Runx2 overexpression in bone marrow stromal cells accelerates bone formation in critically-sized femoral defects

The repair of large non-unions in long bones remains a significant clinical problem due to high failure rates and limited tissue availability for auto- and allografts. Many cell-based strategies for healing bone defects deliver bone marrow stromal cells to the defect site to take advantage of the inherent osteogenic capacity of this cell type. However, many factors, including donor age and ex vivo expansion of the cells, cause bone marrow stromal cells to lose their differentiation ability. To overcome these limitations, we have genetically engineered bone marrow stromal cells to constitutively overexpress the osteoblast specific transcription factor Runx2. In the present study, we examined Runx2-modified bone marrow stromal cells, delivered via poly(caprolactone) scaffolds loaded with type I collagen meshes, in critically-sized segmental defects in rats compared to unmodified cells, cell-free scaffolds and empty defects. Runx2 expression in bone marrow stromal cells accelerated healing of critically-sized defects compared to unmodified bone marrow stromal cells and defects receiving cell-free treatments. These findings provide an accelerated method for healing large bone defects which may reduce recovery time and the need for external fixation of critically-sized defects.

Elucidating the links between UV radiation and vitamin D synthesis : using an in vitro model

Ultraviolet radiation (UV) is the carcinogen that causes the most common malignancy in humans – skin cancer. However, moderate UV exposure is essential for producing vitaminDin our skin. VitaminDincreases the absorption of calcium from the diet, and adequate calcium is necessary for the building and maintenance of bones. Thus, low levels of vitamin D can cause osteomalacia and rickets and contribute to osteoporosis. Emerging evidence also suggests vitamin D may protect against falls, internal cancers, psychiatric conditions, autoimmune diseases and cardiovascular diseases. Since the dominant source of vitamin D is sunlight exposure, there is a need to understand what is a “balanced” level of sun exposure to maintain an adequate level of vitamin D but minimise the risks of eye damage, skin damage and skin cancer resulting from excessive UV exposure. There are many steps in the pathway from incoming solar UV to the eventual vitamin D status of humans (measured as 25-hydroxyvitamin D in the blood), and our knowledge about many of these steps is currently incomplete. This project begins by investigating the levels of UV available for synthesising vitamin D, and how these levels vary across seasons, latitudes and times of the day. The thesis then covers experiments conducted with an in vitro model, which was developed to study several aspects of vitamin D synthesis. Results from the model suggest the relationship between UV dose and vitamin D is not linear. This is an important input into public health messages regarding ‘safe’ UV exposure: larger doses of UV, beyond a certain limit, may not continue to produce vitamin D; however, they will increase the risk of skin cancers and eye damage. The model also showed that, when given identical doses of UV, the amount of vitamin D produced was impacted by temperature. In humans, a temperature-dependent reaction must occur in the top layers of human skin, prior to vitamin D entering the bloodstream. The hypothesis will be raised that cooler temperatures (occurring in winter and at high latitudes) may reduce vitamin D production in humans. Finally, the model has also been used to study the wavelengths of UV thought to be responsible for producing vitamin D. It appears that vitamin D production is limited to a small range of UV wavelengths, which may be narrower than previously thought. Together, these results suggest that further research is needed into the ability of humans to synthesise vitamin D from sunlight. In particular, more information is needed about the dose-response relationship in humans and to investigate the proposed impact of temperature. Having an accurate action spectrum will also be essential for measuring the available levels of vitamin D-effective UV. As this research continues, it will contribute to the scientific evidence-base needed for devising a public health message that will balance the risks of excessive UV exposure with maintaining adequate vitamin D.

Fit optimisation of a distal medial tibia plate

An iterative method for the fit optimisation of a pre-contoured fracture fixation plate for a given bone data set is presented. Both plate shape optimisation and plate fit quantification are conducted in a virtual environment utilising computer graphical methods and 3D bone and plate models. Two optimised shapes of the undersurface of an existing distal medial tibia plate were generated based on a dataset of 45 3D bone models reconstructed from computed tomography image data of Japanese tibiae. The existing plate shape achieved an anatomical fit on 13% of tibiae from the dataset. Modified plate 1 achieved an anatomical fit for 42% and modified plate 2 a fit for 67% of the bones. If either modified plate 1 or plate 2 is used, then the anatomical fit can be increased to 82% for the same dataset. Issues pertaining to any further improvement in plate fit/shape are discussed.

