STUDIES ON KERATIN-BASED BIOMATERIALS FOR THE APPLICATION TO TISSUE ENGINEERING

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To use wool keratins in developing tissue engineering field, we have investigated the methods to reinforce the keratin materials by mixing with other biopolymers or chemical crosslinking etc., the modification of keratin materials with bioactive molecules, and the cell adhesion and proliferation on keratin materials. The following results were obtained.1.The composite keratin films with chitosan showed the antibacterial activity and the improved strength. In addition, chemical crosslinking of keratin with diepoxy compounds gave the highly flexible and water tolerant films.2.Cell adhesion peptide, RODS was linked to free cysteine residues of keratin. The surface coated with RGDS-carrying keratins showed the increased cell adhesion comparing with unmodified keratins.3.Lysozyme, as a model bioactive substance, was immobilized to free cysteine residues on freeze-dried keratin sponge. Lysozyme linked to sponge through S-C bond was stably maintained, while lysozyme linked through S-S bond was gradually released.4.The porous keratin sponges having homogenous pore structure with a mean pore diameter of approximately 100 μm were given by freezing the keratin solution at -20℃ for 3 days followed by the lyophilization. When mouse fibroblast cells were cultivated on the sponge, the maximum cell number per area on the sponge was 37 times higher than that on commercially available culture dish.5.To obtain a keratin film, which does not contain any additive but have high tenacity, the keratin powder prepared by spray-drying of reduced keratin solution was compression-molded. Homogenous transparent and highly strong plastic-like film was obtained by adding equal weight of 90% ethanol to keratin powder. The film had good water-tolerability and supported the cell adhesion and proliferation.