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1.
Molecules ; 29(9)2024 May 02.
Article in English | MEDLINE | ID: mdl-38731598

ABSTRACT

Obtaining high-added value compounds from agricultural waste receives increasing attention, as it can both improve resource utilization efficiency and reduce waste generation. In this study, polysaccharides are extracted from the discarded roots of Abelmoschus manihot (L.) by the high-efficiency ultrasound-assisted extraction (UAE). The optimized condition was determined as solid-liquid ratio SL ratio = 1:20, temperature T = 30 °C and time T = 40 min, achieving an extraction yield of 13.41%. Composition analysis revealed that glucose (Glc, 44.65%), rhamnose (Rha, 26.30%), galacturonic acid (GalA, 12.50%) and galactose (Gal, 9.86%) are the major monosaccharides of the extract. The extract showed a low degree of esterification (DE) value of 40.95%, and its Fourier-transform infrared (FT-IR) spectrum exhibited several characteristic peaks of polysaccharides. Inspired by the wide cosmetic applications of polysaccharides, the skincare effect of the extract was evaluated via the moisture retention, total phenolic content (TPC) quantification, 2,2-Diphenyl-1-picrylhydrazyl (DPPH)-free radical scavenging activity, anti-hyaluronidase and anti-elastase activity experiments. The extract solutions demonstrated a 48 h moisture retention rate of 10.75%, which is superior to that of commercially available moisturizer hyaluronic acid (HA). Moreover, both the TPC value of 16.16 mg GAE/g (dw) and DPPH-free radical scavenging activity of 89.20% at the concentration of 2 mg/mL indicated the strong anti-oxidant properties of the extract. Furthermore, the anti-hyaluronidase activity and moderate anti-elastase activity were determined as 72.16% and 42.02%, respectively. In general, in vitro skincare effect experiments suggest moisturizing, anti-oxidant, anti-radical and anti-aging activities of the A. manihot root extract, indicating its potential applications in the cosmetic industry.


Subject(s)
Abelmoschus , Antioxidants , Plant Extracts , Plant Roots , Polysaccharides , Polysaccharides/chemistry , Polysaccharides/pharmacology , Polysaccharides/isolation & purification , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Roots/chemistry , Abelmoschus/chemistry , Antioxidants/chemistry , Antioxidants/pharmacology , Spectroscopy, Fourier Transform Infrared , Skin Care/methods , Rhamnose/chemistry , Galactose , Hexuronic Acids/chemistry , Phenols/chemistry , Phenols/analysis , Phenols/pharmacology , Humans
2.
Exp Dermatol ; 33(5): e15098, 2024 May.
Article in English | MEDLINE | ID: mdl-38770557

ABSTRACT

Healing of complex wounds requires dressings that must, at least, not hinder and should ideally promote the activity of key healing cells, in particular fibroblasts. This in vitro study assessed the effects of three wound-dressings (a pure Ca2+ alginate: Algostéril®, a Ca2+ alginate + carboxymethylcellulose: Biatain alginate® and a polyacrylate impregnated with lipido-colloid matrix: UrgoClean®) on dermal fibroblast activity. The results showed the pure calcium alginate to be non-cytotoxic, whereas the other wound-dressings showed moderate to strong cytotoxicity. The two alginates stimulated fibroblast migration and proliferation, whereas the polyacrylate altered migration and had no effect on proliferation. The pure Ca2+ alginate significantly increased the TGF-ß-induced fibroblast activation, which is essential to healing. This activation was confirmed by a significant increase in Vascular endothelial growth factor (VEGF) secretion and a higher collagen production. The other dressings reduced these fibroblast activities. The pure Ca2+ alginate was also able to counteract the inhibitory effect of NK cell supernatants on fibroblast migration. These in vitro results demonstrate that tested wound-dressings are not equivalent for fibroblast activation. Only Algostéril was found to promote all the fibroblast activities tested, which could contribute to its healing efficacy demonstrated in the clinic.


Subject(s)
Alginates , Cell Movement , Cell Proliferation , Fibroblasts , Vascular Endothelial Growth Factor A , Wound Healing , Fibroblasts/drug effects , Wound Healing/drug effects , Humans , Alginates/pharmacology , Cell Movement/drug effects , Cell Proliferation/drug effects , Vascular Endothelial Growth Factor A/metabolism , Collagen/metabolism , Bandages , Transforming Growth Factor beta/metabolism , Carboxymethylcellulose Sodium , Cells, Cultured , Killer Cells, Natural/drug effects , Acrylic Resins , Hexuronic Acids , Glucuronic Acid , Skin
3.
Org Biomol Chem ; 22(19): 3986-3994, 2024 05 15.
Article in English | MEDLINE | ID: mdl-38695061

ABSTRACT

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.


