Untersuchungen zur Anbindung von Proteinen und Peptiden an Biomaterialien sowie zur Schädigung dieser Proteine und Peptide durch reaktive Sauerstoff-Spezies

摘要

Biomaterials in direct contact to body tissues may lead to inflammatory reactions with the possible formation of reactive molecule species. The extent of these reactions depends decisively on the property of the surface of the particular biomaterial which can be modified in a positive manner by means of the covalent coupling of physiologically active proteins and peptides. In this context the covalent coupling of proteins to biomaterial surfaces as well as the damage to proteins due to reactive molecule species was investigated: - the covalent coupling of different proteins and peptides to the surface of functionalised TiAl6V4 and Si3N4 as well as to the surface of polylactic acid plastic film grafted with polyacrylic acid was examined: at the surface of TiAl6V4 and Si3N4 about 70 molecules of horse radish peroxidase per 1 µm² could be covalently bound, whereas to the grafted polylactic acid plastic film about 25*105 molecules per 1 µm² could be bound. Experiments with further proteins show a correlation between the number of bound molecules and their size. - for the reactions of reactive molecule species with proteins 3-nitrotyrosine was chosen as a sensitive marker and an assay was established: after pre-column derivatisation with dimethylaminoazobenzenesulfonyl chloride (DABS-Cl) and subsequent analysis via High Performance Liquid Chromatography (HPLC) a detection limit in the lower pmol range could be achieved. For the quantification of even smaller amounts of 3-nitrotyrosine an enzyme linked immunosorbent assay (ELISA) with polyclonal antibodies was devel