Intestinal Fluid Research Articles

Oxytocin Analogues for the Oral Treatment of Abdominal Pain.

Abdominal pain presents an onerous reality for millions of people affected by gastrointestinal disorders such as irritable bowel syndrome (IBS) and inflammatory bowel diseases (IBD). The oxytocin receptor (OTR) has emerged as a new analgesic drug target with OTR expression upregulated on colon-innervating nociceptors in chronic visceral hypersensitivity states, accessible via luminal delivery. However, the low gastrointestinal stability of OTR's endogenous peptide ligand oxytocin (OT) is a bottleneck for therapeutic development. Here, we report the development of potent and fully gut-stable OT analogues, laying the foundation for a new area of oral gut-specific peptide therapeutics. Ligand optimisation guided by structure-gut-stability-activity relationships yielded highly stable analogues (t1/2 >24 h, compared to t1/2 <10 min of OT in intestinal fluid) equipotent to OT (~3nM) and with enhanced OTR selectivity. Intra-colonic administration of the lead ligand significantly reduced colonic mechanical hypersensitivity in a concentration-dependent manner in a mouse model of chronic abdominal pain. Moreover, oral administration of the lead ligand also displayed significant analgesia in this abdominal pain mouse model. The generated ligands and employed strategies could pave the way to a new class of oral gut-specific peptides to study and combat chronic gastrointestinal disorders, an area with substantial unmet medical needs.

Studies on the Probiotic, Adhesion, and Induction Properties of Artisanal Lactic Acid Bacteria: to Customize a Gastrointestinal Niche to Trigger Anti-obesity Functions.

The primary goals of this work are to explore the potential of probiotic lactic acid bacteria's (LAB) mucin/mucus layer thickening properties and to identify anti-obesity candidate strains that improve appropriate habitat for use with the Akkermansia group population in the future. The HT-29 cell binding, antimicrobial properties, adhesion to the mucin/mucus layer, growth in the presence of mucin, stability during in vitro gastrointestinal (GI) conditions, biofilm formation, and mucin/mucus thickness increment abilities were all assessed for artisanal LAB strains. Sixteen LAB strains out of 40 were chosen for further analysis based on their ability to withstand GI conditions. Thirteen strains remained viable in simulated intestinal fluid, while most showed high viability in gastric juice simulation. Furthermore, 35.9-65.4% of those 16 bacteria adhered to the mucin layer. Besides, different lactate levels were produced, and Streptococcus thermophilus UIN9 exhibited the highest biofilm development. In the HT-29 cell culture, the highest mucin levels were 333.87µg/mL with O. AK8 at 50mM lactate, 313.38µg/mL with Lactobacillus acidophilus NRRL-B 1910 with initial mucin, and 311.41µg/mL with Lacticaseibacillus casei NRRL-B 441 with initial mucin and 50mM lactate. Nine LAB strains have been proposed as anti-obesity candidates, with olive isolates of Lactiplantibacillus plantarum being particularly important due to their ability to avoid mucin sugar consumption. Probiotic LAB's attachment to the colonic mucosa and its ability to stimulate HT-29 cells to secrete mucus are critical mechanisms that may support the development of Akkermansia.

Physicochemical and digestive properties of curcumin stabilized by quaternized chitosan/sodium alginate nanoemulsion coatings

Curcumin (Cur) is a bioactive substance, but its instability limits its use in the food system. Thus, a new polysaccharide-based coating is developed by combining quaternized chitosan (QCS) with sodium alginate (SA) in this study to improve the bioaccessibility and stability of Cur. The microstructure of the emulsion was characterized by fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and low-field pulsed nuclear magnetic resonance imaging (LF-NMR). The thermal, ionic strength, and pH stability of emulsions were measured. Results indicate that QCS/SA (1:1)-Cur is more stable than other three nanoemulsion systems and the encapsulation rate of which reaches 90.62 %. It also shows higher stability in gastric fluid, excellent bioaccessibility and FFAs in intestinal fluid, as well as significant antibacterial property, antioxidant activity and water dispersibility. This study introduces a new method for obtaining stable, functional Cur-coated systems for food applications.