Subject(s)
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
4.
Plant Physiol Biochem ; 211: 108694, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38714131

ABSTRACT

Using natural clinoptilolite (NCP) as a carrier and alginate (Alg)-calcium as an active species, the porous silicon calcium alginate nanocomposite (Alg-Ca-NCP) was successfully fabricated via adsorption-covalence-hydrogen bond. Its structural features and physicochemical properties were detailed investigated by various characterizations. The results indicated that Alg-Ca-NCP presented the disordered lamellar structures with approximately uniform particles in size of 300-500 nm. Specially, their surface fractal evolutions between the irregular roughness and dense structures were demonstrated via the SAXS patterns. The results elucidated that the abundant micropores of NCP were beneficial for unrestricted diffusing of Alg-Ca, which was conducive to facilitate a higher loading and sustainable releasing. The Ca content of leaf mustard treated with Alg-Ca-NCP-0.5 was 484.5 mg/100g on the 21st day, higher than that by water (CK) and CaCl2 solution treatments, respectively. Meanwhile, the prepared Alg-Ca-NCPs presented the obvious anti-aging effects on peroxidase drought stress of mustard leaves. These demonstrations provided a simple and effective method to synthesize Alg-Ca-NCPs as delivery nanocomposites, which is useful to improve the weak absorption and low utilization of calcium alginate by plants.


Subject(s)
Alginates , Mustard Plant , Zeolites , Alginates/chemistry , Alginates/pharmacology , Zeolites/chemistry , Zeolites/pharmacology , Mustard Plant/metabolism , Mustard Plant/drug effects , Mustard Plant/chemistry , Plant Leaves/metabolism , Plant Leaves/drug effects , Plant Leaves/chemistry , Porosity , Brassica/metabolism , Brassica/drug effects , Brassica/growth & development , Glucuronic Acid/chemistry , Nanocomposites/chemistry , X-Ray Diffraction , Hexuronic Acids/chemistry , Hexuronic Acids/metabolism
5.
J Biomed Mater Res B Appl Biomater ; 112(6): e35433, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38817048

ABSTRACT

Ex vivo tissue engineering is an effective therapeutic approach for the treatment of severe cartilage diseases that require tissue replenishment or replacement. This strategy demands scaffolds that are durable enough for long-term cell culture to form artificial tissue. Additionally, such scaffolds must be biocompatible to prevent the transplanted matrix from taking a toll on the patient's body. From the viewpoint of structure and bio-absorbability, a ß-tricalcium phosphate (ß-TCP) fiber scaffold (ßTFS) is expected to serve as a good scaffold for tissue engineering. However, the fragility and high solubility of ß-TCP fibers make this matrix unsuitable for long-term cell culture. To solve this problem, we developed an alginate-coated ß-TCP fiber scaffold (ßTFS-Alg). To assess cell proliferation and differentiation in the presence of ßTFS-Alg, we characterized ATDC5 cells, a chondrocyte-like cell line, when grown in this matrix. We found that alginate coated the surface of ßTFS fiber and suppressed the elution of Ca2+ from ß-TCP fibers. Due to the decreased solubility of ßTFS-Alg compared with ß-TCP, the former provided an improved scaffold for long-term cell culture. Additionally, we observed superior cell proliferation and upregulation of chondrogenesis marker genes in ATDC5 cells cultured in ßTFS-Alg. These results suggest that ßTFS-Alg is suitable for application in tissue culture.


Subject(s)
Alginates , Calcium Phosphates , Tissue Scaffolds , Calcium Phosphates/chemistry , Calcium Phosphates/pharmacology , Alginates/chemistry , Tissue Scaffolds/chemistry , Cell Proliferation , Mice , Glucuronic Acid/chemistry , Animals , Hexuronic Acids/chemistry , Coated Materials, Biocompatible/chemistry , Coated Materials, Biocompatible/pharmacology , Cell Line , Chondrocytes/cytology , Chondrocytes/metabolism , Tissue Engineering , Materials Testing , Cell Differentiation , Humans , Cell Culture Techniques
6.
Food Chem ; 452: 139282, 2024 Sep 15.
Article in English | MEDLINE | ID: mdl-38723562

