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.

Uncovering the evidence of non-expert nephrology nursing practice

Expertise in nursing has been widely studied although there have been no previous studies into what constitutes expertise in nephrology (renal) nursing. This paper, which is abstracted from a larger study into the acquisition and exercise of nephrology nursing expertise, provides evidence of the characteristics and practices of non-expert nephrology nurses. Using the grounded theory method, the study took place in one renal unit in New South Wales, Australia, and involved six non-expert and 11 expert nurses. Sampling was purposive then theoretical. Simultaneous data collection and analysis using participant observation, review of nursing documentation and semistructured interviews was undertaken. The study revealed a three-stage skills-acquisitive process that was identified as non-expert, experienced non-expert and expert stages. Non-expert nurses showed superficial nephrology nursing knowledge and limited experience; they were acquiring basic nephrology nursing skills and possessed a narrow focus of practice.

Flow regime transition criteria for two-phase flow at reduced gravity conditions

Flow regime transition criteria are of practical importance for two-phase flow analyses at reduced gravity conditions. Here, flow regime transition criteria which take the friction pressure loss effect into account were studied in detail. Criteria at reduced gravity conditions were developed by extending an existing model with various experimental datasets taken at microgravity conditions showed satisfactory agreement. Sample computations of the model were performed at various gravity conditions, such as 0.196, 1.62, 3.71, and 9.81 m/s2 corresponding to micro-gravity and lunar, Martian and Earth surface gravity, respectively. It was found that the effect of gravity on bubbly-slug and slug-annular (churn) transitions in a two-phase flow system was more pronounced at low liquid flow conditions, whereas the gravity effect could be ignored at high mixture volumetric flux conditions. While for the annular flow transitions due to flow reversal and onset of dropset entrainment, higher superficial gas velocity was obtained at higher gravity level.

Research process management

Though the value of a process-centred view for the understanding and (re-)design of corporations has been widely accepted, our understanding of the research process in Information Systems (IS) remains superficial. A process-centred view on IS research considers the conduct of a research project as a sequence of activities involving resources, data and research artifacts. As such, it helps to reflect on more effective ways to conduct IS research, to consolidate and compare diverse practices and to complement the focus on research methodologies with research project practices. This paper takes a first step towards the discipline of ‘Research Process Management’ by exploring the features of research processes and by presenting a preliminary approach for research process design that can facilitate modelling IS research. The case study method and the design science research method are used as examples to demonstrate the potential of such reference research process models.

Something rotten in the state of Minnesota: or the morality of backwoodsmen: 'A Simple Plan'

'A Simple Plan' is a deceptively complex and multilayered film, combining elements of Celtic mythology with the morality play and the windfall fantasy gone disastrously wrong. Despite its blending of realism and heavyhanded symbolism, and its abundant trans-textual gestures, 'A Simple Plan' is in many ways defiantly not a 90s movie: its leading characters are fashionably flawed, but they are neither sensitive, nor honourable, nor heroic; there are no startling special effects or intricate timeshifts; and it desperately gives the impression of depth, of being emphatically more than mere superficial excess. At a stretch it almost appears to be a throwback to the 1930s Production Code emphasis on the role of cinema in moral instruction; while good hardly triumphs over evil, venality is painfully and emphatically punished. But in other ways it is a quintessential late 90s film: an American/British/Japanese/German/French co-production, 'A Simple Plan' acts most palpably as a commentary on the moral, economic and social condition of the United States at the end of the American century.

Effects of oxygen on zonal marker expression in human articular chondrocytes

Articular cartilage is organized in depth zones with phenotypically distinct subpopulations of chondrocytes that are exposed to different oxygen tensions. Despite growing evidence of the critical role for oxygen in chondrogenesis, little is known about its effect on chondrocytes from different zones. This study evaluates zonal marker expression of human articular chondrocytes from different zones under various oxygen tensions. Chondrocytes isolated from full-thickness, superficial, and middle/deep cartilage from knee replacement surgeries were expanded and redifferentiated under hypoxic (5% O 2) or normoxic (20% O 2) conditions. Differentiation under hypoxia increased expression of hypoxia-inducible factors 1alpha and 2alpha and accumulation of extracellular matrix, particularly in middle/deep chondrocytes, and favored re-expression of proteoglycan 4 by superficial chondrocytes compared with middle/deep cells. Zone-dependent expression of clusterin varied with culture duration. These results demonstrate that zonal chondrocytes retain important phenotypic differences during in vitro cultivation, and that these characteristics can be improved by altering the oxygen environment. However, transcript levels for pleiotrophin, cartilage intermediate layer protein, and collagen type X were similar between zones, challenging their reliability as zonal markers for tissue-engineered cartilage from osteoarthritis patients. Key factors including oxygen tension and cell source should be considered to prescribe zone-specific properties to tissue-engineered cartilage. © 2012, Mary Ann Liebert, Inc.

