Development of zonal cartilage constructs : effects of chondrocyte subpopulation, compressive stimulation, and culture models

Articular cartilage is a highly resilient tissue located at the ends of long bones. It has a zonal structure, which has functional significance in load-bearing. Cartilage does not spontaneously heal itself when damaged, and untreated cartilage lesions or age-related wear often lead to osteoarthritis (OA). OA is a degenerative condition that is highly prevalent, age-associated, and significantly affects patient mobility and quality of life. There is no cure for OA, and patients usually resort to replacing the biological joint with an artificial prosthesis. An alternative approach is to dynamically regenerate damaged or diseased cartilage through cartilage tissue engineering, where cells, materials, and stimuli are combined to form new cartilage. However, despite extensive research, major limitations remain that have prevented the wide-spread application of tissue-engineered cartilage. Critically, there is a dearth of information on whether autologous chondrocytes obtained from OA patients can be used to successfully generate cartilage tissues with structural hierarchy typically found in normal articular cartilage. I aim to address these limitations in this thesis by showing that chondrocyte subpopulations isolated from macroscopically normal areas of the cartilage can be used to engineer stratified cartilage tissues and that compressive loading plays an important role in zone-dependent biosynthesis of these chondrocytes. I first demonstrate that chondrocyte subpopulations from the superficial (S) and middle/deep (MD) zones of OA cartilage are responsive to compressive stimulation in vitro, and that the effect of compression on construct quality is zone-dependent. I also show that compressive stimulation can influence pericelluar matrix production, matrix metalloproteinase secretion, and cytokine expression in zonal chondrocytes in an alginate hydrogel model. Subsequently, I focus on recreating the zonal structure by forming layered constructs using the alginate-released chondrocyte (ARC) method either with or without polymeric scaffolds. Resulting zonal ARC constructs had hyaline morphology, and expressed cartilage matrix molecules such as proteoglycans and collagen type II in both scaffold-free and scaffold-based approaches. Overall, my findings demonstrate that chondrocyte subpopulations obtained from OA joints respond sensitively to compressive stimulation, and are able to form cartilaginous constructs with stratified organization similar to native cartilage using the scaffold-free and scaffold-based ARC technique. The ultimate goal in tissue engineering is to help provide improved treatment options for patients suffering from debilitating conditions such as OA. Further investigations in developing functional cartilage replacement tissues using autologous chondrocytes will bring us a step closer to improving the quality of life for millions of OA patients worldwide.

Application of a human bone engineering platform to an in vitro and in vivo breast cancer metastasis model

Breast cancer in its advanced stage has a high predilection to the skeleton. Currently, treatment options of breast cancer-related bone metastasis are restricted to only palliative therapeutic modalities. This is due to the fact that mechanisms regarding the breast cancer celI-bone colonisation as well as the interactions of breast cancer cells with the bone microenvironment are not fully understood, yet. This might be explained through a lack of appropriate in vitro and in vivo models that are currently addressing the above mentioned issue. Hence the hypothesis that the translation of a bone tissue engineering platform could lead to improved and more physiological in vitro and in vivo model systems in order to investigate breast cancer related bone colonisation was embraced in this PhD thesis. Therefore the first objective was to develop an in vitro model system that mimics human mineralised bone matrix to the highest possible extent to examine the specific biological question, how the human bone matrix influences breast cancer cell behaviour. Thus, primary human osteoblasts were isolated from human bone and cultured under osteogenic conditions. Upon ammonium hydroxide treatment, a cell-free intact mineralised human bone matrix was left behind. Analyses revealed a similar protein and mineral composition of the decellularised osteoblast matrix to human bone. Seeding of a panel of breast cancer cells onto the bone mimicking matrix as well as reference substrates like standard tissue culture plastic and collagen coated tissue culture plastic revealed substrate specific differences of cellular behaviour. Analyses of attachment, alignment, migration, proliferation, invasion, as well as downstream signalling pathways showed that these cellular properties were influenced through the osteoblast matrix. The second objective of this PhD project was the development of a human ectopic bone model in NOD/SCID mice using medical grade polycaprolactone tricalcium phosphate (mPCL-TCP) scaffold. Human osteoblasts and mesenchymal stem cells were seeded onto an mPCL-TCP scaffold, fabricated using a fused deposition modelling technique. After subcutaneous implantation in conjunction with the bone morphogenetic protein 7, limited bone formation was observed due to the mechanical properties of the applied scaffold and restricted integration into the soft tissue of flank of NOD/SCID mice. Thus, a different scaffold fabrication technique was chosen using the same polymer. Electrospun tubular scaffolds were seeded with human osteoblasts, as they showed previously the highest amount of bone formation and implanted into the flanks of NOD/SCID mice. Ectopic bone formation with sufficient vascularisation could be observed. After implantation of breast cancer cells using a polyethylene glycol hydrogel in close proximity to the newly formed bone, macroscopic communication between the newly formed bone and the tumour could be observed. Taken together, this PhD project showed that bone tissue engineering platforms could be used to develop an in vitro and in vivo model system to study cancer cell colonisation in the bone microenvironment.

