title: Glycosaminoglycan metabolism in otosclerotic bone cells.
abstract: Normal and otosclerotic bone cells were cultured in vitro in serum-free medium to evaluate single glycosaminoglycan (GAG) class synthesis and secretion. Moreover, the degradative process was studied by inhibiting the lysosomal functions through the addition of ammonium chloride to the cultures, an ammine known to inhibit lysosomal degradation by neutralizing organelle activity. Otosclerotic bone cells accumulated a lower amount of GAG both in the cellular and extracellular pool compared to normal ones. The decrease was markedly higher for secreted GAG. Moreover a different pattern of single GAG class distribution was observed in the two cell types considered. In the medium of otosclerotic cells a percentage increase of hyaluronic acid (HA) and dermatan sulphate (DS) and a percentage decrease of heparan sulfate (HS) and chondroitin sulfate (CS) were observed compared to normal bone cells. Ammonium chloride had a lower effect on pathologic than on normal cells, indicating a decrease in the degradative process in otosclerotic bone cells. These results were also confirmed by the experiments on GAG uptake and degradation and by the dosage of enzymatic activity of two exoglycosidases. Since extracellular GAG composition influences bone deposition and mineralization, these data support the hypothesis that otosclerosis is the result of an error in the connective tissue matrix structure.
authors.affiliation: Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università di Perugia, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Locci
authors.forename: E
authors.initials: E
authors.lastname: Becchetti
authors.forename: G
authors.initials: G
authors.lastname: Venti
authors.forename: C
authors.initials: C
authors.lastname: Lilli
authors.forename: L
authors.initials: L
authors.lastname: Marinucci
authors.forename: E
authors.initials: E
authors.lastname: Donti
authors.forename: G
authors.initials: G
authors.lastname: Paludetti
authors.forename: M
authors.initials: M
authors.lastname: Maurizi
mesh_terms: D000118:Acetylglucosaminidase / Q000378:metabolism; D000643:Ammonium Chloride / Q000494:pharmacology; D001692:Biological Transport / Q000502:physiology; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D005260:Female; D005944:Glucosamine / Q000378:metabolism; D005966:Glucuronidase / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology; D010040:Otosclerosis / Q000201:enzymology / Q000473:pathology*; D014316:Tritium
chemical_list: D006025:Glycosaminoglycans; D000643:Ammonium Chloride; D014316:Tritium; D006026:Glycoside Hydrolases; D005966:Glucuronidase; D000118:Acetylglucosaminidase; D005944:Glucosamine