Engineering tubular bone constructs

Cell-sheet techniques have been proven effective in various soft tissue engineering applications. In this experiment, we investigated the feasibility of bone tissue engineering using a hybrid of mesenchymal stem cell (MSC) sheets and PLGA meshes. Porcine MSCs were cultured to a thin layer of cell sheets via osteogenic induction. Tube-like long bones were constructed by wrapping the cell sheet on to PLGA meshes resulting in constructs which could be cultured in spinner flasks, prior to implantation in nude rats. Our results showed that the sheets were composed of viable cells and dense matrix with a thickness of about 80–120 mm, mineral deposition was also observed in the sheet. In vitro cultures demonstrated calcified cartilage-like tissue formation and most PLGA meshes were absorbed during the 8-week culture period. In vivo experiments revealed that dense mineralized tissue was formed in subcutaneous sites and the 8- week plants shared similar micro-CT characteristics with native bone. The neo tissue demonstrated histological markers for both bone and cartilage, indicating that the bone formation pathway in constructs was akin to endochondral ossification, with the residues of PLGA having an effect on the neo tissue organization and formation. These results indicate that cell-sheet approaches in combination with custom-shaped scaffolds have potential in producing bone tissue.

Improved image contrast of the bone-muscle interface with 3T MRI compared to 1.5T MRI [Abstract]

Virtual 3D models of long bones are increasingly being used for implant design and research applications. The current gold standard for the acquisition of such data is Computed Tomography (CT) scanning. Due to radiation exposure, CT is generally limited to the imaging of clinical cases and cadaver specimens. Magnetic Resonance Imaging (MRI) does not involve ionising radiation and therefore can be used to image selected healthy human volunteers for research purposes. The feasibility of MRI as alternative to CT for the acquisition of morphological bone data of the lower extremity has been demonstrated in recent studies [1, 2]. Some of the current limitations of MRI are long scanning times and difficulties with image segmentation in certain anatomical regions due to poor contrast between bone and surrounding muscle tissues. Higher field strength scanners promise to offer faster imaging times or better image quality. In this study image quality at 1.5T is quantitatively compared to images acquired at 3T. --------- The femora of five human volunteers were scanned using 1.5T and 3T MRI scanners from the same manufacturer (Siemens) with similar imaging protocols. A 3D flash sequence was used with TE = 4.66 ms, flip angle = 15° and voxel size = 0.5 × 0.5 × 1 mm. PA-Matrix and body matrix coils were used to cover the lower limb and pelvis respectively. Signal to noise ratio (SNR) [3] and contrast to noise ratio (CNR) [3] of the axial images from the proximal, shaft and distal regions were used to assess the quality of images from the 1.5T and 3T scanners. The SNR was calculated for the muscle and bone-marrow in the axial images. The CNR was calculated for the muscle to cortex and cortex to bone marrow interfaces, respectively. --------- Preliminary results (one volunteer) show that the SNR of muscle for the shaft and distal regions was higher in 3T images (11.65 and 17.60) than 1.5T images (8.12 and 8.11). For the proximal region the SNR of muscles was higher in 1.5T images (7.52) than 3T images (6.78). The SNR of bone marrow was slightly higher in 1.5T images for both proximal and shaft regions, while it was lower in the distal region compared to 3T images. The CNR between muscle and bone of all three regions was higher in 3T images (4.14, 6.55 and 12.99) than in 1.5T images (2.49, 3.25 and 9.89). The CNR between bone-marrow and bone was slightly higher in 1.5T images (4.87, 12.89 and 10.07) compared to 3T images (3.74, 10.83 and 10.15). These results show that the 3T images generated higher contrast between bone and the muscle tissue than the 1.5T images. It is expected that this improvement of image contrast will significantly reduce the time required for the mainly manual segmentation of the MR images. Future work will focus on optimizing the 3T imaging protocol for reducing chemical shift and susceptibility artifacts.

