chondroitin and Cicatrix

The precise contribution of different CS-GAGs to CSPG-mediated inhibition of axonal growth after CNS injury is unknown. Quantification of the CS-GAGs in uninjured and injured brain (scar tissue) using fluorophore-assisted carbohydrate electrophoresis (FACE) demonstrated that the dominant CS-GAG in the uninjured brain is CS-4 whereas, in glial scar, CS-2, CS-6, and CS-4,6 were over-expressed. To determine if the pattern of sulfation influenced neurite extension, we compared the effects of CS-GAGs with dominant CS-4, CS-6, or CS-4,6 sulfation to intact CSPG (aggrecan), chondroitin (CS-0), and hyaluronan on chick DRG neurite outgrowth. We report that CS-4,6 GAG, one of the upregulated CS-GAGs in astroglial scar, is potently inhibitory and is comparable to intact aggrecan, a CSPG with known inhibitory properties. Thus, a specific CS-GAG that is differentially over-expressed in astroglial scar is a potent inhibitor of neurite extension. These results may influence the design of more specific strategies to enhance CNS regeneration after injury.

Topics: Aggrecans; Animals; Brain Injuries; Chick Embryo; Chondroitin; Chondroitin Sulfates; Cicatrix; Extracellular Matrix Proteins; Female; Ganglia, Spinal; Gliosis; Growth Cones; Growth Inhibitors; Hyaluronic Acid; Lectins, C-Type; Neurites; Proteoglycans; Rats; Sulfates; Up-Regulation

A D-glucuronic acid rich, copolymeric chondroitin sulfate (CS)-dermatan sulfate (DS) proteoglycan (PG) from post-burn hypertrophic scar tissue (HSc) was obtained by DEAE-cellulose chromatography and differential ethanol fractionation, and further purified on a Sepharose CL-6B column. CS-DS-PG protein content was 14% (w/w). The amino-terminal amino acid sequence of the first ten residues was as follows: NH2-Asp-Glu-Ala-B-Gly-Ile-Gly-Pro-Glu-Val. This sequence is identical to that of human embryonic fibroblast cell (IMR-90) CS-DS-PG, as well as to human HSc-DS-PG. After chondroitinase ABC treatment, two peptides (Mr 22,000 and 16,000 daltons) were detected by sodium dodecyl sulfate-(polyacryl)amide gel electrophoresis (SDS-PAGE). ELISA analysis using rabbit antiserum raised against a synthetic peptide that contained 15 amino acids in the same sequence as the amino terminus of human fetal membrane PG showed significant reactivity with HSc CS-DS-PG. HSc CS-DS-PG had an apparent Mr of approximately 78,000 daltons, as determined by Sepharose CL-6B chromatography and SDS-PAGE. Alkaline borohydride treatment of CS-DS-PG liberated CS-DS glycosaminoglycan (GAG) chains having an Mr of 29,000 daltons. The conversion of xylose to xylitol indicated that the GAG chains are attached to the PG protein core at O-3 through a xylosyl-seryl linkage. CS-DS-PG also contained both N and O-linked oligosaccharides and did not aggregate with hyaluronic acid. These results, together with those reported previously, showed that HSc CS-DS-PG and DS-PG have the same A1-A15 amino acid sequence at the amino terminus but different protein cores. HSc CS-DS-PG was completely digested with chondroitinase AC and is, therefore, distinctly different from HSc DS-PG.

Topics: Aggrecans; Amino Acid Sequence; Amino Acids; Animals; Burns; Carbohydrates; Chondroitin; Chondroitin Sulfate Proteoglycans; Cicatrix; Dermatan Sulfate; Extracellular Matrix Proteins; Glycoproteins; Humans; Lectins, C-Type; Molecular Sequence Data; Proteoglycans; Skin

An experimental investigation of the ability of the chondroitinsulphate preparation produced from cattle tracheas, of rumalon and chonsuride to stimulate the regeneration of cutaneous coverings in case of their injury was carried out. A strength of the surgical cicatrix and its elongation at rupture in two weeks' injections of chondroitinsulphate increased in comparison with control. A stimulating effect of chondroitinsulphate to regeneration of flesh wound in case of local single action didn't differ essentially from the effect of chonsuride. In case of application of the preparations, an area of wound for the rats received chondroitinsulphate was already on the eighth day twice as less in comparison with the animals treated with chonsuride. Thus, a stimulating effect of chondroitinsulphate preparation to the regeneration of damaged cutaneous coverings by both parenteral and local administration was shown.