A potential cocrystal strategy to tailor in-vitro dissolution and improve Caco-2 permeability and oral bioavailability of berberine

Berberine hydrochloride (BER), a promising candidate in treating tumors, diabetes and pain management, has relatively low oral absorption and bioavailability due to its low intestinal permeability. To address these challenges, we developed a BER and lornoxicam cocrystal (BLCC) by a solvent evaporation method and characterized it using X-ray diffraction, differential scanning calorimetry and thermogravimetric analysis. Compared with BER, BLCC exhibited an instant release in pH 1.0 HCl and a sustained release up to 24 h in pH 6.8 buffer solutions and water. The Caco-2 permeability of BLCC has shown a remarkable increase compared to that of BER (i.e., Papp(a→b): 50.30 × 10-7vs 8.82 × 10-7 cm/s), which is attributed to the improved lipophilicity of BER (i.e., log P: 1.29 vs −1.83) and the reduced efflux amount of BER (i.e., ER: 1.71 vs 12.11). Furthermore, BLCC demonstrated a relative bioavailability of 410 % in comparison to the original BER, due to notably enhanced intestinal permeability of BLCC and its continuous dissolution in simulated intestinal fluid. BLCC has the potential to tailor the dissolution behavior, improve intestinal permeability, and boost the bioavailability of BER. This indicates that the cocrystal strategy holds promise as an effective approach to improving the oral absorption and bioavailability of active pharmaceutical molecules with low permeability during drug development.

Garlic Bioconverted by Bacillus subtilis Stimulates the Intestinal Immune System and Modulates Gut Microbiota Composition.

This study evaluates the potential of bioconverted garlic ferments (BGFs) to stimulate the intestinal immune system and modulate cecal microbiota composition. In vitro, BGF significantly enhances Peyer's patch (PP)-mediated bone marrow cell proliferation and increases the production of interferon-gamma (IFN-γ), granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-6, and immunoglobulin A (IgA) but not IL-4, IL-5, and immunoglobulin E (IgE). Oral administration of BGF to C3H/HeN mice for 4 weeks significantly increases the GM-CSF (42.1-45.8 pgmL-1) and IFN-γ (6.5-12.1 pgmL-1) levels in PP cells. BGF also significantly elevates the levels of tumor necrosis factor-alpha (TNF-α, 165.0-236.3 pgmg-1), GM-CSF (2.4-3.0ngmg-1), and IFN-γ (1.5-3.2ngmg-1) in the small intestinal fluid, and TNF-α (2.2-3.1 pgmL-1) and IFN-γ (10.3-0.21.5 pgmL-1) in the mouse serum. Cecal microbial analysis reveals that BGF increases Bacteroidota and Verrucomicrobiota and decreases Actinobacteria and Bacillota at the phylum level in mice. At the genus level, BGF significantly increases the abundance of Fusimonas (250mgkg-1BW-1day-1), Bacteroides (125 and 250mgkg-1BW-1day-1), and Akkermansia (125mgkg-1BW-1day-1) and decreases that of Bifidobacterium (62.5 and 250mgkg-1BW-1day-1) and Limosilactobacillus (125 and 250mgkg-1BW-1day-1). This study provides the first evidence of BGF's ability to modulate the intestinal immune system and gut microbiota, supporting its potential as a novel functional material to enhance gut immunity.

Dissolution, phase behavior and mass transport of amorphous solid dispersions in aspirated human intestinal fluids

Amorphous solid dispersions (ASDs) typically show improved dissolution and generate supersaturated solutions, enhancing the oral bioavailability of poorly soluble drugs. To gain insights into intraluminal ASD behavior, we utilized two poorly soluble drugs with different crystallization tendencies, atazanavir and posaconazole, prepared as ASDs at a 10% drug loading with hydroxypropyl methylcellulose acetyl succinate (HPMCAS). We evaluated their release in aspirated fasted-state human intestinal fluid (FaHIF), and multi-component fasted-state simulated intestinal fluid (composite-FaSSIF), characterizing the supersaturation profiles and drug-rich nanodroplets that formed. Complete release was observed for atazanavir ASDs over a 90 min period. Flux for dissolved atazanavir ASDs remained high over the experimental time period of 3 h. In contrast, posaconazole solution concentrations were initially high and then decreased. Likewise, flux was initially high and then decreased where these changes are attributed to crystallization of the drug. Generation of spherical nano-sized amorphous droplets of ∼100-150 nm was found to occur in ex vivo FaHIF media for both ASDs, maximizing the diffusive flux during the supersaturation window. Moreover, buffer capacity differences were postulated to influence release rates of ASDs in simulated vs aspirated fluids. Importantly, the solution phase phenomena observed during ASD release in simulated fluids, namely amorphous nanodroplet formation and drug crystallization, were also found to occur in aspirated luminal fluids.

