title: Real-time monitoring of peptide grafting onto chitosan films using capillary electrophoresis.
abstract: Chitosan, being antimicrobial and biocompatible, is attractive as a cell growth substrate. To improve cell attachment, arginine-glycine-aspartic acid-serine (RGDS) peptides were covalently grafted to chitosan films, through the widely used coupling agents 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC-HCl) and N-hydroxysuccinimide (NHS), via the carboxylic acid function of the RGDS molecule. The grafting reaction was monitored, for the first time, in real time using free-solution capillary electrophoresis (CE). This enabled fast separation and determination of the peptide and all other reactants in one separation with no sample preparation. Covalent RGDS peptide grafting onto the chitosan film surface was demonstrated using solid-state NMR of swollen films. CE indicated that oligomers of RGDS, not simply RGDS, were grafted on the film, with a likely hyperbranched structure. To assess the functional properties of the grafted films, cell growth was compared on control and peptide-grafted chitosan films. Light microscopy and polymerase chain reaction (PCR) analysis demonstrated greatly improved cell attachment to RGDS-grafted chitosan films.
authors: University of Western Sydney (UWS), Molecular Medicine Research Group (MMRG), Parramatta, 2150, Australia.
authors: Danielle L
authors: DL
authors: Taylor
authors: Joel J
authors: JJ
authors: Thevarajah
authors: Diksha K
authors: DK
authors: Narayan
authors: Patricia
authors: P
authors: Murphy
authors: Melissa M
authors: MM
authors: Mangala
authors: Seakcheng
authors: S
authors: Lim
authors: Richard
authors: R
authors: Wuhrer
authors: Catherine
authors: C
authors: Lefay
authors: Michael D
authors: MD
authors: O'Connor
authors: Marianne
authors: M
authors: Gaborieau
authors: Patrice
authors: P
authors: Castignolles
mesh_terms: D001672:Biocompatible Materials / Q000138:chemical synthesis* / Q000737:chemistry; D002448:Cell Adhesion; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D002477:Cells / Q000166:cytology; D048271:Chitosan / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D010455:Peptides / Q000737:chemistry*
chemical_list: D001672:Biocompatible Materials; D010455:Peptides; D048271:Chitosan