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1.
Int J Mol Sci ; 25(9)2024 Apr 23.
Article En | MEDLINE | ID: mdl-38731809

Polysaccharide-based systems have very good emulsifying and stabilizing properties, and starch plays a leading role. Their modifications should add new quality features to the product to such an extent that preserves the structure-forming properties of native starch. The aim of this manuscript was to examine the physicochemical characteristics of the combinations of starch with phospholipids or lysozymes and determine the effect of starch modification (surface hydrophobization or biological additives) and preparation temperature (before and after gelatinization). Changes in electrokinetic potential (zeta), effective diameter, and size distribution as a function of time were analyzed using the dynamic light scattering and microelectrophoresis techniques. The wettability of starch-coated glass plates before and after modification was checked by the advancing and receding contact angle measurements, as well as the angle hysteresis, using the settle drop method as a complement to profilometry and FTIR. It can be generalized that starch dispersions are more stable than analogous n-alkane/starch emulsions at room and physiological temperatures. On the other hand, the contact angle hysteresis values usually decrease with temperature increase, pointing to a more homogeneous surface, and the hydrophobization effect decreases vs. the thickness of the substrate. Surface hydrophobization of starch carried out using an n-alkane film does not change its bulk properties and leads to improvement of its mechanical and functional properties. The obtained specific starch-based hybrid systems, characterized in detail by switchable wettability, give the possibility to determine the energetic state of the starch surface and understand the strength and specificity of interactions with substances of different polarities in biological processes and their applicability for multidirectional use.


Polysaccharides , Starch , Wettability , Starch/chemistry , Polysaccharides/chemistry , Temperature , Muramidase/chemistry , Hydrophobic and Hydrophilic Interactions , Phospholipids/chemistry , Chemical Phenomena , Emulsions/chemistry
2.
Int J Mol Sci ; 25(9)2024 Apr 25.
Article En | MEDLINE | ID: mdl-38731895

UVB radiation is known to induce photodamage to the skin, disrupt the skin barrier, elicit cutaneous inflammation, and accelerate the aging process. Agaricus blazei Murill (ABM) is an edible medicinal and nutritional fungus. One of its constituents, Agaricus blazei Murill polysaccharide (ABP), has been reported to exhibit antioxidant, anti-inflammatory, anti-tumor, and immunomodulatory effects, which suggests potential effects that protect against photodamage. In this study, a UVB-induced photodamage HaCaT model was established to investigate the potential reparative effects of ABP and its two constituents (A1 and A2). Firstly, two purified polysaccharides, A1 and A2, were obtained by DEAE-52 cellulose column chromatography, and their physical properties and chemical structures were studied. A1 and A2 exhibited a network-like microstructure, with molecular weights of 1.5 × 104 Da and 6.5 × 104 Da, respectively. The effects of A1 and A2 on cell proliferation, the mitochondrial membrane potential, and inflammatory factors were also explored. The results show that A1 and A2 significantly promoted cell proliferation, enhanced the mitochondrial membrane potential, suppressed the expression of inflammatory factors interleukin-1ß (IL-1ß), interleukin-8 (IL-8), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α), and increased the relative content of filaggrin (FLG) and aquaporin-3 (AQP3). The down-regulated JAK-STAT signaling pathway was found to play a role in the response to photodamage. These findings underscore the potential of ABP to ameliorate UVB-induced skin damage.


Agaricus , Cell Proliferation , Filaggrin Proteins , HaCaT Cells , Ultraviolet Rays , Agaricus/chemistry , Humans , Ultraviolet Rays/adverse effects , Cell Proliferation/drug effects , Membrane Potential, Mitochondrial/drug effects , Fungal Polysaccharides/pharmacology , Fungal Polysaccharides/chemistry , Polysaccharides/pharmacology , Polysaccharides/chemistry , Keratinocytes/drug effects , Keratinocytes/metabolism , Keratinocytes/radiation effects , Cytokines/metabolism
3.
Food Res Int ; 186: 114371, 2024 Jun.
Article En | MEDLINE | ID: mdl-38729729

In this study, the impact of soy hull polysaccharide (SHP) concentration on high-internal-phase emulsions (HIPEs) formation and the gastrointestinal viability of Lactobacillus plantarum within HIPEs were demonstrated. Following the addition of SHP, competitive adsorption with soy protein isolate (SPI) occurred, leading to increased protein adhesion to the oil-water interface and subsequent coating of oil droplets. This process augmented viscosity and enhanced HIPEs stability. Specifically, 1.8 % SHP had the best encapsulation efficiency and delivery efficiency, reaching 99.3 % and 71.1 %, respectively. After 14 d of continuous zebrafishs feeding, viable counts of Lactobacillus plantarum and complex probiotics in the intestinal tract was 1.1 × 107, 1.3 × 107, respectively. In vitro experiments further proved that HIPEs' ability to significantly enhance probiotics' intestinal colonization and provided targeted release for colon-specific delivery. These results provided a promising strategy for HIPEs-encapsulated probiotic delivery systems in oral food applications.