ABSTRACT

Acrylamide (AA) is a neoformed compound in heated foods, mainly produced between asparagine (Asn) and glucose (Glc) during the Maillard reaction. Galacturonic acid (GalA), the major component of pectin, exhibits high activity in AA formation. This study investigated the pathway for AA formation between GalA and Asn. Three possible pathways were proposed: 1) The carbonyl group of GalA directly interacts with Asn to produce AA; 2) GalA undergoes an oxidative cleavage reaction to release α-dicarbonyl compounds, which subsequently leads to AA production; 3) 5-formyl-2-furancarboxylic acid, the thermal degradation product of GalA, reacts with Asn to generate AA. Structural analysis revealed that the COOH group in GalA accelerated intramolecular protonation and electron transfer processes, thereby increasing the formation of AA precursors such as decarboxylated Schiff base and α-dicarbonyl compounds, promoting AA formation. This study provides a theoretical basis and new insights into the formation and control of AA.


Subject(s)
Acrylamide , Hexuronic Acids , Acrylamide/chemistry , Hexuronic Acids/chemistry , Maillard Reaction , Asparagine/chemistry , Hot Temperature , Pectins/chemistry , Molecular Structure
7.
Int J Biol Macromol ; 269(Pt 2): 131890, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38692534

ABSTRACT

The rheological and morphological characteristics of Ca-crosslinked alginate hydrogels with two different M/G ratios, α-L-guluronate (G)-rich and ß-D-mannuronate (M)-rich, each with one alginic acid concentration, were investigated. It was found that the stiffness and elasticity of alginate hydrogels are derived from the thickness and density of the fibril network structures. In aqueous alginate solution, ball-like aggregates of alginates are present. Time-resolved small-angle X-ray scattering and time-domain nuclear magnetic resonance measurements suggest that the disaggregation of alginate aggregates and loose fibrillation occur in the early stage of the sol-gel transition. After these induction stage, direct gelation is finally caused by the formation of the egg-box junction. G-rich alginate hydrogel has a higher stiffness and a thicker and denser fibril network structure than M-rich alginate hydrogel. The former also exhibits faster and more significant changes in physical properties during the sol-gel transition.


Subject(s)
Alginates , Hydrogels , Phase Transition , Rheology , Alginates/chemistry , Hydrogels/chemistry , Scattering, Small Angle , Hexuronic Acids
8.
Int J Biol Macromol ; 269(Pt 2): 131918, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38697418

ABSTRACT

Polygalacturonases (PGs) can modulate chemistry and mechanical properties of the plant cell wall through the degradation of pectins, one of its major constituents. PGs are largely used in food, beverage, textile, and paper industries to increase processes' performances. To improve the use of PGs, knowledge of their biochemical, structural and functional features is of prime importance. Our study aims at characterizing SmoPG1, a polygalacturonase from Selaginella moellendorffii, that belongs to the lycophytes. Transcription data showed that SmoPG1 was mainly expressed in S. moellendorffii shoots while phylogenetic analyses suggested that SmoPG1 is an exo-PG, which was confirmed by the biochemical characterization following its expression in heterologous system. Indeed, LC-MS/MS oligoprofiling using various pectic substrates identified galacturonic acid (GalA) as the main hydrolysis product. We found that SmoPG1 was most active on polygalacturonic acid (PGA) at pH 5, and that its activity could be modulated by different cations (Ca2+, Cu2+, Fe2+, Mg2+, Mn2+, Na2+, Zn2+). In addition, SmoPG1 was inhibited by green tea catechins, including (-)-epigallocatechin-3-gallate (EGCG). Docking analyses and MD simulations showed in detail amino acids responsible for the SmoPG1-EGCG interaction. Considering its expression yield and activity, SmoPG1 appears as a prime candidate for the industrial production of GalA.


