Every year in the UK, more than 300,000 hip, knee, shoulder, ankle or elbow devices are implanted into patients for the treatment of orthopaedic pain, disease and trauma. Secure fixation of these implants in bone is essential for the procedure's success, yet is challenging to achieve as bone is a living tissue that adapts and changes postoperatively. Researchers and industry strive to develop new technologies to improve fixation, with many aiming to take advantage of bone's living response by enabling it to grow into the implant. The design intent of these new technologies is always well-meaning, but to protect patients, it is necessary to pre-clinically test them, to confirm they are both safe and achieve their aim. However, there is a lack of appropriate methods for testing this. Traditional laboratory pre-clinical testing methods do not allow for testing with living bone samples and thus cannot measure implant bone ingrowth/adaptation. Live animal testing has ethical issues, is expensive and is complicated by anatomical differences and unknown loading. Computational models require input and validation data and so require a previous laboratory/animal/clinical study. The other alternative is clinical trial, which is effectively experimenting on patients. It also often requires years/decades of waiting to determine the outcome, and thus is only suitable as the final step of new product development. This research project aims to overcome limitations in pre-clinical testing by using a bioreactor system to enable implant fixation technologies to be tested against 'living' bone in the laboratory. The developed methods will be validated with established clinical technologies, before being applied to pre-clinically test a novel implant fixation concept. The long-term ambition for this research is to lower the risk for patients enrolling on clinical trials, reduce the need for ineffective live animal testing, and improve orthopaedic implants through enabling fixation technology to be optimised for in vivo performance.

Mass spectrometry, a technique that was initially applied to individual atoms and small molecules, is increasingly being used to look at much larger molecules such as proteins, not only individually, but also as large assemblies. These assemblies are often important molecular machines that control many different reactions in living cells. One of the problems with studying these assemblies is that they rarely exist in a single form, but rather are often found as mixtures of different numbers of proteins molecules that have been modified in some way. Most methods of analysis are therefore unable to study them since the results are effectively blurred by averaging across the different assemblies. Mass spectrometry of these large assemblies, however, is capable of separating the different components according to their mass to charge ratio, thereby allowing us to characterise by mass all the components within a mixed assembly. Until now mass spectrometry has not been able to tell us much about the shape of these assemblies. In order to do this we need to couple mass spectrometry to other methods to get three-dimensional images of these assemblies. This proposal therefore sets out to do this: to combine mass spectrometry with an established imaging technique known as electron microscopy. In addition, we have recently shown that we can measure the cross section of mixtures of large protein assemblies within a mass spectrometer. This has allowed us to deduce that their shape is close to what we would expect from analysis of crystals of the same assembly with X-rays, and we intend to develop this research further. We believe that together this combined has the potential to provide an incredibly powerful structural biology tool that will be able to tackle many protein assemblies that are currently intractable.

The flow response of 'soft materials' such as suspension, emulsions, (bio)gels and (bio)polymers, is of prime importance in a vast range of industries, e.g. foodstuffs and personal care products, and is the subject of intensive applied and fundamental research. Traditional characterization of these properties (viscosity, elasticity, creep, aging etc..) relies on rheological measurements, but it is now recognized that this alone is insufficient to fully understand, and thus optimize, the complex flow properties of soft materials. What is often required is characterization of their evolving microstructure during flow, allowing direct mapping of this evolution to their rheological response. We propose to develop a versatile module which will enable novel high quality imaging of micro-structure evolution and advanced velocimetry in rheometers in different geometries. The module will be complemented by a suite of analysis techniques. The combined capabilities will provide a new dimension in rheo-optical characterization and the module will bring these within reach of a variety of industrial and academic research laboratories.

