title: Disulfide-crosslinked hyaluronan-gelatin hydrogel films: a covalent mimic of the extracellular matrix for in vitro cell growth.
abstract: A new disulfide crosslinking method was developed for the preparation of blended hyaluronan (HA)-gelatin hydrogels to form a synthetic, covalently linked mimic of the extracellular matrix (ECM). The HA and gelatin were chemically modified using 3,3'-dithiobis(propionic hydrazide) (DTP). After reduction with dithiothreitol (DTT), the thiol derivatives of HA (HA-DTPH) and gelatin (gelatin-DTPH) were obtained and characterized. To minimize interference with biological function, the degree of substitution of HA-DTPH and gelatin-DTPH was kept below 50%. Solutions of HA-DTPH and gelatin-DTPH in varying blends (20%, 40%, 60%, 80% gelatin) were prepared in 1% w/v NaCl and crosslinked by disulfide bond formation in air. Hydrogel films were dried and further crosslinked with dilute hydrogen peroxide. Disulfide crosslinked HA-DTPH, gelatin-DTPH, and blends thereof, were degradable enzymatically by collagenase and by hyaluronidase (HAse). The rapid digestion of the crosslinked 100% gelatin-DTPH film by collagenase was significantly retarded by the presence of 20% or 40% HA-DTPH. Addition of at least 40% w/v gelatin into the 100% HA-DTPH films significantly improved the attachment and spreading of Balb/c 3T3 murine fibroblasts seeded on the surface of the hydrogel. These results demonstrate that disulfide-crosslinked HA-gelatin hydrogels, a new type of covalent synthetic ECM, constitute biocompatible and biodegradable substrata for cell culture in vitro.
authors.affiliation: Department of Medicinal Chemistry, The University of Utah, 419 Wakara Way, Suite 205, Salt Lake City, 84108-1257, Utah, USA.
authors.forename: Xiao Zheng
authors.initials: XZ
authors.lastname: Shu
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Liu
authors.forename: Fabio
authors.initials: F
authors.lastname: Palumbo
authors.forename: Glenn D
authors.initials: GD
authors.lastname: Prestwich
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002448:Cell Adhesion; D002455:Cell Division; D002470:Cell Survival; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004220:Disulfides / Q000737:chemistry; D004229:Dithiothreitol / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005780:Gelatin / Q000737:chemistry*; D006820:Hyaluronic Acid / Q000737:chemistry*; D006821:Hyaluronoglucosaminidase / Q000737:chemistry; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D020100:Hydrogels; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D008756:Methylgalactosides / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008956:Models, Chemical; D011137:Polystyrenes / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry; D013438:Sulfhydryl Compounds; D013997:Time Factors
chemical_list: D001672:Biocompatible Materials; D003432:Cross-Linking Reagents; D004220:Disulfides; D020100:Hydrogels; D008756:Methylgalactosides; D011137:Polystyrenes; D013438:Sulfhydryl Compounds; C015233:gelatin film; C109801:hydrogel film; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D012965:Sodium Chloride; D005780:Gelatin; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase; D004229:Dithiothreitol