3D printing novel enteric capsule shells for personalized drug delivery

Individualized drug delivery presents a viable strategy for enhancing medication efficacy and patient safety. It involves producing small batches of custom dosages tailored to each patient’s individual needs. However, current pharmaceutical manufacturing processes are not designed for personalized dosage forms. Additive manufacturing processes have great potential in the pharmaceutical industry due to the customizable nature of 3D printing technologies and the increasing demand for customized pharmaceutics and drug delivery systems. Conventional enteric capsules are prepared by dip coating to achieve gastro-resistant release, but the manufacturing process is erratic and unreliable. This study aimed to investigate the feasibility of manufacturing enteric capsules from enteric polymers using the hot melt extrusion (HME) and fused filament fabrication (FFF) process. HME was utilized to prepare filaments using enteric polymers, Hypromellose acetate succinate (HPMC-AS), and Eudragit-L 100–55, with plasticizers, sorbitol, and polyethylene glycol 4000 (PEG-4000), and a thermoplastic polymer, polyvinyl alcohol (PVA). A computer-aided design program was utilized to design size 0 capsules. The prepared capsule design and filaments were used to fabricate capsules using the FFF process. Powdered red dye was used to simulate the drug and its release from FFF fabricated capsules was evaluated in simulated gastric and intestinal fluids, pH 1.2 and 6.8, respectively. Various combinations of enteric polymer, plasticizer, and thermoplastic polymer were examined. Thermogravimetric analysis (TGA) showed that the temperatures used for processing polymer blends were well below their thermal degradation temperatures, and dye release profiles were generated using image analysis software. This study demonstrates that capsules made with HPMCAS, PEG-4000, and PVA via the FFF process are suitable for controlling drug release and that the release profile can be modified by making small changes to the formulation.

Alginate sulfonamide hydrogel beads for 5-fluorouracil delivery: antitumor activity, cytotoxicity assessment, and theoretical investigation

This study focused on grafting a new monomer (E)-N-(4-(3-(4-bromophenyl) acryloyl) phenyl)-4-methyl benzene sulfonamide (Br-PS) onto sodium alginate (Alg) using a free radical polymerization method. The optimal parameters for the grafting polymerization reaction were investigated, including initiator and monomer concentrations, polymerization reaction duration, and temperature. Additionally, the conversion, graft, and solid content percentages were calculated. The resulting novel poly (Br-PS)-g-Alg was thoroughly analyzed using Fourier-transform infrared spectroscopy (FT-IR), proton nuclear magnetic resonance (1H NMR), and scanning electron microscopy (SEM). Moreover, poly (Br-PS)-g-Alg was tested for cytotoxicity and selectivity values on lung cancer cell line (A549), breast cancer cell line (MDA-MB-231), and a normal cell line (MDCK) using the neutral red uptake test. Poly (Br-PS)-g-Alg demonstrated more inhibitory impact (IC50 = 33.37 and 40.9 μg/mL) and high selectivity (selectivity index = 4.83 and 3.94) on the A549 and MDA-MB-231 cell lines, respectively. Furthermore, uniform beads of creative poly (Br-PS)-g-Alg were fabricated, and their swelling rate in various media was studied. These beads could potentially serve as drug carriers for 5-fluorouracil (5-FU). Release experiments in simulated gastric (SGF) and intestinal fluids (SIF) showed a slower 5-FU release pattern in SGF compared to SIF. The proposed structures of poly (Br-PS)-g-Alg were theoretically verified using density functional theory with DFT/B3LYP/6–31(G) basis set, revealing distinct interactions due to the presence of different functional groups. The findings of this study could significantly impact the development of new drug delivery systems.