Waterfall : Nature and Culture

Many people take pleasure in visiting waterfalls and much has been written on the subject. Numerous accounts of Niagara Falls were published after Hennepin's late seventeenth-centure descriptions, particularly from the early nineteenth century, but is was only later that other waterfalls became the subject of books. George Holley's Niagara and Other Famous Cataracts of the World, published in 1883, and John Gibson's Great Waterfalls, Cataracts and Geysers, published in 1887, are early examples of global accounts of major falls. Most books about waterfalls are guides to the falls of a particular country, state or region. Apart from a few slim illustrated volumes, few books have been puslished on the world's waterfalls since Edward Rashleigh's Among the Waterfalls (1935). Most of these are slim pictorial volumes, some aimed at the children's market. Geologist Richard Maxwell Pearl published a series of waterfall articles in his journal Earth Science between 1973 and 1975, apparently with the intention of turning them into a book, but this never materialized. My book, the culmination of more than a decade of waterfalls research, is comprehensive in its approach, but is not intended to describe as many of the world's waterfalls as possible. This is far from my aim, and readers may be disappointed at my omission of falls they feel deserved mention. What I have attempted to do is celebrate the delights of these beautiful wonders of nature by considering them from many points of view, emphasizing the roles that they play in the human experience. To be as representative as possible, I draw on examples of waterfalls from all over the world, some famous, many not. North and South America, Europe, Africa, Asia and Oceania and, with recent global warming, the Earth's polar regions, all feature in the discussion. Even though there are already enough books and articles about Niagara Falls to fill a large library, it has been impossible to avoid making frequent reference to this great cataract, which has been so important in the history of travel and tourism, power generation, urban development and art. Amoung the issues that I consider is the human impact on waterfalls, particularly the effects of hydropower schemes and tourism development. Also considered are artificial waterfalls, which have long been features of the designed landscape. Their contemporay role is poignantly exemplified in the design of the National September 11 Memorial, in which the footprints of the Twin Towers are traced by walls of waterfalls. A geographer and urban and regional planner by training, I have ventured into many other fields of knowledge that are outside my areas of expertise. I apologize for any errors that I may have made in my book and invite correction.

Data mining traditional Chinese medicine (TCM) lessons learnt from mining in law and allopathic medicine

Key decisions at the collection, pre-processing, transformation, mining and interpretation phase of any knowledge discovery from database (KDD) process depend heavily on assumptions and theorectical perspectives relating to the type of task to be performed and characteristics of data sourced. In this article, we compare and contrast theoretical perspectives and assumptions taken in data mining exercises in the legal domain with those adopted in data mining in TCM and allopathic medicine. The juxtaposition results in insights for the application of KDD for Traditional Chinese Medicine.