Emulsions , Lactobacillus plantarum , Polysaccharides , Probiotics , Soybean Proteins , Zebrafish , Soybean Proteins/chemistry , Animals , Polysaccharides/chemistry , Lactobacillus plantarum/metabolism , Glycine max/chemistry , Viscosity
4.
J Agric Food Chem ; 72(19): 10923-10935, 2024 May 15.
Article En | MEDLINE | ID: mdl-38691832

This study aimed to explore the ameliorative effects and potential mechanisms of Huangshan Umbilicaria esculenta polysaccharide (UEP) in dextran sulfate sodium-induced acute ulcerative colitis (UC) and UC secondary liver injury (SLI). Results showed that UEP could ameliorate both colon and liver pathologic injuries, upregulate mouse intestinal tight junction proteins (TJs) and MUC2 expression, and reduce LPS exposure, thereby attenuating the effects of the gut-liver axis. Importantly, UEP significantly downregulated the secretion levels of TNF-α, IL-1ß, and IL-6 through inhibition of the NF-κB pathway and activated the Nrf2 signaling pathway to increase the expression levels of SOD and GSH-Px. In vitro, UEP inhibited the LPS-induced phosphorylation of NF-κB P65 and promoted nuclear translocation of Nrf2 in RAW264.7 cells. These results revealed that UEP ameliorated UC and SLI through NF-κB and Nrf2-mediated inflammation and oxidative stress. The study first investigated the anticolitis effect of UEP, suggesting its potential for the treatment of colitis and colitis-associated liver disease.


Colitis , Dextran Sulfate , NF-E2-Related Factor 2 , NF-kappa B , Polysaccharides , Animals , Mice , Polysaccharides/pharmacology , Polysaccharides/chemistry , Polysaccharides/administration & dosage , Dextran Sulfate/adverse effects , Male , NF-E2-Related Factor 2/metabolism , NF-E2-Related Factor 2/genetics , Humans , Colitis/drug therapy , Colitis/chemically induced , Colitis/metabolism , RAW 264.7 Cells , NF-kappa B/metabolism , NF-kappa B/genetics , Mice, Inbred C57BL , Protective Agents/pharmacology , Protective Agents/administration & dosage , Protective Agents/chemistry , Liver/drug effects , Liver/metabolism , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , Tumor Necrosis Factor-alpha/immunology , Oxidative Stress/drug effects , Interleukin-1beta/genetics , Interleukin-1beta/metabolism , Interleukin-1beta/immunology , Interleukin-6/genetics , Interleukin-6/metabolism , Interleukin-6/immunology , Colitis, Ulcerative/drug therapy , Colitis, Ulcerative/metabolism , Colitis, Ulcerative/chemically induced , Chemical and Drug Induced Liver Injury/drug therapy , Chemical and Drug Induced Liver Injury/metabolism , Mucin-2/genetics , Mucin-2/metabolism
5.
Org Biomol Chem ; 22(19): 3986-3994, 2024 May 15.
Article En | MEDLINE | ID: mdl-38695061

Algae-based marine carbohydrate drugs are typically decorated with negative ion groups such as carboxylate and sulfate groups. However, the precise synthesis of highly sulfated alginates is challenging, thus impeding their structure-activity relationship studies. Herein we achieve a microwave-assisted synthesis of a range of highly sulfated mannuronate glycans with up to 17 sulfation sites by overcoming the incomplete sulfation due to the electrostatic repulsion of crowded polyanionic groups. Although the partially sulfated tetrasaccharide had the highest affinity for the receptor binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant, the fully sulfated octasaccharide showed the most potent interference with the binding of the RBD to angiotensin-converting enzyme 2 (ACE2) and Vero E6 cells, indicating that the sulfated oligosaccharides might inhibit the RBD binding to ACE2 in a length-dependent manner.


Angiotensin-Converting Enzyme 2 , Antiviral Agents , Microwaves , Polysaccharides , SARS-CoV-2 , SARS-CoV-2/drug effects , Antiviral Agents/pharmacology , Antiviral Agents/chemical synthesis , Antiviral Agents/chemistry , Chlorocebus aethiops , Angiotensin-Converting Enzyme 2/metabolism , Angiotensin-Converting Enzyme 2/antagonists & inhibitors , Angiotensin-Converting Enzyme 2/chemistry , Vero Cells , Polysaccharides/chemistry , Polysaccharides/pharmacology , Polysaccharides/chemical synthesis , Humans , Animals , Spike Glycoprotein, Coronavirus/antagonists & inhibitors , Spike Glycoprotein, Coronavirus/metabolism , Spike Glycoprotein, Coronavirus/chemistry , Hexuronic Acids/chemistry , Hexuronic Acids/pharmacology , Hexuronic Acids/chemical synthesis , Sulfates/chemistry , Sulfates/pharmacology , Sulfates/chemical synthesis , COVID-19 Drug Treatment , Structure-Activity Relationship
6.
J Agric Food Chem ; 72(19): 11140-11152, 2024 May 15.
Article En | MEDLINE | ID: mdl-38703140