Comparison of the characteristics of small intestinal fluid from white-feathered and yellow-feathered broilers

The objective of this study was to compare the properties of small intestinal fluid (SIF) from white- and yellow-feathered broilers fed variable dietary CP to test the hypothesis that digestive enzymes sourced from the SIF of yellow-feathered broilers could serve as a viable alternative for preparing in vitro SIF specifically for white-feathered broilers. Ten Arbor Acres Plus broilers (32 days of age) and 20 Chinese yellow chickens (51 days of age) were fitted with jejunal cannulas and assigned to dietary treatments as follows: 1) white-feathered broilers (BW of 4.08 ± 0.39 kg) fed diet 1 with CP of 19.88 %, 2) yellow-feathered broilers (BW of 2.89 ± 0.27 kg) fed diet 1, and 3) yellow-feathered broilers (BW of 2.84 ± 0.20 kg) fed diet 2 with CP of 16.32 %. Each treatment contained 10 replicates with 1 broiler in each replicate. Digestive enzyme activities, ion concentrations, pH, 16 amino acid (AA, excluding Tyr and Trp) contents, molecular weight distribution (MWD) of protein, hydrolysis rates on wheat starch, corn starch, casein, and soybean protein concentrate, and in vitro digestible energy of corn, soybean meal, and corn gluten meal were evaluated for SIF. Activities of chymotrypsin and amylase, pH, concentrations (mg/mL) of total amino acid (TAA) and 13 AA (excluding Pro, Cys, and Glu), and hydrolysis rates in casein or soybean protein concentrate were greater (P < 0.05), whereas MWD of protein from 19 to 23 kDa was lower (P < 0.05) in the SIF of yellow compared to white-feathered broilers fed diet 1. Furthermore, activities of chymotrypsin and amylase, concentrations of Na+, and Cl−, MWD of protein from 15 to 18 kDa were greater, while concentrations of K+ were lower in the SIF of yellow-feathered broilers fed diet 2 compared to white feathered broilers fed diet 1. Activities of chymotrypsin and amylase, K+ concentration, pH, TAA and 13 AA (excluding Pro, Cys, and Glu), hydrolysis rates in casein or soybean protein concentrate increased (P < 0.05), while concentrations of Na+ and Cl− (P < 0.05) decreased with dietary CP in SIF from yellow-feathered broilers. This experiment demonstrated diet composition and breed modulate digestive enzyme activities, ions (Na+, K+, Cl−), pH, and AA concentrations in the SIF. Our findings revealed no substantial difference in the hydrolytic characteristics of SIF from white-feathered broilers fed diet 1 and yellow-feathered broilers fed diet 2. Consequently, it is feasible to use digestive enzymes extracted from the SIF of yellow-feathered broilers to prepare simulated SIF for in vitro digestion of white-feathered broilers.

Delivery of imiquimod to intestinal lymph nodes following oral administration

Intestinal lymph nodes are involved in the progression of colorectal cancer (CRC). Tumours suppress the activation of dendritic cells (DCs) in draining lymph nodes, diminishing anti-cancer immune response. Imiquimod (IMQ) facilitates DCs activation via toll-like receptor 7, suggesting that targeted delivery of IMQ to intestinal lymph nodes can improve the treatment of CRC. This study aims to enhance the delivery of IMQ to intestinal lymph nodes by a highly lipophilic prodrug approach. Amide prodrugs were synthesised by conjugating IMQ with saturated and unsaturated medium- to long-chain fatty acids. Their potential for intestinal lymphatic transport was assessed by their affinity to chylomicrons and solubility in long-chain triglycerides. Further selection of prodrug candidates was determined by resistance to enzymatic hydrolysis in intestinal lumen and release of IMQ in the lymphatics using fasting state simulated intestinal fluid supplemented with esterases, brush border enzyme vesicles and plasma. Key pharmacokinetic parameters and biodistribution in rats were assessed for the most promising compounds, prodrugs 5 and 8. The plasma concentration–time profile of IMQ following oral administration of the prodrugs was less erratic in comparison to the administration of unmodified IMQ. The lymph-to-plasma ratios of IMQ concentration increased 1.9- and 1.7-fold using prodrugs 5 and 8 in comparison to administration of unmodified IMQ, respectively. Importantly, the average concentration of IMQ in mesenteric lymph nodes (MLN) was 11.2- and 7.6-fold higher than in plasma following the administration of prodrugs 5 and 8, respectively. Additionally, the non-specific wide distribution of IMQ into various organs and tissues was reduced with prodrugs. This work suggests that the highly lipophilic prodrug approach can efficiently deliver IMQ to intestinal lymphatics. In addition, this study demonstrates the feasibility of an amide prodrug approach for intestinal lymphatic targeting.