Experimental evaluation of the culture performance of three exotic and a ‘local’ Giant Freshwater Prawn (Macrobrachium rosenbergii, De Man 1879) culture strain in Fiji

Giant freshwater prawn (GFP; Macrobrachium rosenbergii) aquaculture has expanded rapidly since 1990. Most local culture industries, however, have developed in an unsystematic way. Fiji has a small culture industry producing the ‘Anuenue’ strain; however, performance of this strain has never been systematically evaluated. Recently, some Fijian farmers have reported declines in stock productivity. The current project evaluated the relative performance of three exotic strains with different genetic backgrounds from Malaysia, Indonesia and Vietnam, against the ‘local’ strain in Fiji in a 4 × 3 replicated pond trial experiment. A total of 5827 prawns were harvested after 143 days growout. Individual growth rate and relative survival of the Fiji strain were not statistically different from any of the introduced strains, but Vietnam strain was superior to that of the Malaysia strain. Genetic diversity showed significant differences in variability among strains, with the Malaysian strain displaying the lowest genetic diversity. Indonesia strain showed that females were reaching maturation earlier than other strains and were smaller in size. This study suggests that Malaysian and Indonesian strains would constitute a poor choice for Fiji, whereas the Vietnam strain consistently performed well on all criteria measured. High variation among replicate ponds within strains unfortunately confounded among-strain variation.

Analyzing the literature for the link between the conservative Islamic culture of Saudi Arabia and the design of sustainable housing

Saudi Arabia experiences housing shortage for mid and low-income families, which is caused by rapid population growth. This condition is worsened by the fact that the current housing supply has problems in meeting both sustainable requirements and cultural needs of those families. This paper aims to investigate the link between the unique conservative Saudi culture and the design of sustainable housing, while keeping the housing cost affordable for mid and low-income families. The paper is based on a review of literatures on the issues of the Islamic culture and how can they be integrated into the design process of a Saudi house. Findings from literature reveiw suggest several design requirements for accommodating the conservative Saudi Culture in low cost sustainable houses. Such requirements include the implementation of proper usage of windows, and house orientation with a courtyard inside rather than facing the main street will provide natural ventilation while maintaining privacy. The main contribution to the body of knowledge is that this is a new approach to sustainable housing in Saudi Arabia considering not only energy use and architectural design issues but also socio-cultural issues as an essential part of sustainability.

Fast and robust stereo matching algorithms for mining automation

The mining environment, being complex, irregular, and time-varying, presents a challenging prospect for stereo vision. For this application, speed, reliability, and the ability to produce a dense depth map are of foremost importance. This paper evaluates a number of matching techniques for possible use in a stereo vision sensor for mining automation applications. Area-based techniques have been investigated because they have the potential to yield dense maps, are amenable to fast hardware implementation, and are suited to textured scenes. In addition, two nonparametric transforms, namely, rank and census, have been investigated. Matching algorithms using these transforms were found to have a number of clear advantages, including reliability in the presence of radiometric distortion, low computational complexity, and amenability to hardware implementation.

Fast and robust stereo matching algorithms for mining automation

The mining environment, being complex, irregular and time varying, presents a challenging prospect for stereo vision. The objective is to produce a stereo vision sensor suited to close-range scenes consisting primarily of rocks. This sensor should be able to produce a dense depth map within real-time constraints. Speed and robustness are of foremost importance for this investigation. A number of area based matching metrics have been implemented, including the SAD, SSD, NCC, and their zero-meaned versions. The NCC and the zero meaned SAD and SSD were found to produce the disparity maps with the highest proportion of valid matches. The plain SAD and SSD were the least computationally expensive, due to all their operations taking place in integer arithmetic, however, they were extremely sensitive to radiometric distortion. Non-parametric techniques for matching, in particular, the rank and the census transform, have also been investigated. The rank and census transforms were found to be robust with respect to radiometric distortion, as well as being able to produce disparity maps with a high proportion of valid matches. An additional advantage of both the rank and the census transform is their amenability to fast hardware implementation.