This project addresses the application of high resolution and high sensitivity mass spectrometry to characterize the protein components in rodent scent marks. Recent research in rodent semiochemistry, in substantial part from the academic applicants laboratories, has revealed a depth and complexity to rodent chemical communication previously unanticipated. We are building a detailed picture of the receptor repertoire for these signals, and of the higher level processing that collates these signals into behavioural responses, but our understanding of the molecular composition of the scent marks is some way behind. Chemical communication is capable of conveying an incredibly subtle stream of information, especially between conspecifics. Scent marks, predominantly urinary, contain an astonishing array of information that transmits variable indicators of the state of the scent owner (e.g. health status, pregnancy, recent food ingested and time since deposition). This status information is primarily associated with proteins (lipocalins and ESPs) that provide information on genetically invariant parameters such as sex and individual identity. In this project, the student will bring to bear advanced mass spectrometric methodologies in the characterization of these proteins. The samples will be recovered from wild-caught rodents, and are sometimes vanishingly small (such as in tear secretions). The challenges in the study of these scent mark proteins are two fold. First, there is a pressing need for accurate quantification of the proteins in the scent mark. Second, it is clear that the highly polymorphic gene cluster that encodes these proteins is genetically unstable, leading to each wild animal being able to express a unique pattern of these proteins. Thus the challenges are both quantitative and qualitative. The student will address these challenges in collaboration with Waters, using a combination of intact mass profiling, making use of the high resolution QToF instruments and new algorithmic approaches to spectral deconvolution developed at Waters, label-free quantification using the Waters-developed MS^E analytical workflow, coupled with Hi3 peptide quantification, and through the use of selected reaction monitoring, using an artificial QconCAT concatenated standard peptide assembly (technology invented and patented by the academic partner). Finally, discovery of new polymorphic variants will be based on intact mass survey, followed by electron transfer dissociation (ETD, using a new front-end source designed at Waters but not yet widely available) to isolate and characterise the amino acid sequences of the variant proteins. The plan of the programme will follow the outline below, although we expect that from year 2 onwards, the student will take some responsibility for the direction of the research. Year 1: Induction, Design of QconCAT proteins, expression and validation, experience of wild rodent sample collection and diversity. Training in intact mass profiling, bioinformatics tools and peptide level label-fre and label-mediated quantification. Year 2: Development of ion mobility methodologies to improve resolution of complex mixtures of isoforms, and the use of gas phase cross-sectional area to assess the conformational stability and consequent degree of protonation on electrospray ionsation behaviour. Expression of recombinant lipocalins for model studies, appropriately engineered to alter electrostatic potential for charge state manipulation. Instruction on ETD fragmentation. Year 3: Application of ETD to discover amino acid sequence variation in new isoform variants, leading to quantification by surrogate peptides. Includes an exploration of the effect of sequence variation on ESI signal intensity and charge state for quantification. Year 4: Completion of thesis and papers, exploration of new areas of investigation.

Biomedical Materials have advanced dramatically over the last 50 years. Historically, they were considered as materials that formed the basis of a simple device, e.g. a hip joint or a wound dressing with a predominant tissue interface. However, biomedical materials have grown to now include the development of smart and responsive materials. Accordingly, such materials provide feedback regarding their changing physiological environment and are able to respond and adapt accordingly, for a range of healthcare applications. Two major areas underpinning this rapid development are advances in biomedical materials manufacture and their characterisation. Medical products arising from novel biomedical materials and the strategies to develop them are of great importance to the UK and Ireland. It is widely recognised that we have a rapidly growing and ageing population, with demand for more effective but also cost effective healthcare interventions, as identified in recent government White Paper and Foresight reports. This links directly to evidence of the world biomaterials market, estimated to be USD 70 billion (2016) and expected to grow to USD 149 billion by 2021 at a CAGR of 16%. To meet this demand an increase of 63% in biomedical materials engineering careers over the next decade is predicted. There is therefore a national need for a CDT to train an interdisciplinary cohort of students and provide them with a comprehensive set of skills so that they can compete in this rapidly growing field. In addition to the training of a highly skilled workforce, clinically and industrially led research will be performed that focuses on developing and translating smart and responsive biomaterials with a particular focus on higher throughput, greater reproducibility of manufacture and characterisation. We therefore propose a CDT in Advanced Biomedical Materials to address the need across The Universities of Manchester, Sheffield and The Centre for Research in Medical Devices (CÚRAM), Republic of Ireland (ROI). Our combined strength and track record in biomaterials innovation, translation and industrial engagement aligns the UK and ROI need with resource, skills, industrial collaboration and cohort training. This is underpinned strategically by the Biomedical Materials axis of the UK's £235 million investment of the Henry Royce Institute, led by Manchester and partner Sheffield. To identify key thematic areas of need the applicants led national Royce scoping workshops with 200 stakeholders through 2016 and 2017. Representation was from clinicians, industry and academia and a national landscape strategy was defined. From this we have defined priority research areas in bioelectronics, fibre technology, additive manufacturing and improved pre- clinical characterisation. In addition the need for improved manufacturing scale up and reproducibility was highlighted. Therefore, this CDT will have a focus on these specific areas, and training will provide a strongly linked multidisciplinary cohort of biomedical materials engineers to address these needs. All projects will have clinical, regulatory and industry engagement which will allow easy translation through our well established clinical trials units and positions the research well to interface with opportunities arising from 'Devolution Manchester', as Greater Manchester now controls long-term health and social care spending, ready for the full devolution of a budget of around £6 billion in 2016/17 which will continue through the CDT lifespan.