potential Oral Anticancer Therapeutic Agents of Hexahydrocurcumin-Encapsulated Chitosan Nanoparticles against MDA-MB-231 Breast Cancer Cells.

Hexahydrocurcumin-encapsulated chitosan nanoparticles (HHC-CS-NPs) were formulated by oil-in-water emulsification and ionotropic gelation and optimized using the Box-Behnken design. The particle size, zeta potential, and encapsulation efficiency of the optimized HHC-CS-NPs were 256 ± 14 nm, 27.3 ± 0.7 mV, and 90.6 ± 1.7%, respectively. The TEM analysis showed a spherical shape and a dense structure with a narrow size distribution. The FT-IR analysis indicated no chemical interaction between the excipients and the drugs in the nanoparticles, but the existence of the drugs was molecularly dispersed in the nanoparticle matrices. The drug release profile showed a preliminary burst release followed by a sustained release under simulated gastrointestinal (GI) and physiological conditions. A stability study suggested that the HHC-CS-NPs were stable under UV light, simulated GI, and body fluids. The in vitro bioaccessibility and bioavailability of the HHC-CS-NPs were 2.2 and 6.1 times higher than those of the HHC solution, respectively. The in vitro evaluation of the antioxidant, anti-inflammatory, and cytotoxic effects of the optimized HHC-CS-NPs demonstrated that the CS-NPs significantly improved the biological activities of HHC in radical scavenging, hemolysis protection activity, anti-protein denaturation, and cytotoxicity against MDA-MB-231 breast cancer cells. Western blot analysis showed that the apoptotic protein expression of Bax, cytochrome C, caspase-3, and caspase-9, were significantly up-regulated, whereas the anti-apoptotic protein Bcl-2 expression was down-regulated in the HHC-CS-NP-treated cells. Our findings suggest that the optimized HHC-CS-NPs can be further developed as an efficient oral treatment for breast cancer.

Impact of gastrointestinal digestion simulation on brewer’s spent grain green extracts and their prebiotic activity

Brewer’s spent grain (BSG) is a by-product of the beer industry and a potential source of bioactive compounds. In this study, two methods of extracting bioactive compounds from brewer’s spent grain were used – solid-to-liquid conventional extraction (SLE) and solid-to-liquid ohmic heating extraction (OHE) coupled with two ratio combinations of solvents: 60 % and 80 % ethanol:water (v/v). The bioactive potential of the BSG extracts was assessed during the gastrointestinal tract digestion (GID) and the differences in their antioxidant activity, total phenolic content and characterization of the polyphenol profile was measured. The SLE extraction using 60 % ethanol:water (v/v) was the extraction method with higher antioxidant activity (33.88 mg ascorbic acid/g BSG – initial; 16.61 mg ascorbic acid/g BSG - mouth; 15.58 mg ascorbic acid/g BSG – stomach; 17.26 mg ascorbic acid/g BSG – duodenum) and higher content in total phenolics (13.26 mg gallic acid/g BSG – initial; 4.80 mg gallic acid/g BSG – mouth; 4.88 mg gallic acid/g BSG – stomach; 5.00 mg gallic acid/g BSG – duodenum). However, the OHE extraction using 80 % ethanol:water (v/v), had a higher bioaccessibility index (99.77 % for ferulic acid, 72.68 % for 4-hydroxybenzoic acid, 65.37 % for vanillin, 28.99 % for p-coumaric, 22.54 % for catechin) values of polyphenols. All the extracts enhanced (except for SLE for 60 % ethanol:water (v/v) at 2 and 1.5 %, and for 80 % ethanol:water (v/v) at 2 % with Bifidobacterium animalis spp. lactis BB12, where no growth was observed) the growth of the probiotic microorganisms tested (Bifidobacterium animalis B0 – O.D.’s between 0.8240 and 1.7727; Bifidobacterium animalis spp. lactis BB12 – O.D.’s between 0.7219 and 0.8798; Lacticaseibacillus casei 01 – O.D.’s between 0.9121 and 1.0249; and Lactobacillus acidophilus LA-5 – O.D.’s between 0.8595 and 0.9677), demonstrating a potential prebiotic activity of BSG extracts.