Recently, oral deliverable strategies of multiple nutraceuticals for ulcerative colitis (UC) mitigation have attracted increasing attention. This study aimed to fabricate facile oral assemblies loaded with egg-white-derived peptides (EWDP) and curcumin based on carboxymethyl chitosan (CMCS) and an γ-cyclodextrin metal-organic framework (MOF). Herein, outer CMCS could coassemble with EWDP (both nutraceuticals and building blocks) into cobweb-like fibrils to promote bridging with inner MOF via coordinative noncovalent interactions (hydrogen bonding, hydrophobic interaction, and electrostatic interaction). Compared with conventional γ-cyclodextrin/MOF-based composites, the above coassembly could also endow the biocompatible assemblies with superior nanoscale colloidal properties, processing applicability (curcumin storage stability, bioaccessibility, and aqueous solubility), and bioactivity. Moreover, the oral synergism of EWDP and curcumin (initially nonsynergistic) for UC mitigation was achieved by alleviating inflammatory damage and gut microbiota imbalance. Overall, the novel assemblies could be a promising amplifier and platform to facilitate oral formulations of various nutraceuticals for food processing and UC relief.


Colitis, Ulcerative , Curcumin , Metal-Organic Frameworks , Peptides , Curcumin/chemistry , Curcumin/administration & dosage , Metal-Organic Frameworks/chemistry , Animals , Humans , Peptides/chemistry , Peptides/administration & dosage , Colitis, Ulcerative/drug therapy , Mice , Chitosan/chemistry , Egg White/chemistry , Polysaccharides/chemistry , Male , Administration, Oral , Drug Synergism , gamma-Cyclodextrins/chemistry , Drug Carriers/chemistry , Egg Proteins/chemistry
7.
Phys Chem Chem Phys ; 26(19): 14160-14170, 2024 May 15.
Article En | MEDLINE | ID: mdl-38712976

Protonated ions of fucose-containing oligosaccharides are prone to undergo internal glycan rearrangement which results in chimeric fragments that obfuscate mass-spectrometric analysis. Lack of accessible tools that would facilitate systematic analysis of glycans in the gas phase limits our understanding of this phenomenon. In this work, we use density functional theory modeling to interpret cryogenic IR spectra of Lewis a and blood group type H1 trisaccharides and to establish whether these trisaccharides undergo the rearrangement during gas-phase analysis. Structurally unconstrained search reveals that none of the parent ions constitute a thermodynamic global minimum. In contrast, predicted collision cross sections and anharmonic IR spectra provide a good match to available experimental data which allowed us to conclude that fucose migration does not occur in these antigens. By comparing the predicted structures with those obtained for Lewis x and blood group type H2 epitopes, we demonstrate that the availability of the mobile proton and a large difference in the relative stability of the parent ions and rearrangement products constitute the prerequisites for the rearrangement reaction.


Lewis Blood Group Antigens , Lewis Blood Group Antigens/chemistry , Epitopes/chemistry , Thermodynamics , Polysaccharides/chemistry , Density Functional Theory , Blood Group Antigens/chemistry , Spectrophotometry, Infrared , Oligosaccharides/chemistry , Trisaccharides/chemistry
8.
Carbohydr Polym ; 337: 122139, 2024 Aug 01.
Article En | MEDLINE | ID: mdl-38710550

A novel RG-I pectin-like polysaccharide, YJ3A1, was purified from the flowers of Rosa chinensis and its structure and hepatoprotective effect in vivo and in vitro were investigated. The backbone of this polysaccharide is mainly composed of 1, 4-galactan, 1, 4-linked α-GalpA and 1, 2-linked α-Rhap disaccharide repeating unit attached by 1, 6-linked ß-Galp or 1, 5-linked α-Araf on C-4 of the Rhap. Interestingly, oral administration of YJ3A1 significantly ameliorates NASH-associated inflammation, oxidative stress and fibrosis and does not affect the liver morphology of normal mice at a dose of 50 mg/kg. The mechanism study suggests that the biological activity may associate to inactivating of high-mobility group box 1 protein (HMGB1)/TLR4/NF-κB and Akt signaling pathways by restraining the expression and release of HMGB1, thereby impeding the effect of NASH. The current findings outline a novel leading polysaccharide for new drug candidate development against NASH.


