Disulfide-crosslinked hyaluronan-gelatin hydrogel films: a covalent mimic of the extracellular matrix for in vitro cell growth.
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.
Shu
Xiao Zheng
XZ
Department of Medicinal Chemistry, The University of Utah, 419 Wakara Way, Suite 205, Salt Lake City, 84108-1257, Utah, USA.
Liu
Yanchun
Y
Palumbo
Fabio
F
Prestwich
Glenn D
GD
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
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