D Stoppoloni1, L Politi2, M Leopizzi3, S Gaetani4, R Guazzo5, S Basciani6, O Moreschini7, M De Santi8, R Scandurra9, A Scotto d'Abusco10. 1. Dept. of Biochemical Sciences, Sapienza University of Roma, P.le Aldo Moro, 5, 00185 Roma, Italy. Electronic address: daniela.stoppoloni@uniroma1.it. 2. Dept. of Biochemical Sciences, Sapienza University of Roma, P.le Aldo Moro, 5, 00185 Roma, Italy. Electronic address: laura.politi@uniroma1.it. 3. 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. 4. 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. 5. Division of Anatomical Pathology, Dept. of Oncology, University Hospital of Siena, Via delle Scotte, 6, 53100 Siena, Italy. Electronic address: raffaellaguazzo@tiscali.it. 6. Dept. of Experimental Medicine, Sapienza University of Roma, V.le Regina Elena, 324, 00161 Roma, Italy. Electronic address: sabrinabasciani@yahoo.it. 7. Dept. of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy. Electronic address: oreste.moreschini@uniroma1.it. 8. 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. 9. Dept. of Biochemical Sciences, Sapienza University of Roma, P.le Aldo Moro, 5, 00185 Roma, Italy. Electronic address: roberto.scandurra@uniroma1.it. 10. Dept. of Biochemical Sciences, Sapienza University of Roma, P.le Aldo Moro, 5, 00185 Roma, Italy. Electronic address: anna.scottodabusco@uniroma1.it.
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.
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: Nuna Araújo; Carla S B Viegas; Eva Zubía; Joana Magalhães; Acácio Ramos; Maria M Carvalho; Henrique Cruz; João Paulo Sousa; Francisco J Blanco; Cees Vermeer; Dina C Simes Journal: Mar Drugs Date: 2020-12-07 Impact factor: 5.118
Authors: Alessia Mariano; Irene Bigioni; Roberto Mattioli; Antonella Di Sotto; Martina Leopizzi; Stefania Garzoli; Pier Francesco Mariani; Pietro Dalla Vedova; Sergio Ammendola; Anna Scotto d'Abusco Journal: Pharmaceuticals (Basel) Date: 2022-04-09
Authors: André Luiz A Pizzolatti; Florian Gaudig; Daniel Seitz; Carlos R M Roesler; Gean Vitor Salmoria Journal: Tissue Eng Regen Med Date: 2018-08-30 Impact factor: 4.169