HMGB1 Protein , NF-kappa B , Non-alcoholic Fatty Liver Disease , Pectins , Rosa , Signal Transduction , Toll-Like Receptor 4 , Animals , Rosa/chemistry , Toll-Like Receptor 4/metabolism , HMGB1 Protein/metabolism , NF-kappa B/metabolism , Non-alcoholic Fatty Liver Disease/drug therapy , Non-alcoholic Fatty Liver Disease/metabolism , Non-alcoholic Fatty Liver Disease/pathology , Signal Transduction/drug effects , Mice , Pectins/pharmacology , Pectins/chemistry , Pectins/isolation & purification , Male , Humans , Inflammation/drug therapy , Inflammation/metabolism , Mice, Inbred C57BL , Polysaccharides/pharmacology , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Oxidative Stress/drug effects
9.
Carbohydr Polym ; 337: 122171, 2024 Aug 01.
Article En | MEDLINE | ID: mdl-38710561

Commercially available mushroom polysaccharides have found widespread use as adjuvant tumor treatments. However, the bioactivity of polysaccharides in Lactarius hatsudake Tanaka (L. hatsudake), a mushroom with both edible and medicinal uses, remains relatively unexplored. To address this gap, five L. hatsudake polysaccharides with varying molecular weights were isolated, named LHP-1 (898 kDa), LHP-2 (677 kDa), LHP-3 (385 kDa), LHP-4 (20 kDa), and LHP-5 (4.9 kDa). Gas chromatography-mass spectrometry, nuclear magnetic resonance, and atomic force microscopy, etc., were employed to determine their structural characteristics. The results confirmed that spherical aggregates with amorphous flexible fiber chains dominated the conformation of the LHP. LHP-1 and LHP-2 were identified as glucans with α-(1,4)-Glcp as the main chain; LHP-3 and LHP-4 were classified as galactans with varying molecular weights but with α-(1,6)-Galp as the main chain; LHP-5 was a glucan with ß-(1,3)-Glcp as the main chain and ß-(1,6)-Glcp connecting to the side chains. Significant differences were observed in inhibiting tumor cell cytotoxicity and the antioxidant activity of the LHPs, with LHP-5 and LHP-4 identified as the principal bioactive components. These findings provide a theoretical foundation for the valuable use of L. hatsudake and emphasize the potential application of LHPs in therapeutic tumor treatments.


Antioxidants , Glucans , Glucans/chemistry , Glucans/pharmacology , Glucans/isolation & purification , Humans , Antioxidants/chemistry , Antioxidants/pharmacology , Antioxidants/isolation & purification , Agaricales/chemistry , Polysaccharides/chemistry , Polysaccharides/pharmacology , Polysaccharides/isolation & purification , Molecular Weight , Antineoplastic Agents/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/isolation & purification , Cell Line, Tumor , Fungal Polysaccharides/chemistry , Fungal Polysaccharides/pharmacology , Fungal Polysaccharides/isolation & purification , Basidiomycota/chemistry , Cell Survival/drug effects
10.
Carbohydr Polym ; 337: 122157, 2024 Aug 01.
Article En | MEDLINE | ID: mdl-38710573

Seaweed polysaccharides, particularly sulfated ones, exhibited potent antiviral activity against a wide variety of enveloped viruses, such as herpes simplex virus and respiratory viruses. Different mechanisms of action were suggested, which may range from preventing infection to intracellular antiviral activity, at different stages of the viral cycle. Herein, we generated two chemically engineered sulfated fucans (C303 and C304) from Cystoseira indica by an amalgamated extraction-sulfation procedure using chlorosulfonic acid-pyridine/N,N-dimethylformamide and sulfur trioxide-pyridine/N,N-dimethylformamide reagents, respectively. These compounds exhibited activity against HSV-1 and RSV with 50 % inhibitory concentration values in the range of 0.75-2.5 µg/mL and low cytotoxicity at concentrations up to 500 µg/mL. The antiviral activities of chemically sulfated fucans (C303 and C304) were higher than the water (C301) and CaCl2 extracted (C302) polysaccharides. Compound C303 had a (1,3)-linked fucan backbone and was branched. Sulfates were present at positions C-2, C-4, and C-2,4 of Fucp, and C-6 of Galp residues of this polymer. Compound C304 had a comparable structure but with more sulfates at C-4 of Fucp residue. Both C303 and C304 were potent antiviral candidates, acting in a dose-dependent manner on the adsorption and other intracellular stages of HSV-1 and RSV replication, in vitro.


Antiviral Agents , Herpesvirus 1, Human , Polysaccharides , Antiviral Agents/pharmacology , Antiviral Agents/chemistry , Chlorocebus aethiops , Herpesvirus 1, Human/drug effects , Polysaccharides/pharmacology , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Animals , Vero Cells , Humans , Sulfates/chemistry , Sulfates/pharmacology , Respiratory Syncytial Viruses/drug effects
11.
Carbohydr Polym ; 337: 122156, 2024 Aug 01.
Article En | MEDLINE | ID: mdl-38710572

