title: Effect of glucosamine and its peptidyl-derivative on the production of extracellular matrix components by human primary chondrocytes.
abstract: OBJECTIVE
  Aim of this study is to investigate the effects of Glucosamine (GlcN) and its peptidyl-derivative, 2-(N-Acetyl)-L-phenylalanylamido-2-deoxy-β-D-glucose (NAPA), on extracellular matrix (ECM) synthesis in human primary chondrocytes (HPCs).
  
  
  METHODS
  Dose-dependent effect of GlcN and NAPA on Glycosaminoglycan (GAG), Collagen type II (Col2) and Small Leucine-Rich Proteoglycans (SLRPs) was examined by incubating HPCs, cultured in micromasses (3D), with various amounts of two molecules, administered as either GlcN alone or NAPA alone or GlcN plus NAPA (G + N). Immunohystochemical and immunofluorescent staining and biochemical analysis were used to determine the impact of the two molecules on ECM production. Gene expression analysis was performed by TaqMan Real-Time Polymerase Chain Reaction (PCR) assays.
  
  
  RESULTS
  The lowest concentration to which GlcN and NAPA were able to affect ECM synthesis was 1 mM. Both molecules administered alone and as G + N stimulated GAGs and SLRPs synthesis at different extent, NAPA and mainly G + N stimulated Col2 production, whereas GlcN was not effective. Both molecules were able to induce Insulin Growth Factor-I (IGF-I) and to stimulate SOX-9, whereas NAPA and G + N were able to up-regulate both Hyaluronic Acid Synthase-2 and Hyaluronic acid. Very interesting is the synergistic effect observed when chondrocyte micromasses were treated with G + N.
  
  
  CONCLUSIONS
  The observed anabolic effects and optimal concentrations of GlcN and NAPA, in addition to beneficial effects on other cellular pathways, previously reported, such as the inhibition of IKKα, could be useful to formulate new cartilage repair strategies.
authors.affiliation: Dept. of Biochemical Sciences, Sapienza University of Roma, P.le Aldo Moro, 5, 00185 Roma, Italy. Electronic address: daniela.stoppoloni@uniroma1.it.
authors.forename: D
authors.initials: D
authors.lastname: Stoppoloni
authors.affiliation: Dept. of Biochemical Sciences, Sapienza University of Roma, P.le Aldo Moro, 5, 00185 Roma, Italy. Electronic address: laura.politi@uniroma1.it.
authors.forename: L
authors.initials: L
authors.lastname: Politi
authors.affiliation: Dept. of Medico-Surgical Sciences and Biotechnologies, Faculty of Medicine and Pharmacy, Sapienza University, Polo Pontino, Corso Della Repubblica 79, Latina, Italy. Electronic address: m.leopizzi@hotmail.it.
authors.forename: M
authors.initials: M
authors.lastname: Leopizzi
authors.affiliation: Dept. of Physiology and Pharmacology "V. Erspamer", Sapienza University of Roma, P.le Aldo Moro, 5, 00185 Roma, Italy. Electronic address: silvana.gaetani@uniroma1.it.
authors.forename: S
authors.initials: S
authors.lastname: Gaetani
authors.affiliation: Division of Anatomical Pathology, Dept. of Oncology, University Hospital of Siena, Via delle Scotte, 6, 53100 Siena, Italy. Electronic address: raffaellaguazzo@tiscali.it.
authors.forename: R
authors.initials: R
authors.lastname: Guazzo
authors.affiliation: Dept. of Experimental Medicine, Sapienza University of Roma, V.le Regina Elena, 324, 00161 Roma, Italy. Electronic address: sabrinabasciani@yahoo.it.
authors.forename: S
authors.initials: S
authors.lastname: Basciani
authors.affiliation: Dept. of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy. Electronic address: oreste.moreschini@uniroma1.it.
authors.forename: O
authors.initials: O
authors.lastname: Moreschini
authors.affiliation: Division of Anatomical Pathology, Dept. of Oncology, University Hospital of Siena, Via delle Scotte, 6, 53100 Siena, Italy. Electronic address: m.m.desanti@ao-siena.toscana.it.
authors.forename: M
authors.initials: M
authors.lastname: De Santi
authors.affiliation: Dept. of Biochemical Sciences, Sapienza University of Roma, P.le Aldo Moro, 5, 00185 Roma, Italy. Electronic address: roberto.scandurra@uniroma1.it.
authors.forename: R
authors.initials: R
authors.lastname: Scandurra
authors.affiliation: Dept. of Biochemical Sciences, Sapienza University of Roma, P.le Aldo Moro, 5, 00185 Roma, Italy. Electronic address: anna.scottodabusco@uniroma1.it.
authors.forename: A
authors.initials: A
authors.lastname: Scotto d'Abusco
mesh_terms: D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism*; D024043:Collagen Type II / Q000096:biosynthesis*; D004305:Dose-Response Relationship, Drug; D005109:Extracellular Matrix / Q000378:metabolism*; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D006025:Glycosaminoglycans / Q000096:biosynthesis*; D006801:Humans; D011509:Proteoglycans / Q000096:biosynthesis*
chemical_list: D024043:Collagen Type II; D006025:Glycosaminoglycans; D011509:Proteoglycans; D005944:Glucosamine
keywords: Collagen; Glucosamine; Glycosaminoglycan; Human primary chondrocytes; Micromasses; Peptidyl-derivative