title: Low molecular weight molecules of oyster nacre induce mineralization of the MC3T3-E1 cells.
abstract: The nacre layer from the pearl oyster shell is considered as a promising osteoinductive biomaterial. Nacre contains one or more signal molecules capable of stimulating bone formation. The identity and the mode of action of these molecules on the osteoblast differentiation were analyzed. Water-soluble molecules from nacre were fractionated according to dialysis, solvent extraction, and reversed-phase HPLC. The activity of a fraction composed of low molecular weight molecules in the mineralization of the MC3T3-E1 extracellular matrix was investigated. Mineralization of the preosteoblast cells was monitored according to alizarin red staining, Raman spectroscopy, scanning electron microscopy, and quantitative RT-PCR. Molecules isolated from nacre, ranging from 50 to 235 Da, induced a red alizarin staining of the preosteoblasts extracellular matrix after 16 days of culture. Raman spectroscopy demonstrated the presence of hydroxyapatite (HA) in samples treated with these molecules. Scanning electron microscopy pictures showed at the surface of the treated cells the occurrence of clusters of spherical particles resembling to HA. The treatment of cells with nacre molecules accelerated expression of collagen I and increased the mRNA expression of Runx2 and osteopontin. This study indicated that the nacre molecules efficient in bone cell differentiation are certainly different from proteins, and could be useful for in vivo bone repair.
authors.affiliation: Département des Milieux et Peuplements Aquatiques, UMR 5178, CNRS-MNHN Biologie des Organismes Marins et Ecosystèmes, ERT Valorisation de Molécules Bioactives d'Origine Marine, Paris, France. rousseam@gmx.net
authors.forename: Marthe
authors.initials: M
authors.lastname: Rousseau
authors.forename: Hélène
authors.initials: H
authors.lastname: Boulzaguet
authors.forename: Julie
authors.initials: J
authors.lastname: Biagianti
authors.forename: Denis
authors.initials: D
authors.lastname: Duplat
authors.forename: Christian
authors.initials: C
authors.lastname: Milet
authors.forename: Evelyne
authors.initials: E
authors.lastname: Lopez
authors.forename: Laurent
authors.initials: L
authors.lastname: Bédouet
mesh_terms: D000818:Animals; D002113:Calcification, Physiologic / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D024042:Collagen Type I / Q000096:biosynthesis; D045424:Complex Mixtures / Q000737:chemistry / Q000494:pharmacology*; D050659:Core Binding Factor Alpha 1 Subunit / Q000096:biosynthesis; D051379:Mice; D008970:Molecular Weight; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism*; D010012:Osteogenesis / Q000187:drug effects*; D053495:Osteopontin / Q000096:biosynthesis; D049911:Pinctada / Q000737:chemistry; D013997:Time Factors
chemical_list: D024042:Collagen Type I; D045424:Complex Mixtures; D050659:Core Binding Factor Alpha 1 Subunit; C493716:Runx2 protein, mouse; D053495:Osteopontin