Typical accuracy and quality control of a process for creating CT-Based virtual bone models

A pragmatic method for assessing the accuracy and precision of a given processing pipeline required for converting computed tomography (CT) image data of bones into representative three dimensional (3D) models of bone shapes is proposed. The method is based on coprocessing a control object with known geometry which enables the assessment of the quality of resulting 3D models. At three stages of the conversion process, distance measurements were obtained and statistically evaluated. For this study, 31 CT datasets were processed. The final 3D model of the control object contained an average deviation from reference values of −1.07±0.52 mm standard deviation (SD) for edge distances and −0.647±0.43 mm SD for parallel side distances of the control object. Coprocessing a reference object enables the assessment of the accuracy and precision of a given processing pipeline for creating CTbased 3D bone models and is suitable for detecting most systematic or human errors when processing a CT-scan. Typical errors have about the same size as the scan resolution.

Creation of a validated 3D finite element model of an ovine tibia

Introduction Ovine models are widely used in orthopaedic research. To better understand the impact of orthopaedic procedures computer simulations are necessary. 3D finite element (FE) models of bones allow implant designs to be investigated mechanically, thereby reducing mechanical testing. Hypothesis We present the development and validation of an ovine tibia FE model for use in the analysis of tibia fracture fixation plates. Material & Methods Mechanical testing of the tibia consisted of an offset 3-pt bend test with three repetitions of loading to 350N and return to 50N. Tri-axial stacked strain gauges were applied to the anterior and posterior surfaces of the bone and two rigid bodies – consisting of eight infrared active markers, were attached to the ends of the tibia. Positional measurements were taken with a FARO arm 3D digitiser. The FE model was constructed with both geometry and material properties derived from CT images of the bone. The elasticity-density relationship used for material property determination was validated separately using mechanical testing. This model was then transformed to the same coordinate system as the in vitro mechanical test and loads applied. Results Comparison between the mechanical testing and the FE model showed good correlation in surface strains (difference: anterior 2.3%, posterior 3.2%). Discussion & Conclusion This method of model creation provides a simple method for generating subject specific FE models from CT scans. The use of the CT data set for both the geometry and the material properties ensures a more accurate representation of the specific bone. This is reflected in the similarity of the surface strain results.

Attribute-based authenticated key exchange

We introduce the concept of attribute-based authenticated key exchange (AB-AKE) within the framework of ciphertext policy attribute-based systems. A notion of AKE-security for AB-AKE is presented based on the security models for group key exchange protocols and also taking into account the security requirements generally considered in the ciphertext policy attribute-based setting. We also extend the paradigm of hybrid encryption to the ciphertext policy attribute-based encryption schemes. A new primitive called encapsulation policy attribute-based key encapsulation mechanism (EP-AB-KEM) is introduced and a notion of chosen ciphertext security is de�ned for EP-AB-KEMs. We propose an EP-AB-KEM from an existing attribute-based encryption scheme and show that it achieves chosen ciphertext security in the generic group and random oracle models. We present a generic one-round AB-AKE protocol that satis�es our AKE-security notion. The protocol is generically constructed from any EP-AB-KEM that satis�es chosen ciphertext security. Instantiating the generic AB-AKE protocol with our EP-AB-KEM will result in a concrete one-round AB-AKE protocol also secure in the generic group and random oracle models.

Physiological investigation of periosteum structure and its application in periosteum tissue engineering

