Tissue engineering of stratified articular cartilage from chondrocyte subpopulations

Objective: To test if subpopulations of chondrocytes from different cartilage zones could be used to engineer cartilage constructs with features of normal stratification. Design: Chondrocytes from the superficial and middle zones of immature bovine cartilage were cultured in alginate, released, and seeded either separately or sequentially to form cartilage constructs. Constructs were cultured for 1 or 2 weeks and were assessed for growth, compressive properties, and deposition, and localization of matrix molecules and superficial zone protein (SZP). Results: The cartilaginous constructs formed from superficial zone chondrocytes exhibited less matrix growth and lower compressive properties than constructs from middle zone chondrocytes, with the stratified superficial-middle constructs exhibiting intermediate properties. Expression of SZP was highest at the construct surfaces, with the localization of SZP in superficial-middle constructs being concentrated at the superficial surface. Conclusions: Manipulation of subpopulations of chondrocytes can be useful in engineering cartilage tissue with a biomimetic approach, and in fabricating constructs that exhibit stratified features of normal articular cartilage. (C) 2003 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

Synthesis of proteoglycan 4 by chondrocyte subpopulations in cartilage explants, monolayer cultures, and resurfaced cartilage cultures

Objective: To quantify the levels of proteoglycan 4 (PRG4) expression by subpopulations of chondrocytes from superficial, middle, and deep layers of normal bovine calf cartilage in various culture systems. Methods: Bovine calf articular cartilage discs or isolated cells were used in I of 3 systems of chondrocyte culture: explant, monolayer, or transplant, for 1-9 days. PRG4 expression was quantified by enzyme-linked immunosorbent assay of spent medium and localized by immunohistochemistry at the articular surface and within chondrocytes in explants and cultured cells. Results: Superficial chondrocytes secreted much more PRG4 than did middle and deep chondrocytes in all cultures. The pattern of PRG4 secretion into superficial culture medium varied with the duration of culture, decreasing with time in explant culture (from similar to25 mug/cm(2)/day on days 0-1 to similar to3 mug/cm(2)/day on days 5-9), while increasing in monolayer culture (from similar to1 pg/cell/day on days 0-1 to similar to7 pg/cell/day on days 7-9) and tending to increase in transplant culture (reaching similar to2 mug/cm(2)/day by days 7-9). In all of the culture systems, inclusion of ascorbic acid stimulated PRG4 secretion, and the source of PRG4 was immunolocalized to superficial cells. Conclusion: The results described here indicate that the phenotype of PRG4 secretion by chondrocytes in culture is generally maintained, in that PRG4 is expressed to a much greater degree by chondrocytes from the superficial zone than by those from the middle and deep zones. The marked up-regulation of PRG4 synthesis by ascorbic acid may have implications for cartilage homeostasis and prevention of osteoarthritic disease. Transplanting specialized cells that secrete PRG4 to a surface may impart functional lubrication and be generally applicable to many tissues in the body.

Cell density alters matrix accumulation in two distinct fractions and the mechanical integrity of alginate-chondrocyte constructs

Chondrocyte density in articular cartilage is known to change with the development and growth of the tissue and may play an important role in the formation of a functional extracellular matrix (ECM). The objective of this study was to determine how initial chondrocyte density in an alginate hydrogel affects the matrix composition, its distribution between the cell-associated (CM) and further removed matrix (FRM) fractions, and the tensile mechanical properties of the developing engineered cartilage. Alginate constructs containing primary bovine chondrocytes at densities of 0, 4, 16, and 64 million cells/ml were fabricated and cultured for 1 or 2 weeks, at which time structural, biochemical, and mechanical properties were analyzed. Both matrix content and distribution varied with the initial cell density. Increasing cell density resulted in an increasing content of collagen and sulfated-glycosaminoglycan (GAG) and an increasing proportion of these molecules localized in the CM. While the equilibrium tensile modulus of cell-free alginate did not change with time in culture, the constructs with highest cell density were 116% stiffer than cell-free controls after 2 weeks of culture. The equilibrium tensile modulus was positively correlated with total collagen (r2 = 0.47, p < 0.001) and GAG content (r2 = 0.68, p < 0.001), and these relationships were enhanced when analyzing only those matrix molecules in the CM fraction (r2 = 0.60 and 0.72 for collagen and GAG, respectively, each p < 0.001). Overall, the results of this study indicate that initial cell density has a considerable effect on the developing composition, structure, and function of alginate–chondrocyte constructs.

