chondroitin-sulfates and Diabetes-Mellitus

Improving chronic wound healing remains a challenge in the clinical practice. In this study, we developed double-crosslinked angiogenic 3D-bioprinted patches for diabetic wound healing by the photocovalent crosslinking of vascular endothelial growth factor (VEGF) using ultraviolet (UV) irradiation. 3D printing technology can precisely customize the structure and composition of patches to meet different clinical requirements. The biological polysaccharide alginate and chondroitin sulfate methacryloyl were used as biomaterials to construct the biological patch, which could be crosslinked using calcium ion crosslinking and photocrosslinking, thereby improving its mechanical properties. More importantly, acrylylated VEGF could be easily and rapidly photocrosslinked under UV irradiation, which simplified the step of chemically coupling growth factors and prolonged VEGF release time. These characteristics suggest that 3D-bioprinted double-crosslinked angiogenic patches are ideal candidates for diabetic wound healing and other tissue engineering applications.

Topics: Alginates; Chondroitin Sulfates; Diabetes Mellitus; Hydrogels; Printing, Three-Dimensional; Tissue Engineering; Tissue Scaffolds; Vascular Endothelial Growth Factor A; Wound Healing

Topics: Alginates; Chondroitin Sulfates; Diabetes Mellitus; Humans; Hydrogels; Spectroscopy, Fourier Transform Infrared; Wound Healing

Hypercholesterolemia is one of the factors contributing to cardiovascular problems. Erythrocytes are known to contribute its cholesterol to atherosclerotic plaque. Our earlier study showed that erythrocytes overexpress chondroitin sulphate/dermatan sulphate (CS/DS), a linear co-polymer, during diabetes which resulted in increased cytoadherence to extracellular matrix (ECM) components. This study was carried out to determine whether diet-induced hypercholesterolemia had any effect on erythrocyte CS/DS and impacted cytoadherence to ECM components. Unlike in diabetes, diet-induced hypercholesterolemia did not show quantitative changes in erythrocyte CS/DS but showed difference in proportion of un-sulphated and 4-O-sulphated disaccharides. Erythrocytes from hypercholesterolemic rats showed increased adhesion to ECM components which was abrogated to various extents when subjected to chondroitinase ABC digestion. However, isolated CS/DS chains showed a different pattern of binding to ECM components indicating that orientation of CS/DS chains could be playing a role in binding.

Topics: Animals; Cell Adhesion; Chondroitin Sulfates; Dermatan Sulfate; Diabetes Mellitus; Erythrocytes; Extracellular Matrix; Humans; Hypercholesterolemia; Rats; Structure-Activity Relationship

In diabetes the endothelium is either chronically or transiently exposed to hyperglycemic conditions. In addition, endothelial dysfunction in diabetes is related to changes in the inflammatory response and the turnover of extracellular matrix. This study was undertaken to study the effects of inflammatory stimuli on one particular matrix component, the heparan sulfate (HS) proteoglycans (PGs) synthesized by primary human umbilical cord vein endothelial cells (HUVEC). Such cells were cultured in vitro in 5 mM and 25 mM glucose. The latter concentration was used to mimic hyperglycemic conditions in short-term experiments. HUVEC were also cultured in the presence of the inflammatory agents tumor necrosis factor α (TNF-α), interleukin 1α (IL-1α), interleukin 1β (IL-1β) and transforming growth factor β (TGF-β). The cells were labeled with (35)S-sulfate and (35)S-PGs were recovered for further analyses. The major part of the (35)S-PGs was secreted to the medium, irrespective of type of stimuli. Secreted (35)S-PGs were therefore isolated and subjected to further analyses. TNF-α and IL-1α slightly increased the release of (35)S-PGs to the culture medium, whereas IL-1β treatment gave a significant increase. The different treatments neither changed the ratio of (35)S-HS and (35)S-chondroitin sulfate (CS) nor the macromolecular properties of the (35)S-PGs. However, the (35)S-HS chains were slightly increased in size after TNF-α treatment, and slightly decreased after TGF-β treatment, but not affected by the other treatments. Compositional analysis of labeled disaccharides showed changes in the amount of 6-O-sulfated glucosamine residues after treatment with TNF-α, IL-1α and IL-1β. Western immunoblotting showed that major HSPGs recovered from these cells were collagen XVIII, perlecan and agrin, and that secretion of these distinct PGs was increased after IL-1β stimulation. Hence, short term inflammatory stimuli increased the release of HSPGs in HUVEC and affected both the size and sulfation pattern of HS, depending on type of stimuli.

