title: Chitosan scaffold modified with D-(+) raffinose and enriched with thiol-modified gelatin for improved osteoblast adhesion.
abstract: The aim of the present study was to investigate whether chitosan-based scaffolds modified with D-(+) raffinose and enriched with thiol-modified gelatin could selectively improve osteoblast adhesion and proliferation. 2, 3 and 4.5% chitosan films were prepared. Chitosan suitability for tissue engineering was confirmed by protein adsorption assay. Scaffolds were incubated with a 2.5 mg ml(-1) BSA solution and the decrease of protein content in the supernatants was measured by spectrophotometry. Chitosan films were then enriched with thiol-modified gelatin and their ability to bind BSA was also measured. Then, 2% chitosan discs with or without thiol-modified gelatin were used as culture substrates for MC3T3-E1 cells. After 72 h cells were stained with trypan blue or with calcein AM and propidium iodide for morphology, viability and proliferation assays. Moreover, cell viability was measured at 48, 72, 96 and 168 h to obtain a growth curve. Chitosan films efficiently bound and retained BSA proportionally to the concentration of chitosan discs. The amount of protein retained was higher on chitosan enriched with thiol-modified gelatin. Moreover, chitosan discs allowed the adhesion and the viability of cells, but inhibited their proliferation. The functionalization of chitosan with thiol-modified gelatin enhanced cell spreading and proliferation. Our data confirm that chitosan is a suitable material for tissue engineering. Moreover, our data show that the enrichment of chitosan with thiol-modified gelatin enhances its biological properties.
authors.affiliation: Centro Universitario di Odontoiatria, University of Parma, Via Gramsci 14, 43126 Parma, Italy. IMEM-CNR National Research Council, Via Università 7, 43126 Parma, Italy. Dip. Scienze Biomediche, Biotecnologiche e Traslazionali, University of Parma, Via Gramsci 14, 43126 Parma, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Galli
authors.forename: L
authors.initials: L
authors.lastname: Parisi
authors.forename: L
authors.initials: L
authors.lastname: Elviri
authors.forename: A
authors.initials: A
authors.lastname: Bianchera
authors.forename: A
authors.initials: A
authors.lastname: Smerieri
authors.forename: P
authors.initials: P
authors.lastname: Lagonegro
authors.forename: S
authors.initials: S
authors.lastname: Lumetti
authors.forename: E
authors.initials: E
authors.lastname: Manfredi
authors.forename: R
authors.initials: R
authors.lastname: Bettini
authors.forename: G M
authors.initials: GM
authors.lastname: Macaluso
mesh_terms: D000818:Animals; D041702:BALB 3T3 Cells; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects / Q000502:physiology; D048271:Chitosan / Q000737:chemistry*; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005780:Gelatin / Q000737:chemistry*; D008422:Materials Testing; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D011887:Raffinose / Q000737:chemistry / Q000494:pharmacology*; D013438:Sulfhydryl Compounds / Q000737:chemistry; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods; D054457:Tissue Scaffolds
chemical_list: D020099:Coated Materials, Biocompatible; D013438:Sulfhydryl Compounds; D005780:Gelatin; D048271:Chitosan; D011887:Raffinose