chondroitin-sulfates and cellulose-sulfate

Cartilage tissue engineering strategies seek to repair damaged tissue using approaches that include scaffolds containing components of the native extracellular matrix (ECM). Articular cartilage consists of glycosaminoglycans (GAGs) which are known to sequester growth factors. In order to more closely mimic the native ECM, this study evaluated the chondrogenic differentiation of mesenchymal stem cells (MSCs), a promising cell source for cartilage regeneration, on fibrous scaffolds that contained the GAG-mimetic cellulose sulfate. The degree of sulfation was evaluated, examining partially sulfated cellulose (pSC) and fully sulfated cellulose (NaCS). Comparisons were made with scaffolds containing native GAGs (chondroitin sulfate A, chondroitin sulfate C and heparin). Transforming growth factor-beta3 (TGF-β3) sequestration, as measured by rate of association, was higher for sulfated cellulose-containing scaffolds as compared to native GAGs. In addition, TGF-β3 sequestration and retention over time was highest for NaCS-containing scaffolds. Sulfated cellulose-containing scaffolds loaded with TGF-β3 showed enhanced chondrogenesis as indicated by a higher Collagen Type II:I ratio over native GAGs. NaCS-containing scaffolds loaded with TGF-β3 had the highest expression of chondrogenic markers and a reduction of hypertrophic markers in dynamic loading conditions, which more closely mimic in vivo conditions. Studies also demonstrated that TGF-β3 mediated its effect through the Smad2/3 signaling pathway where the specificity of TGF-β receptor (TGF- βRI)-phosphorylated SMAD2/3 was verified with a receptor inhibitor. Therefore, studies demonstrate that scaffolds containing cellulose sulfate enhance TGF-β3-induced MSC chondrogenic differentiation and show promise for promoting cartilage tissue regeneration.

Topics: Cartilage, Articular; Cellulose; Chondrogenesis; Chondroitin Sulfates; Glycosaminoglycans; Tissue Scaffolds; Transforming Growth Factor beta; Transforming Growth Factor beta3

Articular cartilage has limited healing and self-repair capability. Damage to articular cartilage becomes irreversible leading to osteoarthritis, which can impact a person's quality of life. Approximately, 5-10% of cartilage tissue is made up of sulfated glycosaminoglycans (GAGs), which sequester growth factors as well as provide structural integrity to the native cartilage tissue. This study evaluated the chondrogenic differentiation of human mesenchymal stem cells (MSCs) on gelatin-based scaffolds containing partially sulfated cellulose (pSC), a GAG mimetic derived from cellulose, in comparison to native GAGs, chondroitin sulfate-A (CS-A) and chondroitin sulfate-C (CS-C), where pSC has similarity to CS-C in terms of degree and pattern of sulfation. Scaffolds were prepared by electrospinning gelatin with pSC or the native GAGs. All scaffolds consist of fibers having average diameters of approximately 3 μm and inter-fiber spacing of approximately 30 μm and were hydrolytically stable throughout the culture. MSCs cultured on pSC containing scaffolds showed early production of sulfated GAGs and higher collagen type II to type I ratio than native GAGs. Among the native GAGs, chondrogenesis was promoted to a greater extent for CS-C in comparison to CS-A containing scaffolds, which suggests the pattern of sulfation impacts chondrogenesis. Partially sulfated cellulose could be used as a potential GAG mimic for cartilage tissue engineering applications.

Topics: Biomimetic Materials; Cartilage, Articular; Cell Differentiation; Cells, Cultured; Cellulose; Chondrogenesis; Chondroitin Sulfates; Collagen; Extracellular Matrix; Glycosaminoglycans; Humans; Mesenchymal Stem Cells; Tissue Engineering; Tissue Scaffolds

Spinal cord injury can lead to severe dysfunction as a result of limited nerve regeneration that is due to an inhibitory environment created at the site of injury. Neural tissue engineering using materials that closely mimic the extracellular matrix (ECM) during neural development could enhance neural regeneration. Glycosaminoglycans (GAGs), which are sulfated polysaccharides, have been shown to modulate axonal outgrowth in neural tissue depending upon the position and degree of sulfation. Cellulose sulfate (CelS), which is a GAG mimetic, was evaluated for its use in promoting neurite extension. Aligned fibrous scaffolds containing gelatin blended with 0.25% partially sulfated cellulose sulfate (pCelS), having sulfate predominantly at the 6-carbon position of the glucose monomer unit, and fully sulfated cellulose sulfate (fCelS), which is sulfated at the 2-, 3-, and 6-carbon positions of the glucose monomer unit, were fabricated using the electrospinning method. Comparisons were made with scaffolds containing native GAGs, chondroitin sulfate-A (CS-A) and chondroitin sulfate-C (CS-C), which were obtained from commercial sources. CS-A and CS-C are present in neural tissue ECM. The degree of sulfation and position of sulfate groups was determined using elemental analysis, Fourier-transform infrared spectroscopy (FTIR), Raman microspectroscopy, and

