PhD Studentship: “GreenBone” - plant-derived matrices to improve bone regeneration
With the exception of blood, bone is the most widely transplanted material and it is estimated that more than 2.2 million bone graft operations are performed annually worldwide in orthopedics and dentistry to address bony defects. However, autologous bone grafting is associated with clinical drawbacks, such as limited tissue availability and increased patient morbidity and complications at the donor surgical site, which includes bacterial infection. The research goal of this project is to develop state-of-the-art biomaterials for bone transplantation that will stimulate rapid bone regeneration. The novel approach uses Plant-derived polysaccharides, mainly Rhamnogalacturonan-I pectins (RG-I), which provide the possibility to both enhance osteogenesis and limit inflammation at the surgical, graft site. RG-Is provide considerable flexibility in the delivery format being capable of being used as nanocoatings and applied as injectable gel based-scaffolds.
The objectives of the current proposal are to: Examine RG-I as a nanocoating at the surface of hydroxyapatite (HaP) bone substitute materials and develop RG-I gel-based graft materials to improve bone regeneration in patients with high risk of poor grafting outcomes.
This proposal is highly innovative aiming to hybridize hydroxyapatite (HaP) bone substitute materials (which have limited solubility and therefore release minimal concentrations of potentially osteogenic ions) with plant-derived (pectin RG-Is) nanocoatings to enhance their osteogenic capability in the early stage of bone healing. By exploiting the anti-inflammatory effects of RG-Is evidenced in preliminary studies there is possibility to drive wound healing thereby reducing surgical morbidity. The project will involve biochemical and in vitro studies. The methods used for biochemical part will include covalent coating of RG-I to a range of relevant surfaces and analyses of chemical and physical properties of the coatings with scanning electron microscopy, atomic force microscopy and confocal microscopy using immunofluorescence staining with specific RG-I’ antibodies. Bulk hydrogel RG-Is will be produced by adopting the technique described by Kracun SK (2015). Hydrogels will be characterised mechanically (tensile strength, elastic modulus, gelling time, water sorption profiles). The osteogenic and immunomodulatory properties of developed nanocoatings and gel-based RG-Is will be characterised with human primary cells and cell lines in terms of attachment (light, fluorescence and confocal microscopy), proliferation, cell viability, mineralization, cell cycle, expression of bone regeneration genes as well as markers of inflammation tested by techniques such as Real-Time PCR / ELISAs.
This project will generate several important outcomes including progression towards the development of pectin RG-I nanocoated bone substitute materials for enhanced bone regeneration.
S.K. Kracun, J. Schuckel, B. Westereng, L.G. Thygesen, R.N. Monrad, V.G. Eijsink, W.G. Willats, Biotechnology for biofuels, 8 (2015) 70.
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