Investigation into the incorporation of RGD into polymers as a non-integrin selective strategy for tissue engineering

• Main Author: Perlin, Lynne
• Published: University of Sheffield 2008
• Subjects: 610.28
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.444266

This thesis describes the development of the 4-bromobenzylsulphonyl (4- Bbs) group as an enzymatically cleavable protecting group for the side chain of arginine. The protected arginine was utilised to synthesise RGD peptidcs both with and without a spacer arm and these peptides were incorporated into hydrogels. Hydrogels were made from 1,2-propandiol-3-methacrylate (glycerol methacrylate, GM MA) or butyl methacrylate (BMA) and 1,2-ethandiol dimethacrylate (ethylene glycol dimethacrylate, EGDMA) and photopolymerized as 60~m coatings. Removal of the 4-Bbs protecting group was achieved by incubation with the enzyme Glutathione-S-Transferase. Culture of human dermal fibroblasts on the materials showed significant improvements in cell adhesion and viability in serum free media on glycerol methacrylate hydrogels with nominal RGD concentrations of 1 ~mol/g or greater. A spacer arm between the peptide and the bulk was not necessary to promote cell attachment. No significant improvement in cell adhesion and viability to butyl methacrylate hydrogels was observed at any of the peptide concentrations tested. The effects of peptide concentration, GST pre-treatment of materials, cell passage number and culture with soluble RGD were investigated by examining cell morphology, adhesion, viability and F-actin organisation.