Sarcopenia, the loss of muscle mass and function as we age, affects all individuals from approximately the 4th decade of life and results in a poor quality of life. The mechanisms responsible for sarcopenia are unclear and it is likely that it is a multifactorial disease. However, it is hypothesised that an increase in systemic and/or muscle pro-inflammatory cytokine levels play a major role. Interferon induced protein 10 (IP10) is a chemokine that has been shown to be increased in serum as we age. Polyphenols are extracts from plants and have been shown to have anti-inflammatory effects with benefits in multiple diseases. Therefore, the main aim of this thesis was to establish an in vitro model to study the effect of increased levels of IP10 on muscle atrophy and inflammation and to examine whether resveratrol treatment was able to protect against IP10 induced effects on skeletal muscle atrophy or inflammation. Primary myoblasts were isolated from rat muscles and treated with 0.1µM, 1µM and 10µM of resveratrol to identify a functional concentration. Treatment of cells with 1µM of resveratrol increased myoblast growth, increased myotube diameter and decreased production of hydrogen peroxide of the cell media. Furthermore, treatment of myoblasts with 1µM resveratrol led to increases in the protein content of known resveratrol targets; MnSOD and catalase and caused transient increases in Sirt1 protein content. Treatment of cells with 1µM resveratrol also increased MnSOD and catalase content of myotubes and resulted in a decrease in the content of Sirt1, coupled with an increase in the acetylation status of proteins compared with untreated myotubes. As these data suggested that 1µM of resveratrol was functional in myotubes, this concentration was used to pre-treat myotubes before IP10 treatment. Treatment of myotubes with IP10 at levels found in older people (200pg/ml) led to decreases in myotube diameter and increases in the atrophy marker, Atrogin1. Pre-treatment of myotubes with resveratrol provided protection against both of these effects. Treatment of myotubes with a concentration of IP10 found in the young (150pg/ml) had no effect on markers of atrophy. Both concentrations of IP10 were found to have similar effects on cytokine release by myotubes and resveratrol treatment had only marginal effects on this. The second aim of this study was to identify an effective concentration of resveratrol in vivo and examine the effect of this on skeletal muscle force generation in adult and old mice. Treatment of mice with 125mg resveratrol/kg/day resulted in an increase in MnSOD and Sirt1 contents of skeletal muscle from young mice but had no effect on skeletal muscle force generation by EDL muscles of either adult or old mice. Overall data suggest that the increase in IP10 levels seen in ageing contributes to sarcopenia; however it is unlikely this acts through changes to the cytokine profile of muscle. Resveratrol prevented some atrophic effects induced by IP10, suggesting that, the mechanism through which resveratrol may protect against sarcopenia may be through the prevention of increases in atrophic pathways.

Introduction: Joint hypermobility syndrome (JHS) is a heritable connective tissue disorder associated with multiple joint laxity and pain. JHS is severe and disabling condition with a prevalence reaching 55% of patients attended physiotherapy with musculoskeletal symptoms. However, the literature is limited in quantity and quality to support the assessment and management strategies for people with JHS. Therefore, impairment, activity and participation were explored to identify the underlying problems. Methods: A cross-section design was employed to compare a group of adults with JHS against a matched control group. Neuromuscular impairments were explored through five domains: 1) pain intensity in the lower-limb joints was measured using visual analogue scales. 2) Achilles tendon stiffness was measured using the diagnostic ultrasound with strain-gauge myometer. 3) the plantar flexors strength was measured using the strain-gauge myometer. 4) knee proprioception was explored using the angle reproduction test. 5) gastrocnemius medius (GM) elasticity was quantified using the sonoelastography (SEG). Regarding the activity domain, both gait and vertical jump were analysed in terms of spatiotemporal, kinematics and kinetics using the Qualisys motion capture system, synchronised with the Kistler platform. The participation domain was assessed using the 12-item Short Form Health Survey (SF-12) and the Bristol Impact of Hypermobility questionnaire (BIoH). Additionally, the feasibility of the SEG was explored, and the intra-rater reliabilities for examining the Achilles tendon stiffness and gait kinematics were determined. Statistical Package for Social Sciences (SPSS) was used to conduct the statistical analysis. Results: The JHS group included 29 women and two men aged 38.52 ± 14.14 years (mean ± SD), while the control group included 29 women and two men aged 39.06 ± 12.43 years (mean ± SD). Various statistically significant differences were identified in the JHS group when compared to the control group, including increased pain intensity (all p ≤ 0.001), reduced Achilles tendon stiffness (p = 0.03), reduced plantar flexors strength (p = 0.01) and reduced non-dominant knee proprioception (p range 0.001 – 0.04). The gait and jump kinematics in the JHS group were mostly comparable to the control (p ≥ 0.05), with statistically significant reductions in moments (p ranged from 0.001 - 0.04) and power generation and absorption (p ranged from 0.001- 0.04) in the JHS group. Significant reductions in the participation level were evidenced in the JHS group, obtained from SF-12 (p ranged from 0.001 - 0.002), with significant impact from JHS (211.51 ± 39.28)/360 (mean ± standard deviation) obtained from the BIoH. Sonoelastography seems a feasible tool in terms of training, examination time, patient tolerance, and image analysis. High intra-rater reliability was demonstrated for examining the Achilles tendon stiffness (ICC ranged from 0.981 – 0.984), and moderate-high intra-rater reliability was demonstrated for examining gait kinematics (ICC ranged from 0.625 – 0.996). Conclusion: JHS has a multi-dimensional impact, causing neuromuscular impairment, activity limitations and participation restrictions. Assessment strategies should consider this multi-dimensional impact of the condition, and management strategies should be multi-disciplinary, aiming to target the problems identified in the current study. Key words: Joint hypermobility syndrome, impairment, activity, participation.