Seaweeds represent a rich source of sulfated polysaccharides with similarity to heparan sulfate, a facilitator of myriad virus host cell attachment. For this reason, attention has been drawn to their antiviral activity, including the potential for anti-SARS-CoV-2 activity. We have identified and structurally characterized several fucoidan extracts, including those from different species of brown macroalga, and a rhamnan sulfate from a green macroalga species. A high molecular weight fucoidan extracted from Saccharina japonica (FSjRPI-27), and a rhamnan sulfate extracted from Monostroma nitidum (RSMn), showed potent competitive inhibition of spike glycoprotein receptor binding to a heparin-coated SPR chip. This inhibition was also observed in cell-based assays using hACE2 HEK-293 T cells infected by pseudotyped SARS-CoV-2 virus with IC50 values <1 µg/mL. Effectiveness was demonstrated in vivo using hACE2-transgenic mice. Intranasal administration of FSjRPI-27 showed protection when dosed 6 h prior to and at infection, and then every 2 days post-infection, with 100 % survival and no toxicity at 104 plaque-forming units per mouse vs. buffer control. At 5-fold higher virus dose, FSjRPI-27 reduced mortality and yielded reduced viral titers in bronchioalveolar fluid and lung homogenates vs. buffer control. These findings suggest the potential application of seaweed-based sulfated polysaccharides as promising anti-SARS-CoV-2 prophylactics.


Antiviral Agents , COVID-19 , Mannans , Polysaccharides , SARS-CoV-2 , Seaweed , Polysaccharides/chemistry , Polysaccharides/pharmacology , Animals , Humans , SARS-CoV-2/drug effects , Seaweed/chemistry , Antiviral Agents/pharmacology , Antiviral Agents/chemistry , HEK293 Cells , Mice , COVID-19/prevention & control , COVID-19/virology , COVID-19 Drug Treatment , Mice, Transgenic , Spike Glycoprotein, Coronavirus/metabolism , Deoxy Sugars/pharmacology , Deoxy Sugars/chemistry , Angiotensin-Converting Enzyme 2/metabolism
12.
J Mass Spectrom ; 59(6): e5034, 2024 Jun.
Article En | MEDLINE | ID: mdl-38726698

Glycosylation is an incredibly common and diverse post-translational modification that contributes widely to cellular health and disease. Mass spectrometry is the premier technique to study glycoproteins; however, glycoproteomics has lagged behind traditional proteomics due to the challenges associated with studying glycosylation. For instance, glycans dissociate by collision-based fragmentation, thus necessitating electron-based fragmentation for site-localization. The vast glycan heterogeneity leads to lower overall abundance of each glycopeptide, and often, ion suppression is observed. One of the biggest issues facing glycoproteomics is the lack of reliable software for analysis, which necessitates manual validation and serves as a massive bottleneck in data processing. Here, I will discuss each of these challenges and some ways in which the field is attempting to address them, along with perspectives on how I believe we should move forward.


Glycomics , Glycoproteins , Mass Spectrometry , Proteomics , Proteomics/methods , Glycomics/methods , Mass Spectrometry/methods , Glycoproteins/analysis , Glycoproteins/chemistry , Humans , Glycosylation , Polysaccharides/analysis , Polysaccharides/chemistry , Glycopeptides/analysis , Glycopeptides/chemistry , Software , Protein Processing, Post-Translational , Animals
13.
Cryo Letters ; 45(3): 149-157, 2024.
Article En | MEDLINE | ID: mdl-38709186

BACKGROUND: The industrial scale cryo-storage of raw tissue materials requires a robust, low-cost and easy-to-operate method that can facilitate the down-stream process. OBJECTIVE: The study was aimed to develop the multifunctional protective solutions (MPS) for transportation at ambient conditions and also subsequent cryo-storage below -20 degree C of raw porcine hides for tissue engineering and regenerative medicine. MATERIALS AND METHODS: Protective solutions with antimicrobial activity and proteinase-inhibiting activity were developed and tested for its efficacy in preserving the extracellular matrix of porcine dermis from microbial spoilage, proteolytic degradation, freeze damage and excessive dehydration during shipping and cryo-storage. The MPSs contained phosphate-buffered saline with ethylene diamine tetra acetic acid (EDTA) added as chelator and proteinase inhibitor, as well as glycerol or maltodextrin (M180) as cryoprotectants. RESULTS: MPSs prepared with EDTA and glycerol or M180 had significant antimicrobial activity and proteinase-inhibiting activity during the period of shipping and handling. Glycerol and M180 prevented eutectic salt precipitation and excessive freeze dehydration upon cryo-storage of porcine hides. Without glycerol or M180, hides could be freeze-dehydrated to the low hydration at ~0.4 g/g dw, and formed irreversible plications after freezing. A critical hydration (0.8~0.9 g/g dw) was observed for the extracellular matrix of porcine dermis, and dehydration to a lower level could impose enormous stress and potential damage. The soaking of porcine hides in MPSs decreased water content as glycerol and M180 entered into dermis. Upon equilibration, the glycerol content in the tissue was about 94% of the incubating glycerol solution, but the M180 content in the tissue was only about 50% of the incubating M180 solution, indicating that M180 did not get into the entire aqueous domain within dermis. MPSs reduced ice formation and increased the unfrozen water content of porcine raw hides upon cryo-storage. CONCLUSION: MPSs prepared with EDTA and glycerol or M180 have antimicrobial activity and proteinase-inhibiting activity, which can be used for transportation and cryo-storage of raw hides at the industrial scale. Glycerol at 7.5% w/v and M180 at 20% w/v were sufficient to prevent freeze damage and excessive freeze dehydration. Doi.org/10.54680/fr24310110312.