Although many different materials, techniques and methods, including artificial or engineered bone substitutes, have been used to repair various bone defects, the restoration of critical-sized bone defects caused by trauma, surgery or congenital malformation is still a great challenge to orthopedic surgeons. One important fact that has been neglected in the pursuit of resolutions for large bone defect healing is that most physiological bone defect healing needs the periosteum and stripping off the periosteum may result in non-union or non-healed bone defects. Periosteum plays very important roles not only in bone development but also in bone defect healing. The purpose of this project was to construct a functional periosteum in vitro using a single stem cell source and then test its ability to aid the repair of critical-sized bone defect in animal models. This project was designed with three separate but closely-linked parts which in the end led to four independent papers. The first part of this study investigated the structural and cellular features in periostea from diaphyseal and metaphyseal bone surfaces in rats of different ages or with osteoporosis. Histological and immunohistological methods were used in this part of the study. Results revealed that the structure and cell populations in periosteum are both age-related and site-specific. The diaphyseal periosteum showed age-related degeneration, whereas the metaphyseal periosteum is more destructive in older aged rats. The periosteum from osteoporotic bones differs from normal bones both in terms of structure and cell populations. This is especially evident in the cambial layer of the metaphyseal area. Bone resorption appears to be more active in the periosteum from osteoporotic bones, whereas bone formation activity is comparable between the osteoporotic and normal bone. The dysregulation of bone resorption and formation in the periosteum may also be the effect of the interaction between various neural pathways and the cell populations residing within it. One of the most important aspects in periosteum engineering is how to introduce new blood vessels into the engineered periosteum to help form vascularized bone tissues in bone defect areas. The second part of this study was designed to investigate the possibility of differentiating bone marrow stromal cells (BMSCs) into the endothelial cells and using them to construct vascularized periosteum. The endothelial cell differentiation of BMSCs was induced in pro-angiogenic media under both normoxia and CoCl2 (hypoxia-mimicking agent)-induced hypoxia conditions. The VEGF/PEDF expression pattern, endothelial cell specific marker expression, in vitro and in vivo vascularization ability of BMSCs cultured in different situations were assessed. Results revealed that BMSCs most likely cannot be differentiated into endothelial cells through the application of pro-angiogenic growth factors or by culturing under CoCl2-induced hypoxic conditions. However, they may be involved in angiogenesis as regulators under both normoxia and hypoxia conditions. Two major angiogenesis-related growth factors, VEGF (pro-angiogenic) and PEDF (anti-angiogenic) were found to have altered their expressions in accordance with the extracellular environment. BMSCs treated with the hypoxia-mimicking agent CoCl2 expressed more VEGF and less PEDF and enhanced the vascularization of subcutaneous implants in vivo. Based on the findings of the second part, the CoCl2 pre-treated BMSCs were used to construct periosteum, and the in vivo vascularization and osteogenesis of the constructed periosteum were assessed in the third part of this project. The findings of the third part revealed that BMSCs pre-treated with CoCl2 could enhance both ectopic and orthotopic osteogenesis of BMSCs-derived osteoblasts and vascularization at the early osteogenic stage, and the endothelial cells (HUVECs), which were used as positive control, were only capable of promoting osteogenesis after four-weeks. The subcutaneous area of the mouse is most likely inappropriate for assessing new bone formation on collagen scaffolds. This study demonstrated the potential application of CoCl2 pre-treated BMSCs in the tissue engineering not only for periosteum but also bone or other vascularized tissues. In summary, the structure and cell populations in periosteum are age-related, site-specific and closely linked with bone health status. BMSCs as a stem cell source for periosteum engineering are not endothelial cell progenitors but regulators, and CoCl2-treated BMSCs expressed more VEGF and less PEDF. These CoCl2-treated BMSCs enhanced both vascularization and osteogenesis in constructed periosteum transplanted in vivo.

A meta-study of SAP financials in the Queensland Government

This thesis consists of three related studies: an ERP Major Issues Study; an Historical Study of the Queensland Government Financial Management System; and a Meta-Study that integrates these and other related studies conducted under the umbrella of the Cooperative ERP Lifecycle Knowledge Management research program. This research provides a comprehensive view of ERP lifecycle issues encountered in SAP R/3 projects across the Queensland Government. This study follows a preliminary ERP issues study (Chang, 2002) conducted in five Queensland Government agencies. The Major Issues Study aims to achieve the following: (1) identify / explicate major issues in relation to the ES life-cycle in the public sector; (2) rank the importance of these issues; and, (3) highlight areas of consensus and dissent among stakeholder groups. To provide a rich context for this study, this thesis includes an historical recount of the Queensland Government Financial Management System (QGFMS). This recount tells of its inception as a centralised system; the selection of SAP and subsequent decentralisation; and, its eventual recentralisation under the Shared Services Initiative and CorpTech. This historical recount gives an insight into the conditions that affected the selection and ongoing management and support of QGFMS. This research forms part of a program entitled Cooperative ERP Lifecycle Knowledge Management. This thesis provides a concluding report for this research program by summarising related studies conducted in the Queensland Government SAP context: Chan (2003); Vayo et al (2002); Ng (2003); Timbrell et al (2001); Timbrell et al (2002); Chang (2002); Putra (1998); and, Niehus et al (1998). A study of Oracle in the United Arab Emirates by Dhaheri (2002) is also included. The thesis then integrates the findings from these studies in an overarching Meta-Study. The Meta-Study discusses key themes across all of these studies, creating an holistic report for the research program. Themes discussed in the meta-study include common issues found across the related studies; knowledge dynamics of the ERP lifecycle; ERP maintenance and support; and, the relationship between the key players in the ERP lifecycle.

