title: Optimisation of UV irradiation as a binding site conserving method for crosslinking collagen-based scaffolds.
abstract: Short wavelength (λ = 254 nm) UV irradiation was evaluated over a range of intensities (0.06 to 0.96 J/cm(2)) as a means of cross-linking collagen- and gelatin-based scaffolds, to tailor their material characteristics whilst retaining biological functionality. Zero-link carbodiimide treatments are commonly applied to collagen-based materials, forming cross-links from carboxylate anions (for example the acidic E of GFOGER) that are an essential part of integrin binding sites on collagen. Cross-linking these amino acids therefore disrupts the bioactivity of collagen. In contrast, UV irradiation forms bonds from less important aromatic tyrosine and phenylalanine residues. We therefore hypothesised that UV cross-linking would not compromise collagen cell reactivity. Here, highly porous (~99 %) isotropic, collagen-based scaffolds were produced via ice-templating. A series of scaffolds (pore diameters ranging from 130-260 μm) with ascending stability in water was made from gelatin, two different sources of collagen I, or blends of these materials. Glucose, known to aid UV crosslinking of collagen, was added to some lower-stability formulations. These scaffolds were exposed to different doses of UV irradiation, and the scaffold morphology, dissolution stability in water, resistance to compression and cell reactivity was assessed. Stabilisation in aqueous media varied with both the nature of the collagen-based material employed and the UV intensity. Scaffolds made from the most stable materials showed the greatest stability after irradiation, although the levels of cross-linking in all cases were relatively low. Scaffolds made from pure collagen from the two different sources showed different optimum levels of irradiation, suggesting altered balance between stabilisation from cross-linking and destabilisation from denaturation. The introduction of glucose into the scaffold enhanced the efficacy of UV cross-linking. Finally, as hypothesized, cell attachment, spreading and proliferation on collagen materials were unaffected by UV cross-linking. UV irradiation may therefore be used to provide relatively low level cross-linking of collagen without loss of biological functionality.
authors.affiliation: Department of Materials Science and Metallurgy, University of Cambridge, 27 Charles Babbage Road, Cambridge, CB3 0FS, UK. nd313@cam.ac.uk.
authors.forename: Natalia
authors.initials: N
authors.lastname: Davidenko
authors.affiliation: Department of Materials Science and Metallurgy, University of Cambridge, 27 Charles Babbage Road, Cambridge, CB3 0FS, UK.
authors.forename: Daniel V
authors.initials: DV
authors.lastname: Bax
authors.affiliation: Department of Materials Science and Metallurgy, University of Cambridge, 27 Charles Babbage Road, Cambridge, CB3 0FS, UK.
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Schuster
authors.affiliation: Department of Biochemistry, University of Cambridge, Downing Site, Cambridge, CB2 1QW, UK.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Farndale
authors.affiliation: Department of Biochemistry, University of Cambridge, Downing Site, Cambridge, CB2 1QW, UK.
authors.forename: Samir W
authors.initials: SW
authors.lastname: Hamaia
authors.affiliation: Department of Materials Science and Metallurgy, University of Cambridge, 27 Charles Babbage Road, Cambridge, CB3 0FS, UK.
authors.forename: Serena M
authors.initials: SM
authors.lastname: Best
authors.affiliation: Department of Materials Science and Metallurgy, University of Cambridge, 27 Charles Babbage Road, Cambridge, CB3 0FS, UK.
authors.forename: Ruth E
authors.initials: RE
authors.lastname: Cameron
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D024042:Collagen Type I / Q000737:chemistry*; D006801:Humans; D008855:Microscopy, Electron, Scanning; D054457:Tissue Scaffolds; D014466:Ultraviolet Rays
chemical_list: D024042:Collagen Type I