Cryopreservation , Cryoprotective Agents , Regenerative Medicine , Tissue Engineering , Animals , Regenerative Medicine/methods , Swine , Tissue Engineering/methods , Cryopreservation/methods , Cryoprotective Agents/pharmacology , Cryoprotective Agents/chemistry , Edetic Acid/chemistry , Edetic Acid/pharmacology , Protease Inhibitors/pharmacology , Protease Inhibitors/chemistry , Polysaccharides/chemistry , Polysaccharides/pharmacology , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Extracellular Matrix/chemistry , Extracellular Matrix/drug effects
14.
AAPS PharmSciTech ; 25(5): 99, 2024 May 07.
Article En | MEDLINE | ID: mdl-38714608

Hypericum perforatum (HP) contains valuable and beneficial bioactive compounds that have been used to treat or prevent several illnesses. Encapsulation technology offers protection of the active compounds and facilitates to expose of the biologically active compounds in a controlled mechanism. Microcapsulation of the hydroalcoholic gum arabic and maltodextrin have hot been used as wall materials in the encapsulation of HP extract. Therefore, the optimum microencapsulation parameters of Hypericum perforatum (HP) hydroalcoholic extract were determined using response surface methodology (RSM) for the evaluation of HP extract. Three levels of three independent variables were screened using the one-way ANOVA. Five responses were monitored, including total phenolic content (TPC), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), carr index (CI), hausner ratio (HR), and solubility. Optimum drying conditions for Hypericum perforatum microcapsules (HPMs) were determined: 180 °C for inlet air temperature, 1.04/1 for ratio of maltodextrin to gum arabic (w/w), and 1.98/1 for coating to core material ratio (w/w). TPC, antioxidant activity, CI, HR, and solubility values were specified as 316.531 (mg/g GAE), 81.912%, 6.074, 1.066, and 35.017%, respectively, under the optimized conditions. The major compounds of Hypericum perforatum (hypericin and pseudohypericin) extract were determined as 4.19 µg/g microcapsule and 15.09 µg/g microcapsule, respectively. Scanning electron microscope (SEM) analysis revealed that the mean particle diameter of the HPMs was 20.36 µm. Based on these results, microencapsulation of HPMs by spray drying is a viable technique which protects the bioactive compounds of HP leaves, facilitating its application in the pharmaceutical, cosmetic, and food industries.


Antioxidants , Capsules , Drug Compounding , Gum Arabic , Hypericum , Plant Extracts , Polysaccharides , Solubility , Hypericum/chemistry , Plant Extracts/chemistry , Drug Compounding/methods , Gum Arabic/chemistry , Polysaccharides/chemistry , Antioxidants/chemistry , Antioxidants/pharmacology , Capsules/chemistry , Spray Drying , Phenols/chemistry , Desiccation/methods
15.
AAPS PharmSciTech ; 25(5): 101, 2024 May 07.
Article En | MEDLINE | ID: mdl-38714629

BACKGROUND: Niacin, an established therapeutic for dyslipidemia, is hindered by its propensity to induce significant cutaneous flushing when administered orally in its unmodified state, thereby constraining its clinical utility. OBJECTIVE: This study aimed to fabricate, characterize, and assess the in-vitro and in-vivo effectiveness of niacin-loaded polymeric films (NLPFs) comprised of carboxymethyl tamarind seed polysaccharide. The primary objective was to mitigate the flushing-related side effects associated with oral niacin administration. METHODS: NLPFs were synthesized using the solvent casting method and subsequently subjected to characterization, including assessments of tensile strength, moisture uptake, thickness, and folding endurance. Surface characteristics were analyzed using a surface profiler and scanning electron microscopy (SEM). Potential interactions between niacin and the polysaccharide core were investigated through X-ray diffraction experiments (XRD) and Fourier transform infrared spectroscopy (FTIR). The viscoelastic properties of the films were explored using a Rheometer. In-vitro assessments included drug release studies, swelling behavior assays, and antioxidant assays. In-vivo efficacy was evaluated through skin permeation assays, skin irritation assays, and histopathological analyses. RESULTS: NLPFs exhibited a smooth texture with favorable tensile strength and moisture absorption capabilities. Niacin demonstrated interaction with the polysaccharide core, rendering the films amorphous. The films displayed slow and sustained drug release, exceptional antioxidant properties, optimal swelling behavior, and viscoelastic characteristics. Furthermore, the films exhibited biocompatibility and non-toxicity towards skin cells. CONCLUSION: NLPFs emerged as promising carrier systems for the therapeutic transdermal delivery of niacin, effectively mitigating its flushing-associated adverse effects.