Topics: Animals; Chondroitin; Chondroitin Sulfates; Cicatrix; Drug Evaluation, Preclinical; Male; Rats; Rats, Inbred Strains; Stimulation, Chemical; Time Factors; Tissue Extracts; Wound Healing

Dermatan sulfate (DS) proteoglycans (PGs) were extracted from human post-burn scar (Sc) tissues with 4M guanidinium chloride and isolated from the extracts by DEAE-cellulose chromatography and by differential ethanol precipitation. The DS.PGs were further purified by Sepharose CL-6B column chromatography. The average molecular weight (Mr) of hypertrophic scar (HSc) tissue DS.PGs was 39,000 based on sedimentation equilibrium measurements. Alkaline borohydride treatment of DS.PGs liberated glycosaminoglycan (GAG) chains and the presence of xylitol indicated that these chains were attached to protein core by xylosyl residues. The average Mr of the DS.GAG chain from HSc and normal scar (NSc) samples were 23,500 and 20,000 respectively. After digestion of the HSc and NSc, DS.PGs with chondroitinase ABC in the presence of proteinase inhibitors, two peptide components with Mr values of 21,500 and 17,000 were detected by SDS-polyacrylamide gel electrophoresis using reducing conditions. Analysis of the protein core fractions derived from NSc and HSc DS.PGs by Sepharose CL-6B column chromatography showed the presence of a single NH2-terminal amino acid (aspartic acid) and also that the fractions with different KAV values had an identical NH2-terminal sequence (A1-A5). The A1-A23 sequence of NSc DS.PG (major fraction, C): NH2Asp-Glu-Ala-O-Gly-Ile-Gly-Pro-Glu-Val-Pro-Asp-Asp-Arg-Asp-Phe-G lu-Pro- Ser-Leu-Gly-Pro-Val was the same as reported for a DS.PG isolated from human fetal membrane (HFM) tissue (Brennan et al., 1984). ELISA inhibition assay using monoclonal antibodies raised in rabbit against the NH2-terminal peptide (containing 15 amino acids) of human fetal membrane tissue were found to cross-react with HSc and NSc DS.PGs. Monoclonal antibodies to bovine skin DS.PGs protein core (Pearson et al., 1983) did not show any cross-reactivity with scar DS.PGs. These results show that the scar DS.PGs described here are different from normal bovine skin DS.PGs in the size and type of the protein core, and that in all the samples, the peptide components have the same NH2-terminal amino acid sequence.

Topics: Amino Acids; Animals; Burns; Carbohydrates; Chondroitin; Chondroitin Sulfate Proteoglycans; Chromatography, DEAE-Cellulose; Chromatography, Gel; Cicatrix; Dermatan Sulfate; Glycosaminoglycans; Humans; Molecular Weight; Proteoglycans

Topics: Adult; Breast; Chondroitin; Chondroitin Sulfates; Cicatrix; Connective Tissue; Cytoskeleton; Female; Fibroblasts; Glycosaminoglycans; Histocytochemistry; Humans; Microscopy, Electron; Middle Aged; Prostheses and Implants

Topics: Biopsy; Chondroitin; Cicatrix; Collagen; Dermatan Sulfate; Glycosaminoglycans; Granulation Tissue; Humans; Hyaluronic Acid; Microscopy, Electron; Microscopy, Electron, Scanning; Ruthenium; Skin; Water

Topics: Acetates; Adolescent; Adult; Aged; Burns; Cellulose; Child; Child, Preschool; Chondroitin; Cicatrix; Electrophoresis; Galactosamine; Glucosamine; Glycosaminoglycans; Hexosamines; Humans; Hyaluronic Acid; Hyaluronoglucosaminidase; Lyases; Middle Aged; Skin

Topics: Amino Sugars; Animals; Chondroitin; Cicatrix; Connective Tissue; Edema; Glycoproteins; Glycosaminoglycans; Heart Injuries; Hexosamines; Hyaluronic Acid; Isoproterenol; Male; Myocardium; Necrosis; Rats; Wound Healing

Topics: Chondroitin; Cicatrix; Glycosaminoglycans; Humans; Hyaluronic Acid; Hydrogen-Ion Concentration; Solubility; Sulfates; Tampons, Surgical