Tendons and ligaments play key roles in the musculoskeletal system in both man and animals. Both tissues can often be injured as result of contact based accidents or ageing and disease, causing discomfort, pain and increased susceptibility to degenerative joint disease. To date, tendon and ligament biology is relatively under-studied in healthy, non-diseased tissues. This information is essential to understand the pathology of these tissues and vital for future development of tendon and ligament tissue-engineered structures. This thesis aims to investigate the molecular and cellular differences between tendons and ligaments around the canine stifle joint. The biochemical composition, structural, and morphological characteristics were identified between the different regions of the intra- articular cranial cruciate ligament (CCL) and extra-articular medial collateral ligament (MCL), and the positional long digital extensor tendon (LDET) and energy storing superficial digital flexor tendons (SDFT). Differences in proteome composition were also assessed between CCL and LDET. Cells isolated from canine CCL and LDET were cultured in a 3D in vitro fibrin culture model and measured for differences in structural, biochemical and proteome composition. Statistical significant differences in extracellular matrix (ECM) composition in terms of glycosaminoglycan (GAG) and elastin content were primarily detected in CCL in comparison to the other three tissues. The CCL was also found to have morphological differences including less compact collagen architecture, differences in cell nuclei phenotype, and increased (GAG) and elastin content. Proteomic comparison between CCL and LDET resulted in significantly abundant fibrocartilage proteins such as collagen type II, aggrecan, versican and chondroadherin in CCL, while the LDET was more abundant in asporin and thrombospondin-4. 3D tendon and ligament constructs were able to recapitulate tendon and ligamentous tissue characteristics particularly with regards to ECM proteins present, however both construct were less abundant in ECM protein and contained a greater proportion of cellular proteins, corresponding with low collagen and high level of DNA content measured in both constructs. 3D tendon and ligament constructs derived from tendon and ligament cells had similar ECM, proteomic and structural composition, indicating that cell source may not be an important factor for tendon or ligament tissue engineering.

Abnormal hip joint morphology, associated with diseases such as femoroacetabular impingement (FAI) or developmental dysplasia of the hip (DDH), is thought to be a precursor of osteoarthritis (OA) in the hip. Changes in joint morphology alter the loading pattern through the hip, which results in damage to the tribological interface, including labral tears and/or labral- cartilage separation. Evidence shows that early intervention to repair the labrum is more beneficial than labral excision; however scientific understanding of the tissue and the effect of surgical treatments are limited. It is hypothesised that an in vitro natural hip simulation system could be used in biomechanical testing of hip joint tissues as well as generating labral damage which could be used to assess current and new surgical treatment methods for the labrum. Initial quantitative assays revealed human and porcine labral tissue to have higher collagen content but lower water and GAG content than articular cartilage. Histological staining identified the structure of collagen within the labrum and cartilage, as well as the dispersion of GAGs, in human and porcine tissue. Slight differences were seen between the two species with the human labrum containing more connective tissue compared to the porcine labrum, which was primarily composed of fibrocartilage, and less GAGs. Mechanical tests identified little variation between the compressive properties of the human and porcine labrum however, larger differences were identified in the tissues tensile properties, where by the human labrum was stronger than the porcine labrum. Labral tissue was also found to be weaker in compression in comparison to cartilage tissue. An in vitro natural hip model was successfully developed using clinically relevant conditions. The cup inclination angle was set at 45 °, a full ISO14242 gait cycle was applied to the joint with a peak load of 750 N, to account for porcine tissue. No labral or cartilage damage was observed after 10800 cycles. In vitro labral damage was also successfully developed, by increasing the acetabular cup angle to 60 ° and increasing the load by 50 %. The model was run through the full gait cycle for a minimum of 10800 cycles. Damage was classified using the Outerbridge and Lage systems. All types of labral damage outlined in the Lage classification system were identified within the model. Labral damage was found to progress from labral flattening, to radial fibrillation followed by longitudinal peripheral tears. The methodology and findings within this study can be used in future studies and can be advanced to mechanically test the soft tissues of the hip in situ, as well as the effect of labral damage on the functions of the hip joint and potential labral treatments.

