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Title: The generation of 3D mineralised implants using human induced pluripotent stem cells for bone tissue engineering applications
Author: MacFarlane, Robert
ISNI:       0000 0004 7963 7548
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2019
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The management of critical size bone defects due to the effects of arthritis, arthroplasty surgery, or complex trauma is a challenging problem for orthopaedic surgeons. Traditional methods of autologous bone grafting or allografting, although often successful, are associated with a number of disadvantages and a range of complications, such as, limited tissue availability, donor site pain, lack of osteoinductive capacity, and variable clinical outcomes. Bone tissue engineering, using stem cells combined with a 3 dimensional (3D) delivery system to fill bone defects, is an exciting area of research with promising early results. This study takes this concept further with the production of novel 3D cellular based implants produced in hydrogels in a bioreactor using human induced pluripotent stem cells (hiPSCs). The aims of this project were to culture and differentiate hiPSCs towards and osteogenic lineage in vitro in both 2D and 3D environments, study the optimum culture conditions for osteogenesis, investigate various pharmacological agents for their ability to enhance osteogenesis, and to produce 3D mineralised cellular implants containing differentiated stem cells of an osteogenic lineage, and ultimately observe the ability of the implants to create novel bone tissue with a view to in vivo translation. hiPSCs were cultured in 2D tissue culture plates and differentiated using a 2-step integrated mesodermal and osteogenic induction strategy. The osteogenic effects of 2 statins, simvastatin and atorvastatin, and several commercially available anti-osteoporotic agents including strontium ranelate and teriparatide, were assessed for their comparative ability to enhance osteogenesis in hiPSCs. In a 3D culture system using calcium alginate bio-hydrogel encapsulation of hiPSCs, placed within a rotating wall bioreactor to achieve maximum cellular expansion, with pharmacological osteogenic induction/ enhancement using statins or teriparatide, 3D mineralised bone tissue constructs were subsequently produced. Tissue characterisation with histological staining, scanning electron microscopy (SEM), and tissue mineralisation and functionality, and osteogenic gene expression, was demonstrated. This study provides evidence for the use of hiPSCs as a patient-derived cell source with the potential for 3D osteogenic tissue production using pharmacological enhancement with statins or teriparatide, and the potential for clinical translation for patients with conditions characterised by bone loss.
Supervisor: Mantalaris, Athanasios Sponsor: Royal College of Surgeons of England
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral