title: The effects of different cross-linking conditions on collagen-based nanocomposite scaffolds-an in vitro evaluation using mesenchymal stem cells.
abstract: Nanocomposite scaffolds which aimed to imitate a bone extracellular matrix were prepared for bone surgery applications. The scaffolds consisted of polylactide electrospun nano/sub-micron fibres, a natural collagen matrix supplemented with sodium hyaluronate and natural calcium phosphate nano-particles (bioapatite). The mechanical properties of the scaffolds were improved by means of three different cross-linking agents: N-(3-dimethylamino propyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide in an ethanol solution (EDC/NHS/EtOH), EDC/NHS in a phosphate buffer saline solution (EDC/NHS/PBS) and genipin. The effect of the various cross-linking conditions on the pore size, structure and mechanical properties of the scaffolds were subsequently studied. In addition, the mass loss, the swelling ratio and the pH of the scaffolds were determined following their immersion in a cell culture medium. Furthermore, the metabolic activity of human mesenchymal stem cells (hMSCs) cultivated in scaffold infusions for 2 and 7 days was assessed. Finally, studies were conducted of cell adhesion, proliferation and penetration into the scaffolds. With regard to the structural stability of the tested scaffolds, it was determined that EDC/NHS/PBS and genipin formed the most effectively cross-linked materials. Moreover, it was discovered that the genipin cross-linked scaffold also provided the best conditions for hMSC cultivation. In addition, the infusions from all the scaffolds were found to be non-cytotoxic. Thus, the genipin and EDC/NHS/PBS cross-linked scaffolds can be considered to be promising biomaterials for further in vivo testing and bone surgery applications.
authors.affiliation: Department of Composites and Carbon Materials, Institute of Rock Structure and Mechanics, Academy of Sciences of the Czech Republic, V Holesovickach 41, Prague 8, 182 09, Czech Republic. Laboratory of Biomechanics, Department of Mechanics, Biomechanics and Mechatronics, Faculty of Mechanical Engineering, Czech Technical University in Prague, Technicka 4, Prague 6, 166 07, Czech Republic.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Suchý
authors.forename: Monika
authors.initials: M
authors.lastname: Šupová
authors.forename: Pavla
authors.initials: P
authors.lastname: Sauerová
authors.forename: Martina
authors.initials: M
authors.lastname: Verdánová
authors.forename: Zbyněk
authors.initials: Z
authors.lastname: Sucharda
authors.forename: Šárka
authors.initials: Š
authors.lastname: Rýglová
authors.forename: Margit
authors.initials: M
authors.lastname: Žaloudková
authors.forename: Radek
authors.initials: R
authors.lastname: Sedláček
authors.forename: Marie Hubálek
authors.initials: MH
authors.lastname: Kalbáčová
mesh_terms: D001672:Biocompatible Materials / Q000138:chemical synthesis; D001857:Bone Matrix / Q000737:chemistry; D018786:Bone Substitutes / Q000138:chemical synthesis*; D002448:Cell Adhesion / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D003094:Collagen / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D008422:Materials Testing; D045164:Mesenchymal Stem Cell Transplantation / Q000295:instrumentation* / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D053761:Nanocomposites / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D054457:Tissue Scaffolds
chemical_list: D001672:Biocompatible Materials; D018786:Bone Substitutes; D003432:Cross-Linking Reagents; D003094:Collagen