An evaluation of IS-impact utility and intuitiveness

This study is conducted within the IS-Impact Research Track at Queensland University of Technology (QUT). The goal of the IS-Impact Track is, "to develop the most widely employed model for benchmarking information systems in organizations for the joint benefit of both research and practice" (Gable et al, 2006). IS-Impact is defined as "a measure at a point in time, of the stream of net benefits from the IS [Information System], to date and anticipated, as perceived by all key-user-groups" (Gable Sedera and Chan, 2008). Track efforts have yielded the bicameral IS-Impact measurement model; the "impact" half includes Organizational-Impact and Individual-Impact dimensions; the "quality" half includes System-Quality and Information-Quality dimensions. The IS-Impact model, by design, is intended to be robust, simple and generalisable, to yield results that are comparable across time, stakeholders, different systems and system contexts. The model and measurement approach employs perceptual measures and an instrument that is relevant to key stakeholder groups, thereby enabling the combination or comparison of stakeholder perspectives. Such a validated and widely accepted IS-Impact measurement model has both academic and practical value. It facilitates systematic operationalisation of a main dependent variable in research (IS-Impact), which can also serve as an important independent variable. For IS management practice it provides a means to benchmark and track the performance of information systems in use. From examination of the literature, the study proposes that IS-Impact is an Analytic Theory. Gregor (2006) defines Analytic Theory simply as theory that ‘says what is’, base theory that is foundational to all other types of theory. The overarching research question thus is "Does IS-Impact positively manifest the attributes of Analytic Theory?" In order to address this question, we must first answer the question "What are the attributes of Analytic Theory?" The study identifies the main attributes of analytic theory as: (1) Completeness, (2) Mutual Exclusivity, (3) Parsimony, (4) Appropriate Hierarchy, (5) Utility, and (6) Intuitiveness. The value of empirical research in Information Systems is often assessed along the two main dimensions - rigor and relevance. Those Analytic Theory attributes associated with the ‘rigor’ of the IS-Impact model; namely, completeness, mutual exclusivity, parsimony and appropriate hierarchy, have been addressed in prior research (e.g. Gable et al, 2008). Though common tests of rigor are widely accepted and relatively uniformly applied (particularly in relation to positivist, quantitative research), attention to relevance has seldom been given the same systematic attention. This study assumes a mainly practice perspective, and emphasises the methodical evaluation of the Analytic Theory ‘relevance’ attributes represented by the Utility and Intuitiveness of the IS-Impact model. Thus, related research questions are: "Is the IS-Impact model intuitive to practitioners?" and "Is the IS-Impact model useful to practitioners?" March and Smith (1995), identify four outputs of Design Science: constructs, models, methods and instantiations (Design Science research may involve one or more of these). IS-Impact can be viewed as a design science model, composed of Design Science constructs (the four IS-Impact dimensions and the two model halves), and instantiations in the form of management information (IS-Impact data organised and presented for management decision making). In addition to methodically evaluating the Utility and Intuitiveness of the IS-Impact model and its constituent constructs, the study aims to also evaluate the derived management information. Thus, further research questions are: "Is the IS-Impact derived management information intuitive to practitioners?" and "Is the IS-Impact derived management information useful to practitioners? The study employs a longitudinal design entailing three surveys over 4 years (the 1st involving secondary data) of the Oracle-Financials application at QUT, interspersed with focus groups involving senior financial managers. The study too entails a survey of Financials at four other Australian Universities. The three focus groups respectively emphasise: (1) the IS-Impact model, (2) the 2nd survey at QUT (descriptive), and (3) comparison across surveys within QUT, and between QUT and the group of Universities. Aligned with the track goal of producing IS-Impact scores that are highly comparable, the study also addresses the more specific utility-related questions, "Is IS-Impact derived management information a useful comparator across time?" and "Is IS-Impact derived management information a useful comparator across universities?" The main contribution of the study is evidence of the utility and intuitiveness of IS-Impact to practice, thereby further substantiating the practical value of the IS-Impact approach; and also thereby motivating continuing and further research on the validity of IS-Impact, and research employing the ISImpact constructs in descriptive, predictive and explanatory studies. The study also has value methodologically as an example of relatively rigorous attention to relevance. A further key contribution is the clarification and instantiation of the full set of analytic theory attributes.