Effect on heat treatment on the physical stability, interfacial tension and in vitro digestion of whey protein-stabilized ω-6/ω-3 fatty acid balanced pumpkin seed oil complex condensate

Despite the fact that whey proteins (WP) have shown diverse functional and nutritional properties, they exhibit poor stability when exposed to temperatures above the thermal denaturation point. Therefore, in this study, a complex coacervation layer of formed by WP treated at different heating temperatures (55, 65, 75 and 85 °C) and gum arabic (GA) was used as a wall material to microencapsulate ω-6/ω-3 fatty acid balanced pumpkin seed oil (MPO) with a view to exploring the effect of heat treatment temperatures on the nature of microencapsulation stabilized by WP-GA complexes. At pH 3.6, WP and GA formed a strong electrostatic complex. When the heat treatment temperature reached 65 °C, the WP-GA complexes exhibited more excellent particle size, absolute potential, emulsification properties and structural changes, which made them more suitable as hydrophilic emulsifiers to stabilize oil/water (O/W) systems. Further research into the physical properties and interfacial characteristics of O/W emulsions formed by the WP-GA complex was conducted. When subjected to a heat treatment temperature of 65 °C, the O/W emulsions achieved a higher interfacial protein content (3.17 ± 0.08 mg/m2), along with a lower interfacial tension (15.52 mN/m). Additionally, the encapsulated MPO demonstrates superior oxidative stability compared to its unencapsulated counterpart. In-vitro simulated digestion experiments revealed that only small fraction (17.70 ± 1.13%) of the encapsulated oil was released in simulated gastric fluid, while the majority (76.70 ± 1.47%) was released in simulated intestinal fluid, which suggested that the WP-GA composite coacervates was a promising encapsulation shell material, capable of maintaining integrity in the gastric phase and effectively delivering the encapsulant to the intestinal phase.

Multifunctional nanoplatforms based on alumina-coated mesoporous silica with potential for cancer theranostics applications

Multifunctional nanoplatforms based on mesoporous silica coated with alumina were successfully and sustainably synthesized using sodium silicate extracted from rice husk ash (RHA), demonstrating the potential for cancer theranostics. The hybrid nanostructures produced (C2-600 and C4-600), from two different calcination times, exhibited distinct morphologies, textural parameters, and degrees of mesoscopic organization. As expected, the Al2O3 coating on the mesoporous silica nanoparticles (MSNs) reduced the specific surface area from 720 m2/g to 122 m2/g (C2-600) and 57 m2/g (C4-600), while preserving their mesoporous structure. Additionally, both C2-600 and C4-600 showed relatively good stability across a wide pH interval, with a zeta potential of ζ = -15 mV and hydrodynamic diameter (Dh) ranging from 160 to 670 nm, depending on the pH of the medium. These peculiar characteristics resulted in high encapsulation efficiency (EE ≈ 90%) for the doxorubicin (DOX) anticancer drug, as well as sustained drug release in simulated gastric fluid (SGF, pH 1.2) and simulated intestinal fluid (SIF, pH 7.4). Appreciably, C4-600-DOX released approximately 83% of the drug over 96 hours and demonstrated significant biodegradation in simulated biological media. Furthermore, the hybrid nanoplatforms exhibited strong optical absorption between 250 and 420 nm, along with broad and intense photoluminescence (PL) in the near-infrared (NIR) region (680-900 nm), which is highly desirable for NIR-fluorescence diagnostic imaging. Notably, the hybrid nanoplatforms without DOX were non-cytotoxic to fibroblast and Caco-2 cells, while the DOX-loaded nanoplatforms exhibited selectivity and potent anticancer activity, inhibiting approximately 80% of Caco-2 colorectal cancer cells after 72 hours. These findings demonstrate that C2-600 and C4-600 are innovative, multifunctional and biodegradable nanoplatforms with powerful potential as drug carriers and fluorescence imaging agents, making them promising candidates for cancer theranostics.

