title: Preparation of Dual Functionalized Surfaces for Covalent Immobilization of BMP-6 and Adhesive Ligands for Biological Applications
creator: Martin, Volker
subject: 540
subject: 540 Chemistry and allied sciences
description: The median age of our population causes osteoporosis, bone fractures and disorders, which are also caused by multiple myeloma. In the past 25 years, regenerative medicine had gained in importance, especially for regeneration and renewal of bone tissue, which consists of different cell types composed in a very complex architecture. The growth factor bone morphogenetic protein 6 (BMP-6) belongs to the transforming growth factor β (TGF- β) superfamily and it induces the differentiation of mesenchymal stem cells into mature osteoblasts in bone leading to new bone formation. Besides induction of osteogenic differentiation, BMP-6 is also known to induce cell death in multiple myeloma cells in high concentrations. However, a systemic application is not practicable, since uncontrolled diffusion causes a wide range of side-effects. Immobilization of growth factors allows local treatment of bone fractures and defects, while it prevents uncontrolled release of growth factors. Furthermore, the required amount of growth factors can be reduced tremendously. The objective of this work was the covalent immobilization of BMP-6 co-presented with clicked integrin ligands on a structured gold nanoparticle (AuNP) platform, using blockcopolymer micellar nanolithography (BCMN) developed by Prof. Spatz and co-workers, to study integrin signaling in connection with growth factor responses. BMP-6 was selectively bound to gold nanoparticles organized in a hexagonal structure on the surface allowing to control the amount and density on the surface. I showed that surface co-presentation of BMP-6 and RGD or α5β1 integrin selective ligand promotes SMAD1/5 phosphorylation and osteogenic differentiation of the standard model system C2C12, even at amounts as low as 1 ng, whereas soluble BMP-6 application is significantly less effective. Additionally, BMP-6 was immobilized on gold nanostructured polyethylene glycol diacrylamide (PEG-DA) hydrogels containing different concentrations of cRGD in order to study the influence of the stiffness on the cell signaling. Furthermore, this approach was used to investigate the effect of immobilized BMP-6 in low doses on the multiple myeloma cell line OPM-2 to induce cell death.  This approach provides for the first time the successful presentation of BMP-6 in small and defined amounts on surfaces in combination with adhesive ligands. Furthermore, covalent immobilization hinders protein release while maintaining the biological activity of the growth factor.
date: 2017
type: Dissertation
type: info:eu-repo/semantics/doctoralThesis
type: NonPeerReviewed
format: application/pdf
identifier: https://archiv.ub.uni-heidelberg.de/volltextserverhttps://archiv.ub.uni-heidelberg.de/volltextserver/23489/1/Doktorarbeit%20Volker%20Martin.pdf
identifier: DOI:10.11588/heidok.00023489
identifier: urn:nbn:de:bsz:16-heidok-234893
identifier:   Martin, Volker  (2017) Preparation of Dual Functionalized Surfaces for Covalent Immobilization of BMP-6 and Adhesive Ligands for Biological Applications.  [Dissertation]     
relation: https://archiv.ub.uni-heidelberg.de/volltextserver/23489/
rights: info:eu-repo/semantics/openAccess
rights: http://archiv.ub.uni-heidelberg.de/volltextserver/help/license_urhg.html
language: eng