Some applications of logic to feasibility in higher types

While it is commonly accepted that computability on a Turing machine in polynomial time represents a correct formalization of the notion of a feasibly computable function, there is no similar agreement on how to extend this notion on functionals, that is, what functionals should be considered feasible. One possible paradigm was introduced by Mehlhorn, who extended Cobham's definition of feasible functions to type 2 functionals. Subsequently, this class of functionals (with inessential changes of the definition) was studied by Townsend who calls this class POLY, and by Kapron and Cook who call the same class basic feasible functionals. Kapron and Cook gave an oracle Turing machine model characterisation of this class. In this article, we demonstrate that the class of basic feasible functionals has recursion theoretic properties which naturally generalise the corresponding properties of the class of feasible functions, thus giving further evidence that the notion of feasibility of functionals mentioned above is correctly chosen. We also improve the Kapron and Cook result on machine representation.Our proofs are based on essential applications of logic. We introduce a weak fragment of second order arithmetic with second order variables ranging over functions from NN which suitably characterises basic feasible functionals, and show that it is a useful tool for investigating the properties of basic feasible functionals. In particular, we provide an example how one can extract feasible programs from mathematical proofs that use nonfeasible functions.

Design and analysis of group key exchange protocols

A group key exchange (GKE) protocol allows a set of parties to agree upon a common secret session key over a public network. In this thesis, we focus on designing efficient GKE protocols using public key techniques and appropriately revising security models for GKE protocols. For the purpose of modelling and analysing the security of GKE protocols we apply the widely accepted computational complexity approach. The contributions of the thesis to the area of GKE protocols are manifold. We propose the first GKE protocol that requires only one round of communication and is proven secure in the standard model. Our protocol is generically constructed from a key encapsulation mechanism (KEM). We also suggest an efficient KEM from the literature, which satisfies the underlying security notion, to instantiate the generic protocol. We then concentrate on enhancing the security of one-round GKE protocols. A new model of security for forward secure GKE protocols is introduced and a generic one-round GKE protocol with forward security is then presented. The security of this protocol is also proven in the standard model. We also propose an efficient forward secure encryption scheme that can be used to instantiate the generic GKE protocol. Our next contributions are to the security models of GKE protocols. We observe that the analysis of GKE protocols has not been as extensive as that of two-party key exchange protocols. Particularly, the security attribute of key compromise impersonation (KCI) resilience has so far been ignored for GKE protocols. We model the security of GKE protocols addressing KCI attacks by both outsider and insider adversaries. We then show that a few existing protocols are not secure against KCI attacks. A new proof of security for an existing GKE protocol is given under the revised model assuming random oracles. Subsequently, we treat the security of GKE protocols in the universal composability (UC) framework. We present a new UC ideal functionality for GKE protocols capturing the security attribute of contributiveness. An existing protocol with minor revisions is then shown to realize our functionality in the random oracle model. Finally, we explore the possibility of constructing GKE protocols in the attribute-based setting. We introduce the concept of attribute-based group key exchange (AB-GKE). A security model for AB-GKE and a one-round AB-GKE protocol satisfying our security notion are presented. The protocol is generically constructed from a new cryptographic primitive called encapsulation policy attribute-based KEM (EP-AB-KEM), which we introduce in this thesis. We also present a new EP-AB-KEM with a proof of security assuming generic groups and random oracles. The EP-AB-KEM can be used to instantiate our generic AB-GKE protocol.