Interfacial behaviour of human bile and its substitution for in vitro lipolysis studies

This study examined the interfacial evolution of individual bile salts (BSs) and their blends with phosphatidylcholine (BS/PC) to simulate the complex behaviour of human bile (HB) during lipolysis at the triglyceride/water interface. Using adsorption and desorption cycles, mimicking exposure to small intestinal fluids, we demonstrate that the interfacial behaviour of HB can be replicated using simple mixtures of BSs and PC. Interfacial tension (IFT) measurements after lipolysis and desorption showed no significant differences (P > 0.05) between HB samples and BS/PC mixtures across the total BS concentrations analysed (2.23–7.81 mM). However, individual BSs without PC yielded significantly different IFT results (P < 0.01) compared to HB, highlighting the importance of phospholipids. Dilatation rheology further emphasised the need for accurate phospholipid representation in bile models. Our results suggest that phospholipids in HB and BS/PC systems enhance resistance to desorption, potentially affecting lipolysis. This is important, as current in vitro digestion models often replicate only intestinal BS concentrations to mimic the behaviour HB in the intestinal lumen. Furthermore, the specific composition of BSs in HB appears less critical than the overall BS and phospholipid content, suggesting that the kinetics of triglyceride digestion is influenced by the combined luminal concentrations of these components. These findings have significant implications for understanding the role of bile in digestion and offer insights for designing more accurate in vitro models to study food emulsions and lipid-based delivery systems.

Optimization of pH-responsive microgel for the co-delivery of Weizmannia coagulans and procyanidins to enhance survival rate and tolerance

The purpose of this study was to prepare a pH-responsive microgel for co-delivering Weizmannia coagulans 99 (BC99) and procyanidins (PCs) to enhance the survival rate and tolerance of probiotics in complex micro-environment. The effects of different concentrations of PCs on the properties of microgels were optimized, and found that the spherical microgels had higher encapsulation efficiency (90.27 ± 2.51 %) and smaller size when the concentration of PCs was 20 μg/mL. The interaction among PCs, pectin and protein could effectively improve the survival rate of BC99 under different pH, bile salt, digestive enzyme and temperature conditions, maintain their stability in acidic gastric fluid, and realize the release of probiotics in neutral intestinal fluid. Moreover, the microgel was able to protect BC99 against H2O2 and antibiotics. This work provides a pH-responsive co-loaded microgels for BC99 and PCs, and has the potential in the loading and delivery of other probiotics and polyphenols.

Preparation, Characterization and in Vitro Stability of Soy Protein Fibrils Ferrous Delivery System

Iron deficiency is considered widespread globally, and discovering ferrous ion delivery system with better stability is one of the keys to solving this problem. In this paper, a ferrous ion delivery system was prepared from soy isolate protein fibrils (SPNFs). The alterations in infrared spectrum and microstructure before and after loading were compared, and the ionic strength stability, freeze-thaw stability, and oxidative reaction activity of soy isolate protein fibrils loaded with ferrous iron (SPNFs-Fe) were analyzed, simulating the release kinetics of gastrointestinal digestion. The results demonstrated that SPNFs load ferrous ions with carboxyl, carbonyl, and amino groups. SPNFs-Fe had good ionic strength stability and excellent freeze-thaw stability and showed weak pro-oxidation activity. The release kinetics of SPNFs-Fe in simulated gastrointestinal digestion best fit the Ritger-Peppas model, which had a slow-release effect in simulated gastrointestinal digestion. The release of SPNFs-Fe in simulated gastric fluid belonged to Fick diffusion, while in simulated intestinal fluid it belonged to non-Fick diffusion. These results suggest that SPNFs-Fe is a promising ferrous delivery system as a supplementary source of iron.