Subject(s)
Pectins , Polygalacturonase , Selaginellaceae , Polygalacturonase/metabolism , Polygalacturonase/chemistry , Polygalacturonase/genetics , Selaginellaceae/chemistry , Selaginellaceae/genetics , Selaginellaceae/enzymology , Pectins/metabolism , Pectins/chemistry , Phylogeny , Substrate Specificity , Molecular Docking Simulation , Amino Acid Sequence , Hydrogen-Ion Concentration , Hydrolysis , Hexuronic Acids
9.
Braz Dent J ; 35: e245461, 2024.
Article in English | MEDLINE | ID: mdl-38775590

ABSTRACT

This study aimed to evaluate the osteogenic potential of hydroxyapatite (HA), Alginate (Alg), and Gelatine (Gel) composite in a critical-size defect model in rats. Twenty-four male rats were divided into three groups: a negative control with no treatment (Control group), a positive control treated with deproteinized bovine bone mineral (DBBM group), and the experimental group treated with the new HA-Alg-Gel composite (HA-Alg-Gel group). A critical size defect (8.5mm) was made in the rat's calvaria, and the bone formation was evaluated by in vivo microcomputed tomography analysis (µCT) after 1, 15, 45, and 90 days. After 90 days, the animals were euthanized and histological and histomorphometric analyses were performed. A higher proportion of mineralized tissue/biomaterial was observed in the DBBM group when compared to the HA-Alg-Gel and Control groups in the µCT analysis during all analysis periods. However, no differences were observed in the mineralized tissue/biomaterial proportion observed on day 1 (immediate postoperative) in comparison to later periods of analysis in all groups. In the histomorphometric analysis, the HA-Alg-Gel and Control groups showed higher bone formation than the DBBM group. Moreover, in histological analysis, five samples of the HA-Alg-Gal group exhibited formed bone spicules adjacent to the graft granules against only two of eight samples in the DBBM group. Both graft materials ensured the maintenance of defect bone thickness, while a tissue thickness reduction was observed in the control group. In conclusion, this study demonstrated the osteoconductive potential of HA-Alg-Gel bone graft by supporting new bone formation around its particles.


Subject(s)
Alginates , Bone Regeneration , Durapatite , Gelatin , Skull , X-Ray Microtomography , Animals , Bone Regeneration/drug effects , Durapatite/pharmacology , Skull/surgery , Skull/diagnostic imaging , Rats , Male , Biocompatible Materials , Glucuronic Acid , Rats, Wistar , Hexuronic Acids , Osteogenesis/drug effects , Bone Substitutes
10.
Tissue Eng Part C Methods ; 30(5): 229-237, 2024 May.
Article in English | MEDLINE | ID: mdl-38568845

ABSTRACT

Modeling organ-blood barriers through the inclusion of microvessel networks within in vitro tissue models could lead to more physiologically accurate results, especially since organ-blood barriers are crucial to the normal function, drug transport, and disease states of vascularized organs. Microvessel networks are difficult to form, since they push the practical limits of most fabrication methods, and it is difficult to coax vascular cells to self-assemble into structures larger than capillaries. Here, we present a method for rapidly forming networks of microvessel-like structures using sacrificial alginate structures. Specifically, we encapsulated endothelial cells within short alginate threads, and then embedded them in collagen gel. Following enzymatic degradation of the alginate, the collagen gel contained a network of hollow channels seeded with cells, all surrounding a perfusable central channel. This method uses a 3D-printed coaxial extruder and syringe pumps to generate short threads in a way that is repeatable and easily transferrable to other labs. The cell-laden, sacrificial alginate threads can be frozen after fabrication and thawed before embedding without significant loss of cell viability. The ability to freeze the threads enables future scale-up and ease of use. Within millifluidic devices that restrict access to media, the threads enhance cell survival under static conditions. These results indicate the potential for use of this method in a range of tissue engineering applications.


Subject(s)
Alginates , Microvessels , Tissue Engineering , Alginates/chemistry , Microvessels/cytology , Humans , Tissue Engineering/methods , Human Umbilical Vein Endothelial Cells/cytology , Human Umbilical Vein Endothelial Cells/metabolism , Tissue Scaffolds/chemistry , Endothelial Cells/cytology , Endothelial Cells/metabolism , Glucuronic Acid/chemistry , Hexuronic Acids/chemistry , Cell Survival , Animals , Collagen/chemistry
11.
J Gen Virol ; 105(4)2024 Apr.
Article in English | MEDLINE | ID: mdl-38656455

ABSTRACT

Porcine epidemic diarrhea (PED) is a serious disease in piglets that leads to high mortality. An effective measure that provides higher IgA levels in the intestine and milk is required to decrease losses. Porcine epidemic diarrhea virus (PEDV) was dissolved in calcium alginate (Alg) and combined with chitosan (CS) via electrostatic interactions between cationic chitosan and anionic alginate to create a porous gel (Alg-CS+PEDV). The gel was used to immunize mice orally or in combination with subcutaneous injections of inactivated PEDV vaccine. At 12 and 24 days after immunization, levels of IgA and IgG in Alg-CS+PEDV were higher than with normal PEDV oral administration. At 24 days after immunization, the concentration of IFN-γ in Alg-CS+PEDV was higher than with normal PEDV oral administration. Furthermore, oral administration combining subcutaneous immunization induced higher levels of IgG and IgA than oral administration alone. Our study provides a new method for the preparation and administration of oral vaccines to achieve enhanced mucosal immunity against PEDV.