Short-term retention of labeled chondrocyte subpopulations in stratified tissue-engineered cartilaginous constructs implanted in vivo in mini-pigs

It is likely that effective application of cell-laden implants for cartilage defects depends on retention of implanted cells and interaction between implanted and host cells. The objectives of this study were to characterize stratified cartilaginous constructs seeded sequentially with superficial (S) and middle (M) chondrocyte subpopulations labelled with fluorescent cell tracking dye PKH26 (*) and determine the degree to which these stratified cartilaginous constructs maintain their architecture in vivo after implantation in mini-pigs for 1 week. Alginate-recovered cells were seeded sequentially to form stratified S*/M (only S cells labelled) and S*/M* (both S and M cells labelled) constructs. Full-thickness defects (4 mm diameter) were created in the patellofemoral groove of adult Yucatan mini-pigs and filled with portions of constructs or left empty. Constructs were characterized biochemically, histologically, and biomechanically, and stratification visualized and quantified, before and after implant. After 1 week, animals were sacrificed and implants retrieved. After 1 week in vivo, glycosaminoglycan and collagen content of constructs remained similar to that at implant, whereas DNA content increased. Histological analyses revealed features of an early repair response, with defects filled with tissues containing little matrix and abundant cells. Some implanted (PKH26-labeled) cells persisted in the defects, although constructs did not maintain a stratified organization. Of the labelled cells, 126 +/- 38% and 32 +/- 8% in S*/M and S*/M* constructs, respectively, were recovered. Distribution of labelled cells indicated interactions between implanted and host cells. Longer-term in vivo studies will be useful in determining whether implanted cells are sufficient to have a positive effect in repair.

Depth-dependent biomechanical and biochemical properties of fetal, newborn, and tissue-engineered articular cartilage

Adult articular cartilage has depth-dependent mechanical and biochemical properties which contribute to zone-specific functions. The compressive moduli of immature cartilage and tissue-engineered cartilage are known to be lower than those of adult cartilage. The objective of this study was to determine if such tissues exhibit depth-dependent compressive properties, and how these depth-varying properties were correlated with cell and matrix composition of the tissue. The compressive moduli of fetal and newborn bovine articular cartilage increased with depth (p < 0.05) by a factor of 4-5 from the top 0.1 mm (28 +/- 13 kPa, 141 +/- 10 kPa, respectively) to 1 mm deep into the tissue. Likewise, the glycosaminoglycan and collagen content increased with depth (both p < 0.001), and correlated with the modulus (both p < 0.01). In contrast, tissue-engineered cartilage formed by either layering or mixing cells from the superficial and middle zone of articular cartilage exhibited similarly soft regions at both construct surfaces, as exemplified by large equilibrium strains. The properties of immature cartilage may provide a template for developing tissue-engineered cartilage which aims to repair cartilage defects by recapitulating the natural development and growth processes. These results suggest that while depth-dependent properties may be important to engineer into cartilage constructs, issues other than cell heterogeneity must be addressed to generate such tissues. (c) 2005 Elsevier Ltd. All rights reserved.

Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in Vitro redifferentiation

Background: If chondrocytes from the superficial, middle, and deep zones of articular cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. ----- ----- Hypothesis: Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture).----- ----- Study Design: Controlled laboratory study.----- ----- Methods: Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes).----- ----- Results: Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone.----- ----- Conclusion: Chondrocytes isolated from the 3 zones of equine cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion.

Queering Paradigms II : Interrogating Agendas

This book offers a fundamental challenge to a variety of theoretical, social, and political paradigms, ranging from law and justice studies to popular culture, linguistics to political activism. Developing the intellectual project initiated in Queering Paradigms, this volume extends queer theorizing in challenging new directions and uses queer insights to explore, trouble, and interrogate the social, political, and intellectual agendas that pervade (and are often taken for granted within) public discourses and academic disciplines. The contributing authors include queer theorists, socio-linguists, sociologists, political activists, educators, social workers and criminologists. Together, they contribute not only to the ongoing process of theorizing queerly, but also to the critique and reformulation of their respective disciplines.

