Repair Effect of Umbilical Cord Mesenchymal Stem Cells Embedded in Hydrogel on Mouse Insulinoma 6 Cells Injured by Streptozotocin.

Umbilical cord mesenchymal stem cells (UC-MSCs) possess the capabilities of differentiation and immune modulation, which endow them with therapeutic potential in the treatment of type 2 diabetes mellitus (T2DM). In this study, to investigate the repair mechanism of UC-MSCs in hydrogel on pancreatic ß-cells in diabetes, mouse insulinoma 6 (MIN-6) cells damaged by streptozotocin (STZ) in vitro were used in co-culture with UC-MSCs in hydrogel (UC-MSCs + hydrogel). It was found that UC-MSCs + hydrogel had a significant repair effect on injured MIN-6 cells, which was better than the use of UC-MSCs alone (without hydrogel). After repair, the expression of superoxide dismutase (SOD) and catalase (CAT) as well as the total antioxidant capacity (T-AOC) of the repaired MIN-6 cells were increased, effectively reducing the oxidative stress caused by STZ. In addition, UC-MSCs + hydrogel were able to curb the inflammatory response by promoting the expression of anti-inflammatory factor IL-10 and reducing inflammatory factor IL-1ß. In addition, the expression of both nuclear antigen Ki67 for cell proliferation and insulin-related genes such as Pdx1 and MafA was increased in the repaired MIN-6 cells by UC-MSCs + hydrogel, suggesting that the repair effect promotes the proliferation of the injured MIN-6 cells. Compared with the use of UC-MSCs alone, UC-MSCs + hydrogel exhibit superior antioxidant stress resistance against injured MIN-6 cells, better proliferation effects and a longer survival time of UC-MSCs because the porous structure and hydrophilic properties of the hydrogel could affect the growth of cells and slow down their metabolic activities, resulting in a better repair effect on the injured MIN-6 cells.

A carboxymethyl chitosan/oxidized hyaluronic acid composite hydrogel dressing loading with stem cell exosome for chronic inflammation wounds healing.

Stem cell exosomes (Exo) play an important role in the transformation of macrophages, but the rapid clearance of Exo in vivo limits their therapeutic effects for chronic inflammation wounds healing. Here, stem cell Exo was isolated and introduced to a composite hydrogel including carboxymethyl chitosan (CMCS) and oxidized hyaluronic acid (OHA) through chemical cross-linking, which formed an Exo-loaded (CMCS/OHA/Exo) hydrogel. The CMCS/OHA/Exo hydrogel exhibited a function of Exo sustained release and an Exo protection within 6 days. This CMCS/OHA/Exo hydrogel was much better than CMCS/OHA hydrogel or Exo solution in macrophage cell phagocytosis, proliferation and migration in vitro, especially, played an obviously positive role in the transformation of macrophages compared with the reference groups. For the treatment of the chronic inflammation wounds in vivo, the CMCS/OHA/Exo hydrogel had the best results at wound heal rate and inhibiting the secretion of inflammatory factors, and it was far superior to reference groups in wound re-epithelization and collagen production. CMCS/OHA/Exo hydrogels can promote Exo release based on hydrogel degradation to regulate macrophages transformation and accelerate chronic wound healing. The study offers a method for preparing Exo-loaded hydrogels that effectively promote the transformation of macrophages and accelerate chronic inflammatory wound healing.

Construction and functional evaluation of oral long-acting insulin hydrogel microparticles based on physical and chemical double crosslinking.

The objective of this study was to enhance the convenience and effectiveness of diabetes treatment by developing hydrogel microparticles as an oral insulin delivery system, aiming to reduce the necessity for frequent treatments. The hydrogel microparticles were prepared with polysaccharides through a combination of physical and chemical crosslinking method, they achieved good results in insulin loading efficiency (70 %), insulin release efficiency (98 %) and sustained release time (>20 h). The effective transmembrane transport was validated using an intestinal epithelial cell model, which demonstrated a continuous hypoglycemic effect lasting from 6 to 26 h in a type 2 diabetes mouse model. Additionally, the relative bioavailability of insulin reached 30.14 ± 2.62 %, representing a significant breakthrough in the field of oral insulin delivery carriers. Furthermore, oral insulin hydrogel exhibited a substantial improvement in insulin resistance, organ damage, and diabetes-related complications stemming from hyperglycemia. These compelling findings underscore the potential of hydrogel microparticles as a cost-effective and valuable strategy for oral drug delivery in diabetes treatment.