Subject(s)
Alginates , Antibodies, Viral , Chitosan , Immunity, Mucosal , Immunoglobulin A , Immunoglobulin G , Porcine epidemic diarrhea virus , Viral Vaccines , Animals , Administration, Oral , Porcine epidemic diarrhea virus/immunology , Alginates/administration & dosage , Chitosan/administration & dosage , Mice , Viral Vaccines/immunology , Viral Vaccines/administration & dosage , Antibodies, Viral/immunology , Immunoglobulin A/immunology , Immunoglobulin G/blood , Swine , Coronavirus Infections/immunology , Coronavirus Infections/prevention & control , Coronavirus Infections/veterinary , Coronavirus Infections/virology , Vaccines, Inactivated/administration & dosage , Vaccines, Inactivated/immunology , Swine Diseases/immunology , Swine Diseases/prevention & control , Swine Diseases/virology , Female , Gels/administration & dosage , Mice, Inbred BALB C , Interferon-gamma/immunology , Glucuronic Acid/administration & dosage , Hexuronic Acids/administration & dosage
12.
Biomed Mater ; 19(3)2024 Apr 26.
Article in English | MEDLINE | ID: mdl-38574669

ABSTRACT

Recently,in vitromodels of intestinal mucosa have become important tools for drug screening and studying the physiology and pathology of the intestine. These models enable the examination of cellular behavior in diseased states or in reaction to alterations in the microenvironment, potentially serving as alternatives to animal models. One of the major challenges in constructing physiologically relevantin vitromodels of intestinal mucosa is the creation of three-dimensional microstructures that accurately mimic the integration of intestinal epithelium and vascularized stroma. Here, core-shell alginate (Alg) microspheres were generated to create the compartmentalized extracellular matrix microenvironment needed to simulate the epithelial and vascularized stromal compartments of the intestinal mucosa. We demonstrated that NIH-3T3 and human umbilical vein endothelial cells embedded in the core of the microspheres can proliferate and develop a vascular network, while human colorectal adenocarcinoma cells (Caco-2) can form an epithelial monolayer in the shell. Compared to Caco-2 monolayer encapsulated within the shell, the presence of the vascularized stroma enhances their proliferation and functionality. As such, our core-shell Alg microspheres provide a valuable method for generatingin vitromodels of vascularized intestinal mucosa with epithelial and vascularized stroma arranged in a spatially relevant manner and demonstrating near-physiological functionality.


Subject(s)
Alginates , Cell Proliferation , Human Umbilical Vein Endothelial Cells , Intestinal Mucosa , Microspheres , Tissue Engineering , Alginates/chemistry , Humans , Intestinal Mucosa/metabolism , Animals , Mice , Caco-2 Cells , Tissue Engineering/methods , NIH 3T3 Cells , Extracellular Matrix/metabolism , Tissue Scaffolds/chemistry , Hexuronic Acids/chemistry
13.
Molecules ; 29(7)2024 Apr 01.
Article in English | MEDLINE | ID: mdl-38611855

ABSTRACT

Quinoa, known as the "golden grain" for its high nutritional value, has polysaccharides as one of its sources of important nutrients. However, the biological functions of quinoa polysaccharides remain understudied. In this study, two crude polysaccharide extracts of quinoa (Q-40 and Q-60) were obtained through sequential precipitation with 40% and 60% ethanol, with purities of 58.29% (HPLC) and 62.15% (HPLC) and a protein content of 8.27% and 9.60%, respectively. Monosaccharide analysis revealed that Q-40 contained glucose (Glc), galacturonic acid (GalA), and arabinose (Ara) in a molar ratio of 0.967:0.027:0.006. Q-60 was composed of xylose (xyl), arabinose (Ara), galactose, and galacturonic acid (GalA) with a molar ratio of 0.889:0.036:0.034:0.020. The average molecular weight of Q-40 ranged from 47,484 to 626,488 Da, while Q-60 showed a range of 10,025 to 47,990 Da. Rheological experiments showed that Q-40 exhibited higher viscosity, while Q-60 demonstrated more elastic properties. Remarkably, Q-60 showed potent antioxidant abilities, with scavenging rates of 98.49% for DPPH and 57.5% for ABTS. Antibacterial experiments using the microdilution method revealed that Q-40 inhibited the growth of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli), while Q-60 specifically inhibited MRSA. At lower concentrations, both polysaccharides inhibited MDA (MD Anderson Cancer Center) cell proliferation, but at higher concentrations, they promoted proliferation. Similar proliferation-promoting effects were observed in HepG2 cells. The research provides important information in the application of quinoa in the food and functional food industries.