Portrayal of librarians in Australian creative writing

An exploration is made of the ways in which librarians have been depicted in Australian creative writing. Reference is made to characters in novels, short stories, drama and poetry. With respect to novels, there is some consideration of characterisation and its relationship to plot.

Information and communication technology for E-regions

Information and communication technologies (ICTs) are essential components of the knowledge economy, and have an immense complementary role in innovation, education, knowledge creation, and relations with government, civil society, and business within city regions. The ability to create, distribute, and exploit knowledge has become a major source of competitive advantage, wealth creation, and improvements in the new regional policies. Growing impact of ICTs on the economy and society, rapid application of recent scientific advances in new products and processes, shifting to more knowledge-intensive industry and services, and rising skill requirements have become crucial concepts for urban and regional competitiveness. Therefore, harnessing ICTs for knowledge-based urban development (KBUD) has a significant impact on urban and regional growth (Yigitcanlar, 2005). In this sense, e-region is a novel concept utilizing ICTs for regional development. Since the Helsinki European Council announced Turkey as a candidate for European Union (EU) membership in 1999, the candidacy has accelerated the speed of regional policy enhancements and adoption of the European regional policy standards. These enhancements and adoption include the generation of a new regional spatial division, NUTS-II statistical regions; a new legislation on the establishment of regional development agencies (RDAs); and new orientations in the field of high education, science, and technology within the framework of the EU’s Lisbon Strategy and the Bologna Process. The European standards posed an ambitious new agenda in the development and application of contemporary regional policy in Turkey (Bilen, 2005). In this sense, novel regional policies in Turkey necessarily endeavor to include information society objectives through efficient use of new technologies such as ICTs. Such a development seeks to be based on tangible assets of the region (Friedmann, 2006) as well as the best practices deriving from grounding initiatives on urban and local levels. These assets provide the foundation of an e-region that harnesses regional development in an information society context. With successful implementations, the Marmara region’s local governments in Turkey are setting the benchmark for the country in the implementation of spatial information systems and e-governance, and moving toward an e-region. Therefore, this article aims to shed light on organizational and regional realities of recent practices of ICT applications and their supply instruments based on evidence from selected local government organizations in the Marmara region. This article also exemplifies challenges and opportunities of the region in moving toward an e-region and provides a concise review of different ICT applications and strategies in a broader urban and regional context. The article is organized in three parts. The following section scrutinizes the e-region framework and the role of ICTs in regional development. Then, Marmara’s opportunities and challenges in moving toward an e-region are discussed in the context of ICT applications and their supply instruments based on public-sector projects, policies, and initiatives. Subsequently, the last section discusses conclusions and prospective research.

Optimization of container process at seaport terminals

Seaport container terminals are an important part of the logistics systems in international trades. This paper investigates the relationship between quay cranes, yard machines and container storage locations in a multi-berth and multi-ship environment. The aims are to develop a model for improving the operation efficiency of the seaports and to develop an analytical tool for yard operation planning. Due to the fact that the container transfer times are sequence-dependent and with the large number of variables involve, the proposed model cannot be solved in a reasonable time interval for realistically sized problems. For this reason, List Scheduling and Tabu Search algorithms have been developed to solve this formidable and NP-hard scheduling problem. Numerical implementations have been analysed and promising results have been achieved.

Analysis of build-up of heavy metals and volatile organics on urban roads in Gold Coast, Australia

Urban water quality can be significantly impaired by the build-up of pollutants such as heavy metals and volatile organics on urban road surfaces due to vehicular traffic. Any control strategy for the mitigation of traffic related build-up of heavy metals and volatile organic pollutants should be based on the knowledge of their build-up processes. In the study discussed in this paper, the outcomes of a detailed experiment investigation into build-up processes of heavy metals and volatile organics are presented. It was found that traffic parameters such as average daily traffic, volume over capacity ratio and surface texture depth had similar strong correlations with the build-up of heavy metals and volatile organics. Multicriteria decision analyses revealed that the 1 - 74 um particulate fraction of total suspended solids (TSS) could be regarded as a surrogate indicator for particulate heavy metals in build-up and this same fraction of total organic carbon could be regarded as a surrogate indicator for particulate volatile organics build-up. In terms of pollutants affinity, TSS was found to be the predominant parameter for particulate heavy metals build-up and total dissolved solids was found to be the predominant parameter for he potential dissolved particulate fraction in heavy metals build-up. It was also found that land use did not play a significant role in the build-up of traffic generated heavy metals and volatile organics.