A Schiff base hydrogel dressing loading extracts from Periplaneta Americana for diabetic wound healing.

As a common complication of diabetic patients, the chronic wound of diabetes has a high incidence, expensive treatment, and recurrence probability, which causes long-term negative impacts on patients' daily life. In this study, the hydrogel was formed by Schiff base reaction between oxidized hyaluronic acid (OHA) and carboxymethyl chitosan (CMCS), and the composite hydrogel dressing was prepared by adding the active polypeptides extract of Periplaneta Americana (PAE). By mass spectrometer determined, PAE mainly includes vitellogenins that can trigger an immune response. The composite hydrogel has good swelling properties, proper fluidity, and a regular 3D network structure. The hydrogel has good cytocompatibility and can promote cell proliferation by L929 fibroblast assay. Finally, it was used to evaluate the effect of diabetic wound repair. The results showed that it could effectively promote wound healing, promote tissue and vascular regeneration, inhibit inflammatory factors, and promote the expression of growth factors. The OHA/CMCS/PAE hydrogels would be promising candidates for chronic wound healing applications.

A composite hydrogel scaffold based on collagen and carboxymethyl chitosan for cartilage regeneration through one-step chemical crosslinking.

The number of cases of cartilage damage worldwide is increasing annually and this problem severely limits an individual's physical activities, subsequently contributing to additional medical problems. Hydrogels can repair cartilage defects and promote cartilage regeneration. In this study, a composite hydrogel scaffold was prepared with collagen (COL), carboxymethyl chitosan (CMC), and the Arg-Gly-Asp (RGD) peptide through one-step chemical crosslinking, in which the three compositions ratio was especially investigated. The hydrogel scaffold performed well in cell adhesion and biocompatibility experiments, mainly due to the favorable porosity (the aperture was concentrated at 100 µm and the porosity was >70 %) and RGD concentration (2 mM RGD was the optimal concentration, which could effectively improve the attachment of BMSCs to the stent). Moreover, bone marrow mesenchymal stem cells (BMSCs) filled in the hydrogel scaffold, together with transforming growth factor TGF-ß3, which was applied to evaluate the feasibility on the repair of the injured cartilage of the rat. In vitro and in vivo study, according to the results of cell proliferation and cytotoxicity, the hydrogel material had no toxic effect on cells, and the COL2/CMC1 hydrogel scaffold had the most obvious role in promoting cell proliferation. The results of pathological section showed that the cell scaffold complex group provided good mechanical properties for the wound and supplemented the stem cells derived from chondrocytes and showed good cartilage defect repair effect; In the scaffold group, the surface fibrosis of the injured area was mainly filled with fibrocartilage and other collagen fibers The hydrogel/BMSCs complex based on COL and CMC can be beneficial for the regeneration of cartilage.

Characterization of polysaccharides from different species of brown seaweed using saccharide mapping and chromatographic analysis.

Brown seaweed polysaccharides (BSPs) are one of the primary active components from brown seaweed that has a range of pharmaceutical and biomedical applications. However, the quality control of BSPs is a challenge due to their complicated structure and macromolecule. In this study, saccharide mapping based on high-performance liquid chromatography (HPLC), multi-angle laser light scattering, viscometer, and refractive index detector (HPSEC-MALLS-Vis-RID), and Fourier transform infrared (FT-IR) were used to discriminate the polysaccharides from nine different species of brown algae (BA1-9). The results showed that BSPs were composed of ß-D-glucans and ß-1,3-1,4-glucan linkages. The molecular weight, radius of gyration, and intrinsic viscosity of BSPs were ranging from 1.718 × 105 Da to 6.630 × 105 Da, 30.2 nm to 51.5 nm, and 360.99 mL/g to 865.52 mL/g, respectively. Moreover, α values of BSPs were in the range of 0.635 to 0.971, which indicated a rigid rod chain conformation. The antioxidant activities of BSPs exhibited substantial radical scavenging activities against DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) radicals, which indicated that the use of BSPs might be a potential approach for antioxidant supplements. Thus, this study gives insights about the structure-function relationship of BSPs, which will be beneficial to improve the quality of polysaccharides derived from marine algae.

Purification and stability analysis of antimicrobial proteins from Varuna litterata.