Subject(s)
Chenopodium quinoa , Hexuronic Acids , Methicillin-Resistant Staphylococcus aureus , Arabinose , Escherichia coli , Edible Grain
14.
Sci Rep ; 14(1): 9182, 2024 04 22.
Article in English | MEDLINE | ID: mdl-38649422

ABSTRACT

In order to obtain high yield pomelo peel pectin with better physicochemical properties, four pectin extraction methods, including hot acid extraction (HAE), microwave-assisted extraction (MAE), ultrasound-assisted extraction, and enzymatic assisted extraction (EAE) were compared. MAE led to the highest pectin yield (20.43%), and the lowest pectin recovery was found for EAE (11.94%). The physicochemical properties of pomelo peel pectin obtained by different methods were also significantly different. Pectin samples obtained by MAE had the highest methoxyl content (8.35%), galacturonic acid content (71.36%), and showed a higher apparent viscosity, thermal and emulsion stability. The pectin extracted by EAE showed the highest total phenolic content (12.86%) and lowest particle size (843.69 nm), showing higher DPPH and ABTS scavenging activities than other extract methods. The pectin extracted by HAE had the highest particle size (966.12 nm) and degree of esterification (55.67%). However, Fourier-transform infrared spectroscopy showed that no significant difference occurred among the different methods in the chemical structure of the extracted pectin. This study provides a theoretical basis for the industrial production of pomelo peel pectin.


Subject(s)
Citrus , Hexuronic Acids , Pectins , Pectins/chemistry , Pectins/isolation & purification , Citrus/chemistry , Viscosity , Particle Size , Microwaves , Spectroscopy, Fourier Transform Infrared , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Chemical Fractionation/methods , Chemical Phenomena , Fruit/chemistry , Antioxidants/chemistry , Antioxidants/isolation & purification , Antioxidants/pharmacology , Phenols/analysis , Phenols/chemistry , Phenols/isolation & purification , Esterification
15.
J Biosci Bioeng ; 137(6): 463-470, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38570220

ABSTRACT

The choice of sterilization method for hydrogels used for cell culture influences the ease of preparing the gel. We prepared interpenetrating gelatin/calcium alginate hydrogels containing 1% (w/v) alginate and 1-16% (w/v) gelatin by molding with the mixture of gelatin/sodium alginate solution, followed by the addition of calcium ions by incubation in calcium chloride solution. It is the simplest method to prepare autoclavable gelatin/sodium hydrogel. We measured various properties of the hydrogels including volume, Young's modulus in the compression test, storage modulus, and loss modulus in the dynamic viscoelasticity measurement. The gelatin/alginate hydrogel can be easily fabricated into any shape by this method. After autoclave treatment, the hydrogel was shrunk to smaller than the original shape in similar figures. The shape of the gelatin/alginate hydrogel can be designed into any shape with the reduction ratio of the volume. Human osteosarcoma (HOS) cells adhered to the gelatin/alginate hydrogel and then proliferated. Gelatin/calcium alginate hydrogels with a high concentration are considered to be autoclavable culture substrates because of their low deformation and gelatin elution rate after autoclaving and the high amount of cells attached to the hydrogels.