Administration, Cutaneous , Drug Liberation , Niacin , Polysaccharides , Rats, Wistar , Skin Absorption , Skin , Animals , Rats , Niacin/administration & dosage , Niacin/chemistry , Niacin/pharmacology , Polysaccharides/chemistry , Polysaccharides/administration & dosage , Polysaccharides/pharmacology , Skin/metabolism , Skin/drug effects , Skin Absorption/drug effects , Flushing/chemically induced , Tensile Strength , Male , Drug Delivery Systems/methods , Tamarindus/chemistry , Polymers/chemistry
16.
Nat Commun ; 15(1): 3755, 2024 May 04.
Article En | MEDLINE | ID: mdl-38704385

Heparin is an important anticoagulant drug, and microbial heparin biosynthesis is a potential alternative to animal-derived heparin production. However, effectively using heparin synthesis enzymes faces challenges, especially with microbial recombinant expression of active heparan sulfate N-deacetylase/N-sulfotransferase. Here, we introduce the monosaccharide N-trifluoroacetylglucosamine into Escherichia coli K5 to facilitate sulfation modification. The Protein Repair One-Stop Service-Focused Rational Iterative Site-specific Mutagenesis (PROSS-FRISM) platform is used to enhance sulfotransferase efficiency, resulting in the engineered NST-M8 enzyme with significantly improved stability (11.32-fold) and activity (2.53-fold) compared to the wild-type N-sulfotransferase. This approach can be applied to engineering various sulfotransferases. The multienzyme cascade reaction enables the production of active heparin from bioengineered heparosan, demonstrating anti-FXa (246.09 IU/mg) and anti-FIIa (48.62 IU/mg) activities. This study offers insights into overcoming challenges in heparin synthesis and modification, paving the way for the future development of animal-free heparins using a cellular system-based semisynthetic strategy.


Anticoagulants , Escherichia coli , Heparin , Sulfotransferases , Sulfotransferases/metabolism , Sulfotransferases/genetics , Heparin/metabolism , Heparin/biosynthesis , Anticoagulants/metabolism , Anticoagulants/chemistry , Escherichia coli/genetics , Escherichia coli/metabolism , Metabolic Engineering/methods , Humans , Polysaccharides/metabolism , Polysaccharides/biosynthesis , Polysaccharides/chemistry , Mutagenesis, Site-Directed , Protein Engineering/methods , Disaccharides/metabolism , Disaccharides/biosynthesis , Disaccharides/chemistry , Recombinant Proteins/metabolism , Recombinant Proteins/genetics
17.
Int J Mol Sci ; 25(9)2024 May 03.
Article En | MEDLINE | ID: mdl-38732217

The Euganean Thermal District, situated in North-East Italy, is one of Europe's largest and oldest thermal centres. The topical application of its therapeutic thermal muds is recognised by the Italian Health System as a beneficial treatment for patients suffering from arthro-rheumatic diseases. Polysaccharides produced by the mud microbiota have been recently identified as anti-inflammatory bioactive molecules. In this paper we analysed the efficacy of Microbial-Polysaccharides (M-PS) derived from mature muds obtained at different maturation temperatures, both within and outside the codified traditional mud maturation range. M-PSs were extracted from six mature muds produced by five spas of the Euganean Thermal District and investigated for their chemical properties, monosaccharide composition and in vivo anti-inflammatory potential, using the zebrafish model organism. Additionally, mature muds were characterized for their microbiota composition using Next-Generation Sequencing. The results showed that all M-PSs exhibit similar anti-inflammatory potential, referable to their comparable chemical composition. This consistency was observed despite changes in cyanobacteria populations, suggesting a possible role of the entire microbial community in shaping the properties of these biomolecules. These findings highlight the importance of scientific research in untangling the origins of the therapeutic efficacy of Euganean Thermal muds in the treatment of chronic inflammatory conditions.


Anti-Inflammatory Agents , Zebrafish , Animals , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/chemistry , Italy , Polysaccharides, Bacterial/pharmacology , Polysaccharides, Bacterial/chemistry , Microbiota/drug effects , Polysaccharides/pharmacology , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Mud Therapy
18.
ACS Nano ; 18(19): 12412-12426, 2024 May 14.
Article En | MEDLINE | ID: mdl-38693619

Glycans play vital roles in nearly all life processes of multicellular organisms, and understanding these activities is inseparable from elucidating the biological significance of glycans. However, glycan research has lagged behind that of DNA and protein due to the challenges posed by structural heterogeneity and isomerism (i.e., structures with equal molecular weights) the lack of high-efficiency structural analysis techniques. Nanopore technology has emerged as a sensitive single-molecule biosensor, shining a light on glycan analysis. However, a significant number of glycans are small and uncharged, making it challenging to elicit identifiable nanopore signals. Here we introduce a R-binaphthyl tag into glycans, which enhances the cation-π interaction between the derivatized glycan molecules and the nanopore interface, enabling the detection of neutral glycans with an aerolysin nanopore. This approach allows for the distinction of di-, tri-, and tetrasaccharides with monosaccharide resolution and has the potential for group discrimination, the monitoring of enzymatic transglycosylation reactions. Notably, the aerolysin mutant T240R achieves unambiguous identification of six disaccharide isomers, trisaccharide and tetrasaccharide linkage isomers. Molecular docking simulations reveal that multiple noncovalent interactions occur between residues R282, K238, and R240 and the glycans and R-binaphthyl tag, significantly slowing down their translocation across the nanopore. Importantly, we provide a demonstration of the kinetic translocation process of neutral glycan isomers, establishing a solid theoretical foundation for glycan nanopore analysis. The development of our technology could promote the analysis of glycan structural isomers and has the potential for nanopore-based glycan structural determination and sequencing.


