Design considerations of BMP-2 mimicking peptide immobilized scaffolds for use in bone tissue engineering
File(s)
Author(s)
Benge, Thomas
Type
Thesis
Abstract
Bone morphogenetic protein-2 (BMP-2) is prevalently researched and applied in the bone tissue engineering field due to its well characterised role in osteogenic differentiation and its ability to enhance bone regeneration in vivo. However, limitations, including insufficient local retention, high dosages used, recombinant growth factor immunogenicity, and recombinant growth factor cost, have prevented its widespread use as a bioactive addition to novel bone grafts in the clinic. This PhD thesis has explored solutions to these limitations by looking at the design and function of BMP-2 mimicking peptides as well as the design of novel methods for peptide immobilization quantification and immobilized BMP-2 mimicking peptides in 3D bone tissue engineering models.
Two, novel BMP-2 mimicking peptides (KTH1B & VAPP) were designed to recapitulate the anti-parallel beta sheets of the “knuckle” epitope’s secondary structure. These two, along with a literature “gold standard” original peptide (OP1), did not induce an osteogenic response when added as soluble additions to human mesenchymal stem cells (hMSCs) and osteoblastic cell lines. However, when surface coated at high concentration, they were detectable by BMP receptors with KTH1B and VAPP outperforming OP1. Additionally, experimental and modelled secondary structure analysis found that design mutations had meant that the desired beta sheet conformation had been lost, potentially contributing to lower binding and activity. To understand the potential of BMP-2 mimicking peptides immobilized, a novel immobilized peptide quantification assay was developed which can be used universally across multiple research fields and settings which was demonstrated through multiple applications including peptide immobilized surfaces, nanoclusters, and hydrogels. A hMSC encapsulated 3D hydrogel model with novel, orthogonal immobilized peptide incorporation was then designed to study immobilized BMP-2 mimicking peptide with...
Two, novel BMP-2 mimicking peptides (KTH1B & VAPP) were designed to recapitulate the anti-parallel beta sheets of the “knuckle” epitope’s secondary structure. These two, along with a literature “gold standard” original peptide (OP1), did not induce an osteogenic response when added as soluble additions to human mesenchymal stem cells (hMSCs) and osteoblastic cell lines. However, when surface coated at high concentration, they were detectable by BMP receptors with KTH1B and VAPP outperforming OP1. Additionally, experimental and modelled secondary structure analysis found that design mutations had meant that the desired beta sheet conformation had been lost, potentially contributing to lower binding and activity. To understand the potential of BMP-2 mimicking peptides immobilized, a novel immobilized peptide quantification assay was developed which can be used universally across multiple research fields and settings which was demonstrated through multiple applications including peptide immobilized surfaces, nanoclusters, and hydrogels. A hMSC encapsulated 3D hydrogel model with novel, orthogonal immobilized peptide incorporation was then designed to study immobilized BMP-2 mimicking peptide with...
Version
Open Access
Date Issued
2023-03-14
Date Awarded
01/09/2023
License URL
Advisor
Stevens, Molly
Jones, Julian
Wojciechowski, Jonathan
Sponsor
Engineering and Physical Sciences Research Council
Rosetrees Trust
Publisher Department
Materials
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)