Suitability of non-parametric stereo matching algorithms for mining automation

The mining environment, being complex, irregular and time varying, presents a challenging prospect for stereo vision. For this application, speed, reliability, and the ability to produce a dense depth map are of foremost importance. This paper assesses the suitability of a number of matching techniques for use in a stereo vision sensor for close range scenes consisting primarily of rocks. These include traditional area-based matching metrics, and non-parametric transforms, in particular, the rank and census transforms. Experimental results show that the rank and census transforms exhibit a number of clear advantages over area-based matching metrics, including their low computational complexity, and robustness to certain types of distortion.

Fast and robust stereo matching algorithms for mining automation

The mining environment, being complex, irregular and time varying, presents a challenging prospect for stereo vision. For this application, speed, reliability, and the ability to produce a dense depth map are of foremost importance. This paper evaluates a number of matching techniques for possible use in a stereo vision sensor for mining automation applications. Area-based techniques have been investigated because they have the potential to yield dense maps, are amenable to fast hardware implementation, and are suited to textured scenes. In addition, two non-parametric transforms, namely, the rank and census, have been investigated. Matching algorithms using these transforms were found to have a number of clear advantages, including reliability in the presence of radiometric distortion, low computational complexity, and amenability to hardware implementation.

Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes

Regenerative medicine-based approaches for the repair of damaged cartilage rely on the ability to propagate cells while promoting their chondrogenic potential. Thus, conditions for cell expansion should be optimized through careful environmental control. Appropriate oxygen tension and cell expansion substrates and controllable bioreactor systems are probably critical for expansion and subsequent tissue formation during chondrogenic differentiation. We therefore evaluated the effects of oxygen and microcarrier culture on the expansion and subsequent differentiation of human osteoarthritic chondrocytes. Freshly isolated chondrocytes were expanded on tissue culture plastic or CultiSpher-G microcarriers under hypoxic or normoxic conditions (5% or 20% oxygen partial pressure, respectively) followed by cell phenotype analysis with flow cytometry. Cells were redifferentiated in micromass pellet cultures over 4 weeks, under either hypoxia or normoxia. Chondrocytes cultured on tissue culture plastic proliferated faster, expressed higher levels of cell surface markers CD44 and CD105 and demonstrated stronger staining for proteoglycans and collagen type II in pellet cultures compared with microcarrier-cultivated cells. Pellet wet weight, glycosaminoglycan content and expression of chondrogenic genes were significantly increased in cells differentiated under hypoxia. Hypoxia-inducible factor-3alpha mRNA was up-regulated in these cultures in response to low oxygen tension. These data confirm the beneficial influence of reduced oxygen on ex vivo chondrogenesis. However, hypoxia during cell expansion and microcarrier bioreactor culture does not enhance intrinsic chondrogenic potential. Further improvements in cell culture conditions are therefore required before chondrocytes from osteoarthritic and aged patients can become a useful cell source for cartilage regeneration.

Engineering across cultures : new learning resources for intercultural competency in engineering

