title: Fluorescence probe techniques to monitor protein adsorption-induced conformation changes on biodegradable polymers.
abstract: The study of protein adsorption and any associated conformational changes on interaction with biomaterials is of great importance in the area of implants and tissue constructs. This study aimed to evaluate some fluorescent techniques to probe protein conformation on a selection of biodegradable polymers currently under investigation for biomedical applications. Because of the fluorescence emanating from the polymers, the use of monitoring intrinsic protein fluorescence was precluded. A highly solvatochromic fluorescent dye, Nile red, and a well-known protein label, fluorescein isothiocyanate, were employed to study the adsorption of serum albumin to polycaprolactone and to some extent also to two starch-containing polymer blends (SPCL and SEVA-C). A variety of fluorescence techniques, steady state, time resolved, and imaging were employed. Nile red was found to leach from the protein, while fluorescein isothiocyanate proved useful in elucidating a conformational change in the protein and the observation of protein aggregates adsorbed to the polymer surface. These effects were seen by making use of the phenomenon of energy migration between the fluorescent tags to monitor interprobe distance and the use of fluorescence lifetime imaging to ascertain the surface packing of the protein on polymer.
authors.affiliation: 3B's Research Group-Biomaterials, Biodegradables and Biomimetics, Department of Polymer Engineering, University of Minho, 4710-057 Braga, Portugal. johan.benesch@dep.uminho.pt
authors.forename: Johan
authors.initials: J
authors.lastname: Benesch
authors.forename: Graham
authors.initials: G
authors.lastname: Hungerford
authors.forename: Klaus
authors.initials: K
authors.lastname: Suhling
authors.forename: Carolyn
authors.initials: C
authors.lastname: Tregidgo
authors.forename: João F
authors.initials: JF
authors.lastname: Mano
authors.forename: Rui L
authors.initials: RL
authors.lastname: Reis
mesh_terms: D020341:Absorbable Implants; D000327:Adsorption; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006801:Humans; D010078:Oxazines / Q000737:chemistry; D011091:Polyesters / Q000737:chemistry*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D012709:Serum Albumin / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013213:Starch / Q000737:chemistry
chemical_list: D010078:Oxazines; D011091:Polyesters; D011506:Proteins; D012709:Serum Albumin; C492325:starch polycaprolactone; C016240:polycaprolactone; D013213:Starch; D016650:Fluorescein-5-isothiocyanate; C044808:nile red