Bionic cell wall models: Utilizing TEMPO-oxidized cellulose nanofibers for fucoxanthin delivery systems

Fucoxanthin (FX) has various excellent biological properties but suffers from poor bioavailability. In this work, we build a bionic cell wall model using TEMPO (2,2,6,6-tetramethyl-1-piperidinyloxy)-oxidized cellulose. The bionic cell wall enhances the environmental stability of the liposomes and serves as a pH-responsive mechanism. The coating processes protect the structure of liposomes and fucoxanthin under the acidic conditions of the stomach. The bionic cell wall dispersed and released the fucoxanthin in simulated intestinal fluid. Overall, the protective and release capabilities highlight the potential of cellulose in a bionic cell wall model and provide diversity for the structural design of carriers for delivering functional bioactive components.

Biomineralized ZIF-8 Encapsulating SOD from Hydrogenobacter Thermophilus: Maintaining Activity in the Intestine and Alleviating Intestinal Oxidative Stress.

Oxidative stress is a major factor leading to inflammation and disease occurrence, and superoxide dismutase (SOD) is a crucial antioxidative metalloenzyme capable of alleviating oxidative stress. In this study, a novel thermostable SOD gene is obtained from the Hydrogenobacter thermophilus strain (HtSOD), transformed and efficiently expressed in Escherichia coli with an activity of 3438 Umg-1, exhibiting excellent thermal stability suitable for scalable production. However, the activity of HtSOD is reduced to less than 10% under the acidic environment. To address the acid resistance and gastrointestinal stability issues, a biomimetic mineralization approach is employed to encapsulate HtSOD within the ZIF-8 (HtSOD@ZIF-8). Gastrointestinal simulation results show that HtSOD@ZIF-8 maintained 70% activity in simulated gastric fluid for 2h, subsequently recovering to 97% activity in simulated intestinal fluid. Cell and in vivo experiments indicated that HtSOD@ZIF-8 exhibited no cytotoxicity and do not impair growth performance. Furthermore, HtSOD@ZIF-8 increased the relative abundance of beneficial microbiota such as Dubosiella and Alistipes, mitigated oxonic stress and intestinal injury by reducing mitochondrial and total reactive oxygen species (ROS) levels in diquat-induced. Together, HtSOD@ZIF-8 maintains and elucidates activity in the intestine and biocompatibility, providing insights into alleviating oxidative stress in hosts and paving the way for scalable production.

A Polysaccharide-Based Oral-Vaccine Delivery System and Adjuvant for the Influenza Virus Vaccine.

Influenza virus enters the host body through the mucosal surface of the respiratory tract. An efficient immune response at the mucosal site can interfere with virus entry and prevent infection. However, formulating oral vaccines and eliciting an effective mucosal immune response including at respiratory mucosa presents numerous challenges including the potential degradation of antigens by acidic gastric fluids and the risk of antigen dilution and dispersion over a large surface area of the gut, resulting in minimal antigen uptake by the immune cells. Additionally, oral mucosal vaccines have to overcome immune tolerance in the gut. To address the above challenges, in the current study, we evaluated inulin acetate (InAc) nanoparticles (NPs) as a vaccine adjuvant and antigen delivery system for oral influenza vaccines. InAc was developed as the first polysaccharide polymer-based TLR4 agonist; when tailored as a nanoparticulate vaccine delivery system, it enhanced antigen delivery to dendritic cells and induced a strong cellular and humoral immune response. This study compared the efficacy of InAc-NPs as a delivery system for oral vaccines with Poly (lactic-co-glycolic acid) (PLGA) NPs, utilizing influenza A nucleoprotein (Inf-A) as an antigen. InAc-NPs effectively protected the encapsulated antigen in both simulated gastric (pH 1.1) and intestinal fluids (pH 6.8). Moreover, InAc-NPs facilitated enhanced antigen delivery to macrophages, compared to PLGA-NPs. Oral vaccination studies in Balb/c mice revealed that InAc-Inf-A NPs significantly boosted the levels of Influenza virus-specific IgG and IgA in serum, as well as total and virus-specific IgA in the intestines and lungs. Furthermore, mice vaccinated with InAc-Inf-A-NPs exhibited notably higher hemagglutination inhibition (HI) titers at mucosal sites compared to those receiving the antigen alone. Overall, our study underscores the efficacy of InAc-NPs in safeguarding vaccine antigens post-oral administration, enhancing antigen delivery to antigen-presenting cells, and eliciting higher virus-neutralizing antibodies at mucosal sites following vaccination.