Bacterial Toxins , Nanopores , Polysaccharides , Pore Forming Cytotoxic Proteins , Polysaccharides/chemistry , Bacterial Toxins/chemistry , Bacterial Toxins/genetics , Pore Forming Cytotoxic Proteins/chemistry , Pore Forming Cytotoxic Proteins/genetics , Molecular Docking Simulation , Mutation
19.
Molecules ; 29(9)2024 Apr 26.
Article En | MEDLINE | ID: mdl-38731484

In this study, we developed a green and multifunctional bioactive nanoemulsion (BBG-NEs) of Blumea balsamifera oil using Bletilla striata polysaccharide (BSP) and glycyrrhizic acid (GA) as natural emulsifiers. The process parameters were optimized using particle size, PDI, and zeta potential as evaluation parameters. The physicochemical properties, stability, transdermal properties, and bioactivities of the BBG-NEs under optimal operating conditions were investigated. Finally, network pharmacology and molecular docking were used to elucidate the potential molecular mechanism underlying its wound-healing properties. After parameter optimization, BBG-NEs exhibited excellent stability and demonstrated favorable in vitro transdermal properties. Furthermore, it displayed enhanced antioxidant and wound-healing effects. SD rats wound-healing experiments demonstrated improved scab formation and accelerated healing in the BBG-NE treatment relative to BBO and emulsifier groups. Pharmacological network analyses showed that AKT1, CXCL8, and EGFR may be key targets of BBG-NEs in wound repair. The results of a scratch assay and Western blotting assay also demonstrated that BBG-NEs could effectively promote cell migration and inhibit inflammatory responses. These results indicate the potential of the developed BBG-NEs for antioxidant and skin wound applications, expanding the utility of natural emulsifiers. Meanwhile, this study provided a preliminary explanation of the potential mechanism of BBG-NEs to promote wound healing through network pharmacology and molecular docking, which provided a basis for the mechanistic study of green multifunctional nanoemulsions.


Antioxidants , Emulsifying Agents , Emulsions , Glycyrrhizic Acid , Molecular Docking Simulation , Wound Healing , Wound Healing/drug effects , Animals , Emulsions/chemistry , Emulsifying Agents/chemistry , Emulsifying Agents/pharmacology , Rats , Antioxidants/pharmacology , Antioxidants/chemistry , Antioxidants/chemical synthesis , Glycyrrhizic Acid/pharmacology , Glycyrrhizic Acid/chemistry , Polysaccharides/chemistry , Polysaccharides/pharmacology , Green Chemistry Technology , Humans , Rats, Sprague-Dawley , Nanoparticles/chemistry , Plant Oils/chemistry , Plant Oils/pharmacology , Fabaceae/chemistry , Male , Particle Size , Cell Movement/drug effects
20.
Molecules ; 29(9)2024 Apr 27.
Article En | MEDLINE | ID: mdl-38731509

The aim of this study was to develop microcapsules containing juniper or black pepper essential oils, using a combination of faba bean protein and chia seed polysaccharides (in ratios of 1:1, 1:2, 2:1). By synergizing these two polymers, our goal was to enhance the efficiency of essential oil microencapsulation, opening up various applications in the food industry. Additionally, we aimed to investigate the influence of different polymer mixing ratios on the properties of the resulting microcapsules and the course of the complex coacervation process. To dissolve the essential oils and limit their evaporation, soybean and rapeseed oils were used. The powders resulting from the freeze-drying of coacervates underwent testing to assess microencapsulation efficiency (65.64-87.85%), density, flowability, water content, solubility, and hygroscopicity. Additionally, FT-IR and DSC analyses were conducted. FT-IR analysis confirmed the interactions between the components of the microcapsules, and these interactions were reflected in their high thermal resistance, especially at a protein-to-polysaccharide ratio of 2:1 (177.2 °C). The water content in the obtained powders was low (3.72-7.65%), but it contributed to their hygroscopicity (40.40-76.98%).


Capsules , Drug Compounding , Oils, Volatile , Plant Proteins , Polysaccharides , Salvia , Seeds , Vicia faba , Polysaccharides/chemistry , Seeds/chemistry , Vicia faba/chemistry , Drug Compounding/methods , Oils, Volatile/chemistry , Plant Proteins/chemistry , Salvia/chemistry , Capsules/chemistry , Solubility , Spectroscopy, Fourier Transform Infrared , Water/chemistry
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