Topics: Agrin; Cells, Cultured; Chondroitin Sulfates; Collagen Type XVIII; Cytokines; Diabetes Mellitus; Endothelium; Extracellular Matrix; Glucosamine; Glucose; Heparan Sulfate Proteoglycans; Human Umbilical Vein Endothelial Cells; Humans; Hyperglycemia; Interleukin-1alpha; Interleukin-1beta; Sulfur Radioisotopes; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

The biomechanical changes in rabbit cornea preserved in storage media with different glucose concentrations are experimentally assessed. Two groups of eight fresh rabbit corneas were preserved for 10 days in storage medium Optisol-GS with glucose concentrations of 14 and 28 mM, respectively. Eight additional corneas preserved, glucose-free, in the same medium served as the control group. All specimens were tested under inflation conditions up to 45 mmHg posterior pressure, and the pressure-deformation data obtained experimentally were analyzed using shell theory to derive the stress-strain behavior. Comparisons were held between the three specimen groups in order to determine the effect of glucose concentration on corneal biomechanical behavior and thickness. After storage, the mean central corneal thickness in the control, low-glucose and high-glucose groups underwent statistically significant increases of 38.7 ± 11.3%, 45.4 ± 7.6% and 50.6 ± 8.6%, respectively. The corneas also demonstrated consistent stiffness increases with higher glucose concentrations. The tangent modulus values determined at different pressure levels between 10 and 40 mmHg underwent statistically significant increases with glucose level (P < 0.05). Compared to the control group, other specimens had higher tangent modulus by 17-20% on average with low glucose and 30-37% with high-glucose concentration. The results of the study indicate that the influence of the high-glucose level commonly experienced in diabetes on the biomechanical stiffness of the cornea should be considered in clinical management and in understanding corneal ectasia, glaucoma and the response to refractive surgery.

Topics: Animals; Biomechanical Phenomena; Chondroitin Sulfates; Complex Mixtures; Cornea; Cryopreservation; Dextrans; Diabetes Mellitus; Elasticity; Gentamicins; Glucose; Intraocular Pressure; Models, Theoretical; Organ Preservation; Organ Preservation Solutions; Rabbits; Transducers, Pressure

Characterization of the analytic profile of proteoglycans in the intervertebral discs at L4-L5 of nondiabetic (n = 5) and diabetic (n = 5) age-matched subjects. The discs used were discarded material from operations.. To clarify the reason for the higher risk of disc prolapse in diabetic patients.. The pathogenesis of diabetes results from a combination of neurologic dysfunctions and a yet undefined metabolic failure, which leads to an abnormal proteoglycan profile.. The following methods were used to determine the proteoglycan profile: the measurement of 35S-sulfate uptake per gram wet tissue into sulfated glycosaminoglycan using fresh tissue explants; extraction of proteoglycans by 4 M guanidinium chloride containing protease inhibitors, with further purification by ultracentrifugation on cesium chloride buoyant density gradient under dissociative conditions; total uronic acid and protein contents in the various gradient fractions; assessing the length of sugar side chains of isolated 35Sulfate-glycosaminoglycan molecules by separation of the glycosaminoglycan molecules on a Sepharose 6B-CL column; and paper chromatography of the final digest products of glycosaminoglycan molecules obtained by chondroitinase ABC, a glycosaminoglycan-degrading enzyme.. The findings show that discs from normal nondiabetic subjects have 15 times the rate of 35Sulfate incorporation into glycosaminoglycan molecules than do discs of diabetic patients. The proteoglycans of diabetic patients are banded at a lower buoyant density, indicating a lowered glycosylation rate and a lower number of sugar side chains per core protein. In discs of diabetic patients, there is a slight increase in the chain length of chondroitin sulfate. Further analysis of the glycosaminoglycan chains showed a decreased amount of keratan sulfate, compared with that in nondiabetic subjects. However, the total uronic acid content of the disc tissues and the ratio of uronic acid to protein of each fraction were unchanged in diabetic patients versus that in control subjects.. Discs in patients with diabetes have proteoglycans with lower buoyant density and substantially undersulfated glycosaminoglycan, which with the specific neurologic damage in these patients, might lead to increased susceptibility to disc prolapse.