BACKGROUND The work described in this paper has emerged from an ALTC/OLT funded project, Exploring Intercultural Competency in Engineering. The project indentified many facets of culture and intercultural competence that go beyond a culture-as-nationality paradigm. It was clear from this work that resources were needed to help engineering educators introduce students to the complex issues of culture as they relate to engineering practice. A set of learning modules focussing on intercultural competence in engineering practice were developed early on in the project. Through the OLT project, these modules have been expanded into a range of resources covering various aspects of culture in engineering. Supporting the resources, an eBook detailing the ins and outs of intercultural competency has also been developed to assist engineering educators to embed opportunities for students to develop skills in unpacking and managing cross-cultural challenges in engineering practice. PURPOSE This paper describes the key principles behind the development of the learning modules, the areas they cover and the eBook developed to support the modules. The paper is intended as an introduction to the approaches and resources and extends an invitation to the community to draw from, and contribute to this initial work. DESIGN/METHOD A key aim of this project was to go beyond the culture-as-nationality approach adopted in much of the work around intercultural competency (Deardorff, 2011). The eBook explores different dimensions of culture such as workplace culture, culture’s influence on engineering design, and culture in the classroom. The authors describe how these connect to industry practice and explore what they mean for engineering education. The packaged learning modules described here have been developed as a matrix of approaches moving from familiar known methods through complicated activities relying to some extent on expert knowledge. Some modules draw on the concept of ‘complex un-order’ as described in the ‘Cynefin domains’ proposed by Kurtz and Snowden (2003). RESULTS Several of the modules included in the eBook have already been trialled at a variety of institutions. Feedback from staff has been reassuringly positive so far. Further trials are planned for second semester 2012, and version 1 of the eBook and learning modules, Engineering Across Cultures, is due to be released in late October 2012. CONCLUSIONS The Engineering Across Cultures eBook and learning modules provide a useful and ready to employ resource to help educators tackle the complex issue of intercultural competency in engineering education. The book is by no means exhaustive, and nor are the modules, they instead provide an accessible, engineering specific guide to bringing cultural issues into the engineering classroom.

How fit is your organizational culture for business process management?

While business transformations often primarily focus on technological and methodological solutions, there is consensus that having the right organizational culture is critical for the successful change of business processes.

Effect of bone morphogenetic protein-4 (BMP-4) on the expression of Sox2, Oct-4 and c-Myc in human periodontal ligament cells during long-term culture

Recent studies demonstrated endogenous expression level of Sox2, Oct-4 and c-Myc is correlated with the pluripotency and successful induction of induced pluripotent stem cells (iPSCs). Periondontal ligament cells (PDLCs)have multi-lineage diferentiation capability and ability to maintain undifferentiated stage, which makes PDLCs a suitable cell source for tissue repair and regeneration. To elucidate the effect of in vitro culture condition on the stemness potential of PDLCs, we explored the cell growth, proliferation, cell cycle, and the expression of Sox2, Oct-4 and c-Myc in PDLCs from passage 1 to 7 with or without the addition of recombinant human BMP4(rhBMP4). Our results revealed that BMP-4 promoted cell growth and proliferation, arrested PDLCs in S phase of cell cycle and upregulated PI value. It was revealed that without the addition of rhBMP4, the expression of Sox2, Oct-4 and c-Myc in PDLCs only maintained nucleus location until passage 3, then lost nucleus location subsequently. The mRNA expression in PDLCs further confirmed that the level of Sox2 and Oct-4 peaked at passage 3, then decreased afterwards, whereas c-Myc maintained consistently upregulation along passages. after the treatment with rhBMP4, the expression of Sox2, Oct-4 and c-Myc in PDLCs maintained nucleus location even at passage 7 and the mRNA expression of Sox2 and Oct-4 significantly upregulated at passage 5 and 7. These results demonstrated that addition of rhBMP-4 in the culture media could improve the current culture condition for PDLCs to maintain in an undifferentiated stage.

Organizational culture and willingness to share knowledge : a competing values perspective in Australian context

A considerable amount of research has confirmed the relationship between organizational culture and knowledge sharing behaviours. However, less research has been conducted on the impact of project sub-cultures in relation to the sharing of knowledge between projects, particularly in project based organizations (PBOs). The unique structures and contexts characterized by PBOs indicate the need to investigate further the impact of cultures present within PBOs and their effect on knowledge sharing. We report on a rich case study of four large Australian-based PBOs whereby the cultural values of these large organizations were seen to impact significantly on whether project teams were more or less likely to improve inter-project knowledge sharing. Furthermore, this research demonstrates the utility of using Cameron and Quinn's (2005) Competing Values Framework to evaluate culture in the context of PBOs