No description available.

No description available.

This chapter aims to provide an overview of the dissertation presented here as a whole. It outlines the purpose, content and organisation of the following literature review (chapter 2) and empirical paper (chapter 3). This chapter also intends to show how the two main chapters fit together, and how they represent important research within the wider literature. Fibromyalgia (FM) is a chronic pain syndrome characterised by pain in the soft tissues of the body, general fatigue and sleep disturbance (Wolfe et al., 1990). Many individuals face a long and difficult journey to receive a diagnosis of FM. A recent survey reported that on average, respondents waited 2.3 years and saw 3.7 physicians before receiving a diagnosis of FM (Choy et al., 2010). Furthermore, individuals with FM have also reported that there is a considerable stigma associated with having the condition. In one qualitative study, women with FM reported feeling that others, including their physicians, questioned their credibility when reporting symptoms and their work ethic; and also implied that their illness was entirely psychological. As a result, the women reported that they coped with these difficulties either by withdrawing from social activity to avoid such experiences, or by putting on a façade that masked the true extent of their suffering (Asbring & Narvanen, 2002). Current medical and psychological treatments for FM are limited in success with regards to providing consistent benefits to the FM population as a whole (Abeles, Solitar, Pillinger & Abeles, 2008; Vlaeyen & Morley, 2005). Traditionally the dominant approach to intervention within both the medical and psychological fields is to focus on the reduction of negative symptoms. In contrast, the growing field of positive psychology continues to demonstrate the utility of exploring the processes and conditions that are conducive to optimal human functioning (Seligman & Csikszentmihalyi, 2000) as an alternative or complimentary approach to conventional methods of healthcare. Subjective well-being (SWB) can be defined as “a person’s cognitive and affective evaluations of his or her life” (Diener, Lucas & Oishi, 2002, p.63). A large body of evidence suggests that individuals who have higher SWB enjoy a range of positive outcomes, including health-related benefits (e.g. Deiner & Chan, 2011). The application of a positive psychology approach may be particularly relevant to FM, where there is growing evidence of a specific deficit in positive affect (PA), a major component of SWB. Chapter 2 of this thesis is a systematic review of the literature regarding PA in individuals with FM. It focuses on the quantitative literature and specifically aims to answer the question: is there a deficit in PA in individuals with Fibromyalgia relative to other pain conditions, general health conditions, and also the general population? The literature review begins by giving a rationale as to why exploring the evidence for a deficit in PA specific to the FM population is important. It also summarises the background literature regarding the structure of affect, as well as theories relating to the potential function, and proposed mechanism of action, of PA. Next, the method section outlines the systematic methods that were used to identify the relevant studies that are included in the review. This is followed by the results section, which succinctly presents a synthesis of the characteristics of the included studies, along with the key findings regarding PA. The discussion section considers how the findings answer the question of whether there is a specific deficit in PA within the FM population. It also considers the clinical implications of the findings. This is followed by an in-depth discussion of the potential limitations of the review, in terms of both the quality of the studies included and also the methodological considerations of the review process itself. Finally, recommendations for future research are made. It has been hypothesised that hope is a major contributor to well-being (Snyder, 2002). In non-clinical samples, the association between hope, particularly goal-focused hope, and SWB has been well documented (e.g. Snyder, 2002). More recently, mindfulness has also been identified as promoting increased SWB (e.g. Brown & Ryan, 2003). Chapter three of this thesis is an empirical paper that aims to add to the current literature by exploring the specific impact SWB has on improving FM-related symptoms and difficulties. It also builds on the existing literature in non-clinical samples by investigating if goal-focused hope and mindfulness significantly contribute to the promotion of SWB within the FM population. To achieve these aims, the research utilises structural equation modelling (SEM) techniques to simultaneously explore the relationships amongst the key study variables. This was done by pre-specifying a hypothesised model of how hope and mindfulness may lead to increased SWB in FM, based on past research. The extent to which this model fit the actual data collected was then examined. The empirical paper starts by considering the importance of SWB with regards to physical health outcomes. It also introduces the concepts of goal-focused hope and mindfulness, and begins to consider the theory behind how they may lead to higher SWB within the FM population. The method section then gives details of the study’s participants, measures and procedures. It also reports how the data was analysed, with a particular focus on a description of SEM. Next, the results section begins with details of how the data was prepared and includes findings from the preliminary analysis. The main focus of this section involves testing the hypothesised SEM model against the study data. Finally, the discussion section reflects on the study’s findings within the context of existing research and theory. Potential limitations of the study are considered, as well as the implications for future research and clinical practice. This section is concluded with a succinct summary of the study’s key contributions to the literature and how this should inform future work.