Ten marine species, including different crabs, bivalve molluscs, and fish intestines were selected to screen the natural antimicrobial protein or peptide as they are enriched with various microorganisms. The crude extract from Varuna litterata, a marine crab which is used as a raw material in the preparation of pickled crabs in Chaoshan area of China, was proved to have a potent bacteriostatic effect against gram-negative bacterium (Escherichia coli) and gram-positive bacterium(Staphylococcus aureus) compared with other marine species. The crude proteins of Varunalitterata were salted-out for preliminary purification and further purified by gel filtration (Sephadex G-150) or anion exchange (DEAE-cellulose 52) chromatographic column. An increase in the antimicrobial activity was noted with the increase in the purity level of the protein. A relatively pure protein was eventually obtained, which was determined to be belonging to the hemocyanin family based on the mass spectrometric data analysis. The purified proteins solution (1 mg/ml) from Varuna litterata exhibited similar antimicrobial activity to that of gentamycin sulfate (0.2 mg/ml), which were relatively stable in a certain pH or temperature range. A structure-activity relationship of the purified hemocyanin was determined based on the interaction of hemocyanin and different chromatographic medium, which revealed that the integrated hexamers played a remarkable role in its bacteriostatic activity. Moreover, the phenoloxidase activity of hemocyanin from Varuna litterata was found as the underlying cause of its antimicrobial potential.

Long-term treatment of polysaccharides-based hydrogel microparticles as oral insulin delivery in streptozotocin-induced type 2 diabetic mice.

To develop a more effective and safer drug for the treatment of type 2 diabetes mellitus (T2DM), polysaccharides-based hydrogel microparticles as oral insulin delivery was prepared and explored. This study was aimed to evaluate the antidiabetic effects and hypoglycemic mechanism with long-term administration(four weeks) of oral insulin hydrogel microparticles in type 2 diabetic mice on a model of diabetes using a high fat diet combined with streptozotocin. The results revealed that the long-term treatment of oral insulin polysaccharides-based hydrogel microparticles could significantly alleviate the symptoms of polyphagia, polydipsia, polyuria and weight loss in diabetic mice. Also, oral administration of insulin hydrogel microparticles could significantly reduce fasting blood glucose levels, ameliorate insulin resistance and increase insulin sensitivity in the mice with T2DM. The concentration of plasma TG, TC, LDL-C, FFA, BUN, CRE significantly decreased and the levels of HDL-C increased showed that insulin polysaccharides-based hydrogel microparticles were effective in regulating lipid metabolism and prevent diabetic nephropathy complication in diabetic mice. In addition, the supplementation of insulin hydrogel microparticles could significant improve the antioxidant capacity by increasing the level of SOD, CAT and decreasing the level of MDA, GPT, NO, TNF-α, and reverse histological deterioration of kidney and pancreas in diabetic mice. The above outcome concluded that insulin polysaccharides-based hydrogel microparticles may exhibit promising anti-diabetic activity and the potential to be a drug candidate for T2DM.

Carboxymethyl ß-cyclodextrin grafted carboxymethyl chitosan hydrogel-based microparticles for oral insulin delivery.

This study was aimed at utilizing polysaccharides for the development of effective hydrogel microparticles for oral insulin delivery that has a controlled, and sustained release to enhance paracellular transcellular absorption. Carboxymethyl ß-cyclodextrin grafted carboxymethyl chitosan hydrogels (CMCD-g-CMCs) were prepared from carboxymethyl ß-cyclodextrin (CMCD) and carboxymethyl chitosan (CMC) using a water-soluble carbodiimide as a crosslinker in the presence of N-hydroxysuccinimide. After synthesis, the hydrogel structures were determined via FT-IR and XRD analyses. The porous structure of hydrogels was confirmed by SEM observations and swelling behaviours. The insulin release behaviours were found to betriggered by pH in vitro. Results showed that insulin was successfully retained inside the hydrogels in the gastric environment and slowly released following passage to intestinal conditions. The stability of the secondary structure of insulin was studied by dichroism circular (CD) and fluorescence (FL) spectrophotometer measurement. There was no significant difference in the secondary structure between the native and released insulin. In vitro studies revealed that the hydrogel microparticles exhibited non-cytotoxicity and were transported across the Caco-2 cell monolayers mainly via the paracellular pathway. In order to examine the effectiveness of hydrogel-based sustained release microparticles in delivering insulin in vivo, we administered different insulin-loaded hydrogel microparticles to diabetic mice. In these studies, we found that the insulin-loaded hydrogel microparticles provided a significant and sustained (ranging from 6 h to 12 h) reduction in the blood glucose levels of diabetic mice compared with subcutaneous injection. Overall, these findings demonstrate that CMCD-g-CMCs may be a promising protein carrier for use in oral drug delivery.