Characterization and potential application of microspheres from sodium alginate cross-linked with pectin from Citrus depressa Hayata’s peels

BackgroundPectin from Taiwan Citrus depressa Hayata’s peels (CDH pectin) and sodium alginate (Na alginate) were mixed in neutral acidity to produce microhydrogel beads or microspheres. The potential use of the microspheres such as encapsulation materials for quercetin and nobiletin, DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging activity, toxic elements absorption ability, and thermal characteristics were explored.ResultsDifferent ratios of CDH pectin and Na alginate produced microspheres of varying sizes and shapes. The highest yield (47.59%) with the broadest diameter was obtained at a Na alginate—CDH pectin ratio of 2:1, while the smallest yield was obtained from Na alginate—CDH pectin ratio of 1:3 (24.13%). Increasing the amount of Na alginate resulted in more spherical microspheres, higher heavy metals (cobalt and nickel) removal rates, yet a lower swelling ratio. A high pectin concentration also increased the encapsulation efficiency of quercetin and nobiletin, reaching 91.5% and 86.74%, respectively. Quercetin and nobiletin release analysis (in vitro) showed a slow release of drugs from the microspheres. Less than 20% quercetin and nobiletin were released from the microspheres in SGF (simulated gastric fluid) pH 1.2 solution after 2 h and more than 40% of the encapsulated drug was released in SIF (simulated intestinal fluid) pH 6.8 after 4 h. The strong DPPH scavenging activity of quercetin (99%) was not hindered by encapsulation materials. ICP-OES (inductively coupled plasma–optical emission spectrometry) analysis demonstrated that the biopolymer can absorb cobalt and nickel from water. Thermogravimetric analysis (TGA) result showed that the combination of CDH pectin and Na alginate produced a biopolymer that exhibited a weight loss of only 1.86–4.33% at 100 °C.ConclusionsThese findings suggest that microspheres produced from CDH pectin cross-linked with sodium alginate had potential in nobiletin and quercetin encapsulation. Moreover, the polymer could absorb heavy metals and exhibit an important characteristic for hot food and beverage packaging applications.

The Use of Organoclays as Excipient for Metformin Delivery: Experimental and Computational Study

This work combines experimental and computational modeling studies for the preparation of a composite of metformin and an organoclay, examining the advantages of a Tunisian clay used for drug delivery applications. The clay mineral studied is a montmorillonite-like smectite (Sm-Na), and the organoclay derivative (HDTMA-Sm) was used as a drug carrier for metformin hydrochloride (MET). In order to assess the MET loading into the clays, these materials were characterized by means of cation exchange capacity assessment, specific surface area measurement, and with the techniques of X-ray diffraction (XRD), differential scanning calorimetry, X-ray fluorescence spectroscopy, and Fourier-transformed infrared spectroscopy. Computational molecular modeling studies showed the surface adsorption process, identifying the clay–drug interactions through hydrogen bonds, and assessing electrostatic interactions for the hybrid MET/Sm-Na and hydrophobic interactions and cation exchange for the hybrid MET/HDTMA-Sm. The results show that the clays (Sm-Na and HDTMA-Sm) are capable of adsorbing MET, reaching a maximum load of 12.42 and 21.97 %, respectively. The adsorption isotherms were fitted by the Freundlich model, indicating heterogeneous adsorption of the studied adsorbate–adsorbent system, and they followed pseudo-second-order kinetics. The calculations of ΔGº indicate the spontaneous and reversible nature of the adsorption. The calculation of ΔH° indicates physical adsorption for the purified clay (Sm-Na) and chemical adsorption for the modified clay (HDTMA-Sm). The release of intercalated MET was studied in media simulating gastric and intestinal fluids, revealing that the purified clay (Sm-Na) and the modified organoclay (HDTMA-Sm) can be used as carriers in controlled drug delivery in future clinical applications. The molecular modeling studies confirmed the experimental phenomena, showing that the main adsorption mechanism is the cation exchange process between proton and MET cations into the interlayer space.