Topics: Animals; Biomimetic Materials; Cellulose; Chondroitin Sulfates; Neurites; Neurogenesis; Rats; Rats, Sprague-Dawley; Tissue Scaffolds

Polyanions (in an amount within 1.5 - 6.0 mg), including cellulose sulfates (excreted from Gossipium hirsutum L., molecular weight 22.0 kDa, degree of sulfation within 0.8 - 1.8), inulin sulfates (excreted from Helianthus tuberosus, molecular weight 8.0 kDa, degree of sulfation within 0.6 - 1.6), pectin sulfates (excreted from Abies sibirica L., molecular weight 24.0 kDa, degree of sulfation within 0.8 - 1.1), give rise to peaks of precipitation with polycations of protamine sulfate. Only cellulose sulfates (in amount within 0.38 - 6.00 mg) give the peaks of precipitation with chitosan polycations (molecular weight 10 kDa, degree of deacetylation 85%) during horizontal biospecific electrophoresis. The height of the peak of precipitation with protamin sulfate was found to grow with increasing antithrombin activity of cellulose sulfates and pectin sulfate (for polyanions in an amount within 1.5 - 6 mg). The size of the area of precipitation with chitosan was found to decrease with increasing antithrombin activity of cellulose sulfates.

Topics: Anticoagulants; Blood Coagulation; Cellulose; Chondroitin Sulfates; Electrophoresis, Agar Gel; Gossypium; Humans; Inulin; Molecular Weight; Pectins; Pinus; Polyamines; Polyelectrolytes; Protamines; Thrombin

The reduction of acute toxicity of paraquat dichloride (PQ) in mice was studied by several sodium sugar sulfates such as dextran sulfate (DS), cellulose sulfate (CS), chondroitin sulfate (CDS), sucrose sulfate (SS) and glucose sulfate (GS). When sugar sulfates (DS, CS, SS or GS, 2,000 mg/kg) were given orally immediately after PQ ingestion (200 mg/kg), the survival rates were 100% respectively on the 14th day after PQ ingestion. In the case of CDS, it was 73%. The effective dose of SS and GS given for the prevention of PQ toxicity in mice after PQ (200 mg/kg) ingestion was more than 400 mg/kg for SS-treated group and more than 800 mg/kg for GS-treated group. The survival rate of these mice was 100% respectively. When GS or SS (2,000 mg/kg) was given orally to mice within 1 h (0, 10, 20, 30 or 60 min) after PQ ingestion (200 mg/kg), the survival rates in an earlier treatment were greater than those in a later treatment. The measurement of serum PQ concentrations in mice suggested that a mechanism for the prevention of PQ toxicity by the administration of sugar sulfates may be inhibition of PQ absorption, and/or stimulation of PQ excretion from intestine. The effectiveness of SS and GS in preventing PQ toxicity was similar to that of DS. These results suggested that SS, GS and DS might serve as an antidote for acute toxicity of PQ.

Topics: Animals; Carbohydrates; Cellulose; Chondroitin Sulfates; Dextran Sulfate; Dextrans; Glucose; Lethal Dose 50; Male; Mice; Mice, Inbred Strains; Molecular Weight; Organometallic Compounds; Paraquat; Sucrose; Sulfates

In our previous paper, we reported that various types of carrageenan, dextran sulfate and fucoidan, which are sulfated homopolysaccharides with high molecular weights, were human T cell mitogens and murine polyclonal B cell activators (PBAs) and that heparin, a sulfated heteropolysaccharide, was a very weak human mitogen and mouse PBA. Here we used cellulose sulfate (Mr 7-9 X 10(3], dextran sulfate with two different low molecular weights (Mr 5 X 10(3) and 8 X 10(3], two different condroitin sulfates (Mr 3.5 X 10(4], polyvinyl sulfate and polygalacturonic acid to investigate mitogenic activities of polysaccharides in detail. The following results were obtained. Low-molecular-weight sulfated homopolysaccharides, dextran sulfate and cellulose sulfate, were very weak or not human T cell mitogens. However, they were better murine PBAs. Sulfated heteropolysaccharides, chondroitin 4-sulfate and chondroitin 6-sulfate, hardly induced mitogenic changes in human T cells and mouse B cells, even though the molecular weight of these substances was more than 1 X 10(4). There were no other polymers examined so far which activated both human T cells and murine B cells. The relationship among molecular size, sulfate groups and lymphocyte activation is discussed in detail.

Topics: Adult; Animals; B-Lymphocytes; Cellulose; Chondroitin Sulfates; Dextran Sulfate; Dextrans; Female; Humans; In Vitro Techniques; Lymphocyte Activation; Male; Mice; Mitogens; Molecular Weight; Pectins; Polysaccharides; Polyvinyls; T-Lymphocytes