Topics: Aged; Chondroitin Sulfates; Chromatography, Gas; Diabetes Complications; Diabetes Mellitus; Female; Glycosaminoglycans; Humans; Intervertebral Disc; Intervertebral Disc Displacement; Keratan Sulfate; Low Back Pain; Lumbar Vertebrae; Male; Middle Aged; Proteoglycans; Uronic Acids

Topics: Child; Chondroitin Sulfates; Diabetes Mellitus; Glomerulonephritis; Glycosaminoglycans; Heparitin Sulfate; Humans; Mucopolysaccharidoses; Nephritis, Hereditary; Nephrotic Syndrome; Proteinuria

Addition of heparin and dextran sulphate to human skin fibroblasts in cell cultures caused an increase in [3H]-proline incorporation into collagen and total protein in the culture medium by cells derived from nondiabetics. Cells from type 2 diabetic subjects were significantly less affected by dextran sulphate addition, suggesting altered regulatory mechanisms for collagen production in these cells. Addition of chondroitin sulphate caused a dose-dependent increase in labelled collagen, indicating a possible role for this glycosaminoglycan as modulator of collagen deposition.

Topics: Adult; Aged; Cells, Cultured; Chondroitin Sulfates; Collagen; Dextran Sulfate; Dextrans; Diabetes Mellitus; Female; Fibroblasts; Heparin; Humans; Insulin; Male; Middle Aged; Protein Biosynthesis; Skin

The serum concentrations of different glycosaminoglycan fractions have been studied in diabetic subjects without evidence of vascular complications and compared with those of age-and sex-matched normal subjects. Electrophoretic analysis of the various glycosaminoglycan fractions showed an increase in hyalunoric acid and heparin, and a decrease in heparan sulfate, chondroitin-4-sulfate and chondroitin-6-sulfate. Furthermore, the levels of glycosylated haemoglobin A1 were positively correlated with those of hyaluronic acid (r = 0.96, p. less than 0.001) and heparin (r = 0.45, p less than 0.01) in the diabetic group. The present data suggest thah a) in diabetics there is a shift in the metabolism of glycosaminoglycans towards a decreased sulfated glycosaminoglycans/hyaluronic acid ratio; and b) long-lasting hyperglycemia is correlated with the disturbances in the metabolism of glycosaminoglycans.

Topics: Adult; Chondroitin Sulfates; Diabetes Mellitus; Female; Glycated Hemoglobin; Glycosaminoglycans; Heparin; Heparitin Sulfate; Humans; Hyaluronic Acid; Male

Cultured fibroblasts derived from normal subjects and juvenile diabetics attach in the absence of serum to plastic culture dishes and secrete macromolecules, including collagenous components, hyaluronic acid, and proteoglycans into the medium and onto the plastic surface where they form a microexudate carpet. Most diabetic fibroblasts examined did not spread as well as normal cells during a 4-hr interval after the initial attachment. There were no significant differences between normal and diabetic cells with respect to proline and lysine incorporation and lysine hydroxylation. The percentage glycosylation of hydroxylysine was marginally higher in the media proteins of diabetic cells, but glycosylation in both normal and diabetic cells was elevated over that typically observed in human skin collagen. Collagenous components were estimated to constitute approximately 15-20% of the microexudate carpet fraction in both normal and diabetic cell strains. Diabetic fibroblasts exhibited a marginally lower ratio of heparan sulfate to chondroitin sulfate in the cell surface to matrix microexudate carpet fraction (trypsinate) than did normal fibroblasts. The hyaluronate and chondroitin sulfate contents of this fraction of diabetic cells were not significantly different from those of normal cells.

Topics: Cell Adhesion; Cell Line; Chondroitin Sulfates; Collagen; Diabetes Mellitus; Fibroblasts; Glycosaminoglycans; Heparitin Sulfate; Humans; Lysine; Proline

Topics: Chondroitin Sulfates; Diabetes Mellitus; Drug Combinations; Heparin; Humans

Topics: Chondroitin; Chondroitin Sulfates; Diabetes Complications; Diabetes Mellitus; Disease; Humans; Retina; Retinal Diseases