Subject(s)
Alginates , Gelatin , Hydrogels , Tissue Scaffolds , Gelatin/chemistry , Alginates/chemistry , Hydrogels/chemistry , Humans , Tissue Scaffolds/chemistry , Cell Line, Tumor , Sterilization , Cell Proliferation/drug effects , Glucuronic Acid/chemistry , Tissue Engineering/methods , Hexuronic Acids/chemistry , Elastic Modulus , Cell Adhesion
16.
J Mater Sci Mater Med ; 35(1): 19, 2024 Mar 25.
Article in English | MEDLINE | ID: mdl-38526655

ABSTRACT

The efficacy of stem-cell therapy depends on the ability of the transplanted cells to escape early immunological reactions and to be retained at the site of transplantation. The use of tissue engineering scaffolds or injectable biomaterials as carriers has been proposed, but they still present limitations linked to a reliable manufacturing process, surgical practice and clinical outcomes. Alginate microbeads are potential candidates for the encapsulation of mesenchymal stromal cells with the aim of providing a delivery carrier suitable for minimally-invasive and scaffold-free transplantation, tissue-adhesive properties and protection from the immune response. However, the formation of stable microbeads relies on the cross-linking of alginate with divalent calcium ions at concentrations that are toxic for the cells, making control over the beads' size and a single-cell encapsulation unreliable. The present work demonstrates the efficiency of an innovative, high throughput, and reproducible microfluidic system to produce single-cell, calcium-free alginate coatings of human mesenchymal stromal cells. Among the various conditions tested, visible light and confocal microscopy following staining of the cell nuclei by DAPI showed that the microfluidic system yielded an optimal single-cell encapsulation of 2000 cells/min in 2% w/v alginate microcapsules of reproducible morphology and an average size of 28.2 ± 3.7 µm. The adhesive properties of the alginate microcapsules, the viability of the encapsulated cells and their ability to escape the alginate microcapsule were demonstrated by the relatively rapid adherence of the beads onto tissue culture plastic and the cells' ability to gradually disrupt the microcapsule shell after 24 h and proliferate. To mimic the early inflammatory response upon transplantation, the encapsulated cells were exposed to proliferating macrophages at different cell seeding densities for up to 2 days and the protection effect of the microcapsule on the cells assessed by time-lapse microscopy showing a shielding effect for up to 48 h. This work underscores the potential of microfluidic systems to precisely encapsulate cells by good manufacturing practice standards while favouring cell retention on substrates, viability and proliferation upon transplantation.


Subject(s)
Mesenchymal Stem Cells , Microfluidics , Humans , Cell Encapsulation , Capsules , Bone Marrow , Alginates/chemistry , Hexuronic Acids/chemistry , Cell Survival , Glucuronic Acid/chemistry
17.
Plant Physiol Biochem ; 208: 108495, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38452451

ABSTRACT

Solanum lycopersicum (Tomato) leaves and stems are considered waste. Valorization of this waste can be achieved by for example the extraction of proteins. This prospect is promising but currently not feasible, since protein extraction yields from tomato leaves are low, amongst other due to the (physical) barrier formed by the plant cell walls. However, the molecular aspects of the relationship between cell wall properties and protein extractability from tomato leaves are currently not clear and thus objective of this study. To fill this knowledge gap the biochemical composition of plant cell walls was measured and related to protein extraction yields at different plant ages, leaf positions, and across different tomato accessions, including two Solanum lycopersicum cultivars and the wildtype species S. pimpinellifolium and S. pennellii. For all genotypes, protein extraction yields from tomato leaves were the highest in young tissues, with a decreasing trend towards older plant material. This decrease of protein extraction yield was accompanied by a significant increase of arabinose and galacturonic acid content and a decrease of galactose content in the cell walls of old-vs-young tissues. This resulted in strong negative correlations between protein extraction yield and the content of arabinose and galacturonic acid in the cell wall, and a positive correlation between the content of galactose and protein extraction yield. Overall, these results point to the importance of the pectin network on protein extractability, making pectin a potential breeding target for enhancing protein extractability from tomato leaves.


Subject(s)
Hexuronic Acids , Solanum lycopersicum , Solanum lycopersicum/genetics , Arabinose , Galactose , Plant Breeding , Cell Wall/metabolism , Plant Leaves/metabolism , Pectins/metabolism
18.
Int J Biol Macromol ; 266(Pt 1): 131164, 2024 May.
Article in English | MEDLINE | ID: mdl-38547940

ABSTRACT

The biological potency of pectin is intricately intertwined with its intricate molecular architecture. The fine structure of pectin is influenced by the extraction method, while the specific impact of these methods on the fine structure and the affected attributes thereof remains enigmatic. This study delves into the profound analysis of eight distinct extraction methods influence on the structure and biological activity of citrus peel pectin. The findings demonstrate that citric acid ultrasound-assisted microwave extraction yields pectin (PectinCA-US/MV) with higher viscosity and a dense, rigid chain. Pectin extracted with acetic acid ultrasound (PectinAA-US) and citric acid ultrasound (PectinCA-US) exhibits elevated galacturonic acid (GalA) levels and reduced D-galactose (Gal) content, enhancing antioxidant activity. Eight pectin-chitosan (CS) hydrogels, especially PectinCA-US/MV-CS, demonstrate commendable thermal stability, rheological properties, self-healing capability, and swelling behavior. This study characterizes citrus peel pectin properties from different extraction methods, laying a foundation for its application in food, pharmaceuticals, and industry.