International regional planning educational experience : learnings from Malaysia, Korea and Turkey study trips

In an era of rapidly changing economic, social and environmental conditions, urban and regional planning education must be resilient, innovative and able to deal with the complex political and socio-economic fabric of post-modern cities. As a consequence, urban and regional planning education plays a fundamental role in educating and forming planning practitioners that will be able to tackle such complexity. However, not many tertiary education institutions provide a trans-cultural engagement opportunity for students, where the need to internationalise planning education has been widely recognised worldwide. The aim of this paper is to communicate the findings of three overseas study trips (Kuala Lumpur-Malaysia, Daejeon-Korea, Istanbul and Gallipoli-Turkey) that students of Queensland University of Technology are taken to where these study trips trailed the provision of an innovative tertiary education experience of teaching regional planning in an international context. The findings of the pedagogic analyses of the study reveal that the exposure of students to different planning processes and practices give them a new outlook on what they knew from their own country and provide them with useful insights on international planning issues and cultural differences and barriers.

Enhancing professional ethics : service-learning in teacher education

The ethical conduct of professionals has been the focus of increasing scrutiny over the past several decades as members of the public, the media, professional bodies, and legislative authorities have struggled to define ethical behaviour in times of governmental change, increasing internationalisation, globalised communications, threats of terrorism, and the challenges of developments in science and medicine (e.g., Demmke & Bossaert, 2004). National governments and transnational bodies have responded to these concerns about ethics and corruption through measures such as the United Nations Convention Against Corruption (United Nations Office on Drugs and Crime, 2004), Transparency International’s annual corruption index (2010) and Queensland’s Public Sector Ethics Act 1994 (Queensland Parliament 1994). Similarly, academic interest in ethics and its application across a range of domains(e.g., business, health care, social welfare, criminal justice, law, journalism, defence, environment, and media) has also increased. To illustrate, in 1993, a non-partisan, non-profit national umbrella organisation, the Australian Association for Professional and Applied Ethics, was formed following a conference concerned with the teaching of ethics (http://www.arts.unsw.edu.au./aapae/about_aapae/about_aapae.htm), while a recent review of the Excellence in Research for Australian rankings of national and international academic journals revealed that 16 journals related to ethics had received the top ratings of A* or A (Australian Research Council, 2009). In this chapter we examine professional ethics and argue, with specific reference to the context of pre-service teacher education, that Service-learning is one way of enhancing emerging professionals’ understanding of ethics.

Australian Library and Information Studies (LIS) researchers' ranking of LIS journals

The paper describes the processes and the outcomes of the ranking of LIS journal titles by Australia’s LIS researchers during 2007-8, firstly through the Australian federal government’s Research Quality Framework (RQF) process and then its replacement, the Excellence in Research for Australia (ERA) initiative. The requirement to rank the journals titles used came from discussions held at the RQF panel meeting held in February 2007 in Canberra, Australia. While it was recognised that the Web of Science (formerly ISI) journal impact approach of journal acceptance for measures of research quality and impact might not work for LIS, it was apparent that this model would be the default if no other ranking of journal titles became apparent. Although an increasing number of LIS and related discipline journals were appearing in the Web of Science listed rankings, the number was few and it was thus decided by the Australian LIS research community to undertake the ranking exercise.

The Secret Siphon

Although the siphon has been in use since ancient times, the exact mechanism of operation is still under discussion. For example, most dictionaries assert that atmospheric pressure is essential to the operation of a siphon rather than gravity. Although there is general agreement that gravity is the motivating force in a siphon, there is disagreement on how liquid enters a siphon – is it atmospheric push or tensile pull? This paper describes a classroom experiment that can serve as the basis for discussing how a siphon works. The experiment involves the construction of a siphon in which the water level in the upper reservoir is held constant during the operation of the siphon. Since the atmosphere is not doing any work on the water in the upper reservoir only gravity is at work. The special situation of a bubble-in-a-siphon is also discussed in which both atmospheric pressure and gravity are at work.