Patello-femoral problems affect nearly a quarter of the population and remain a common cause of knee replacement revision surgery. Minimally invasive treatments such as osteochondral substitutions are early interventions that can be used to prevent or delay the need for these replacements. Lack of pre-clinical testing is a major challenge in getting these promising treatments to clinical trials. The purpose of this research was to develop a platform that can get these products one step closer to clinical trials and hence getting them out in the market as a viable product for treating osteochondral lesions. The aim of this project was to develop and validate a design specification for the pre-clinical testing of natural patello-femoral joint (PFJ). A characterization study was carried out to investigate the suitable animal model required to simulate a human joint that requires osteochondral substitutions. The size of the porcine PFJ was closer to the human PFJ and the material properties of its cartilage were also similar to the human cartilage. Therefore, the porcine PFJ was chosen as the animal model to develop the methodologies for this project. A methodology was developed to investigate wear of the natural porcine PFJ by adapting a single station knee simulator to apply the porcine PFJ gait cycle to the joints. The position of the patella with respect to the femur determined through a contact point study was used to set up the samples in the simulator. A positive (cobalt chrome on natural cartilage) and negative (cartilage on cartilage) control was used to investigate the wear. This study showed the potential of using an Alicona Infinite focus G5 optical profiler to assess the change in cartilage topography in natural joints. The contact area and pressure in the PFJ was measured using Tekscan pressure sensors. This study showed the change in contact mechanics across a gait cycle and the effect of sample geometry on the contact mechanics of a joint. In-vitro simulation can reduce the need for animal testing and progress the preclinical trials for new tissue substitutions. Developing the methodology in a human knee is not practical. However, by establishing an animal model can bring this a step closer. The methods developed in this thesis can contribute towards creating a pre-clinical testing system that can be used to assess early interventions to the PFJ.

Aim: To develop protocols to digest staphylococcal enterotoxins B and C (SEB/SEC), toxic shock syndrome toxin 1 (TSST-1) and alpha haemolysin (AH), members of the pyrogenic toxin superantigen family (PTSAg). To develop a novel mass spectrometry method to analyse and compare urine samples from patients with rheumatoid arthritis (RA) and orthopaedic fracture patients for the presence of Staphylococcus aureus. Background: RA is a disease of unknown etiology; with a pathogenesis that is due to a mixture of genetic, immunological and environmental factors. A T-cell immune response to the presence of PTSAgs in the joints of RA patients has previously been described. A link has been proposed between pathogenic micro-organisms and the development of chronic, autoimmune conditions. Potential pathogenic mechanisms include the hygiene hypothesis and molecular mimicry. Due to the widespread prevalence of RA, it has been hypothesised that the pathogenesis could involve a common bacterium. In RA, one potential bacterial candidate that has been suggested is S.aureus. Current published data averages the presence of S.aureus in the general population at 30% (nasal/nasopharyngeal swabs). However, our unpublished data suggests immune complexes containing S.aureus antigens are detectable in urine. xii Methods: Mid-stream urine samples were collected from the rheumatology and orthopaedic departments of the Royal Lancaster Infirmary (RLI), UK. Urine samples were analysed by western blot and mass spectrometry. Results: 56.4% of RA patients showed the presence of at least one staphylococcal toxin in their urine compared with 27.1% of fracture patients. Conclusion: Our work demonstrates an increased presence of bacterial toxins in urine from RA patients, compared to the fracture controls and the current literature. This study is the first to demonstrate the presence of common staphylococcal enterotoxins in RA patient urine, raising the question of what role they may have in the disease pathogenesis, given that these patients have no active infections. This raises questions of whether the bacteria and their toxins are involved in an individual’s likelihood of getting RA; are those people with RA more likely to have S.aureus infections due to their immunological state? The presence of S.aureus in RA patient tissues warrants further investigation to determine if it is causative of, or a result of RA diagnosis.

No description available.