Subject(s)
Antioxidants , Citrus , Hexuronic Acids , Microwaves , Pectins , Pectins/chemistry , Pectins/isolation & purification , Pectins/pharmacology , Antioxidants/chemistry , Antioxidants/pharmacology , Citrus/chemistry , Viscosity , Hydrogels/chemistry , Citric Acid/chemistry , Chitosan/chemistry , Rheology , Ultrasonic Waves
19.
Int J Biol Macromol ; 262(Pt 1): 130006, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38331067

ABSTRACT

The processing characteristics of yogurt are closely related to the composition and arrangement of exopolysaccharides (EPS) in lactic acid bacteria (LAB). To fully understand and develop the functional properties of EPS and to study the effect of EPS molecular weight on yogurt and its mechanism, the physicochemical properties of high molecular weight EPS-LH43, medium molecular weight EPS-LH13, and low molecular weight EPS-LH23, as well as the gel properties and protein conformation of yogurt, were determined and analyzed in this experiment. The results indicate that EPS-LH43 and EPS-LH13 are both composed of mannose, rhamnose, galacturonic acid, glucose, and galactose. EPS-LH23 is composed of mannose, galacturonic acid, glucose, and galactose. Their Number-average Molecular Weight is 5.21 × 106 Da, 2.39 × 106 Da and 3.76 × 105 Da, respectively. In addition, all three types of EPS have good thermal stability and can improve the stability of casein. In addition, the analysis of the texture, particle size, potential, water holding capacity, rheology, low field nuclear magnetic resonance, microstructure, and flavor characteristics of yogurt confirmed the relationship between the molecular weight of LAB EPS and the gel properties of yogurt. Fluorescence spectrophotometer and circular dichroism analysis indicate that the different molecular weights of LAB EPS have different effects on protein structure, which is an intrinsic factor leading to significant differences in the gel properties of the three types of fermented milk. These findings provide new references for enhancing the understanding of the structure-activity relationship of EPS and indicate that EPS-LH43 can be used to improve the gel properties of dairy products.


Subject(s)
Hexuronic Acids , Lactobacillus helveticus , Yogurt , Yogurt/microbiology , Polysaccharides, Bacterial/chemistry , Molecular Weight , Galactose/analysis , Mannose , Glucose/analysis , Fermentation
20.
Int J Biol Macromol ; 263(Pt 2): 130452, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38417755

ABSTRACT

As a traditional Chinese medicinal and edible homologous plant, Onosma glomeratum Y. L. Liu has been used for treating lung diseases in Tibet. In this study, a pectin polysaccharide, OGY-LLPA, with a molecular weight of 62,184 Da, was isolated and characterized by GC-MS and NMR analysis. It mainly consists of galacturonic acid (GalA), galactose (Gal), rhamnose (Rha), and arabinose (Ara), with a linear main chain of galacturonic acid (homogalacturonan, HG) inserted by part of rhamnose galacturonic acid (rhamnogalacturonan, RG), attaching with arabinogalactan (AG) branches at RG-I. Both in the LPS-induced A549 cell model and LPS-induced pneumonia mouse model, OGY-LLPA demonstrated strong anti-inflammatory effects, even comparable to DEX, indicating its potential as an anti-pneumonia candidate agent. Moreover, low-dose OGY-LLPA alleviated LPS-induced pulmonary inflammation by inhibiting the NF-κB signaling pathway. Overall, these findings could not only contribute to the utilization of Onosma glomeratum Y. L. Liu., but also provides a theoretical basis for the treatment of inflammation-related diseases.


Subject(s)
Hexuronic Acids , NF-kappa B , Pneumonia , Mice , Animals , Lipopolysaccharides , Rhamnose , Polysaccharides/pharmacology , Polysaccharides/therapeutic use , Polysaccharides/analysis , Signal Transduction , Pneumonia/chemically induced , Pneumonia/drug therapy
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