Dynamic compression improves biosynthesis of human zonal chondrocytes from osteoarthritis patients

Objective: We hypothesize that chondrocytes from distinct zones of articular cartilage respond differently to compressive loading, and that zonal chondrocytes from osteoarthritis (OA) patients can benefit from optimized compressive stimulation. Therefore, we aimed to determine the transcriptional response of superficial (S) and middle/deep (MD) zone chondrocytes to varying dynamic compressive strain and loading duration. To confirm effects of compressive stimulation on overall matrix production, we subjected zonal chondrocytes to compression for 2 weeks. Design: Human S and MD chondrocytes from osteoarthritic joints were encapsulated in 2% alginate, pre-cultured, and subjected to compression with varying dynamic strain (5, 15, 50% at 1 Hz) and loading duration (1, 3, 12 h). Temporal changes in cartilage-specific, zonal, and dedifferentiation genes following compression were evaluated using quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR). The benefits of long-term compression (50% strain, 3 h/day, for 2 weeks) were assessed by measuring construct glycosaminoglycan (GAG) content and compressive moduli, as well as immunostaining. Results: Compressive stimulation significantly induced aggrecan (ACAN), COL2A1, COL1A1, proteoglycan 4 (PRG4), and COL10A1 gene expression after 2 h of unloading, in a zone-dependent manner (P < 0.05). ACAN and PRG4 mRNA levels depended on strain and load duration, with 50% and 3 h loading resulting in highest levels (P < 0.05). Long-term compression increased collagen type II and ACAN immunostaining and total GAG (P < 0.05), but only S constructs showed more PRG4 stain, retained more GAG (P < 0.01), and developed higher compressive moduli than non-loaded controls. Conclusions: The biosynthetic activity of zonal chondrocytes from osteoarthritis joints can be enhanced with selected compression regimes, indicating the potential for cartilage tissue engineering applications. © 2012 Osteoarthritis Research Society International.

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.

Multimodal approaches to reflective teaching and assessment in higher education

Reflection is not a new concept in the teaching of higher education and is often an important component of many disciplinary courses. Despite this, past research shows that whilst there are examples of rich reflective strategies used in some areas of higher education, most approaches to and conceptualisations of reflective learning and assessment have been perfunctory and inconsistent. In many disciplinary areas reflection is often assessed as a written activity ‘tagged onto’ assessment practices. In creative disciplines however, reflective practice is an integral and cumulative form of learning and is often expressed in ways other than in the written form. This paper will present three case studies of reflective practice in the area of Creative Industries in higher education – Dance, Fashion and Music. It will discuss the ways in which higher education teachers and students use multi-modal approaches to expressing knowledge and reflective practice in context. The paper will argue that unless students are encouraged to participate in deep reflective disciplinary discourse via multi-modes then reflection will remain superficial in the higher education context.

Complications related to therapeutic anticoagulation in total hip arthroplasty

Bleeding related wound complications cause significant morbidity in lower limb arthroplasty surgery. Patients who require therapeutic anticoagulation in the peri operative period are potentially at higher risk of these complications. This is a retrospective case control study reviewing all primary total hip replacements performed in a single center over a five year period and comparing outcomes of the patients on warfarin with a double-matched control group of patients not on warfarin. The warfarin group had significantly higher risk of deep joint infection (9% vs 2.2%), haematoma/ wound ooze (28% vs 4%) and superficial infection (13.5% vs 2.2%). Managing the total hip arthroplasty patient with therapeutic anticoagulation is a balance between the risk of thromboembolic disease and bleeding related complications. Improved understanding of this risk will better allow the patient to make an informed decision regarding their elective arthroplasty surgery.

Achieving at university and beyond : does it help to be good looking?

Purpose – The purpose of this study is to examine the associations between attitudes to academic achievement and post university success using perceptions of attractiveness, gender, ethnic identity, personality, and social acceptance as antecedents. Design/methodology/approach – An online questionnaire was completed by male (N=116) and female (N=126) university students from various cultural backgrounds. To evaluate the proposed relationships, multiple regression analysis was used. Findings – The findings suggest that attractiveness is related to attitudes to academic achievement and success through its association with social appeal and acceptance. Ethnic identity is also related to both academic achievement and post university success. Personality is not positively related to academic achievement. Finally, social acceptance is positively related to academic achievement for males and to success for females. Research limitations/implications – Whilst the survey targeted students from various cultural backgrounds studying in Australia, it did not look at university students from other countries. A cross-cultural perspective could reveal further differences in attitudes. Originality/value – This study links attractiveness and academic achievement theories. The findings have implications for tertiary institutions and suggest academics and policy-makers to vigorously promote core personality and values such as intelligence, communication skills, and sincerity, rather than allow superficial values such as attractiveness to be placed at the centre stage of students' endeavour.

Re-framing fashion : from original and copy to adaptation

Partington notes that clothing produced by individual consumers through adaptation of patterns is contextualised as a watered down version of original couture. In its most reductive form, this notion characterises fashion as commercial and exploitative. Descriptors such as appropriation, imitation, copy and so forth have restricted the opportunity to understand fashion as a major global cultural form and institution. Therefore exploring and understanding the concept of adaptation will shift the attention from a superficial assessment of original versus imitation or copy to adaptation as a practice that provides a better framework for the understanding of designers’ and couturiers’ innovative practices and creativity, describing also the active engagement of consumers with fashion at the micro level. Adaptation can also provide a way to understand different historical shifts in the fashion system, from individual creative agency with home dressmaking and re-making to the explosion of the mass market and the consequent abandonment of such practices. Home dressmaking has been replaced by fashion remix of mass produced garments, a practice that thrives in our environment of globalised fast fashion. Thus this chapter suggests the need for a contextual requalification of concepts such as original, copy, imitation and copyright, and argues that these categories have been played against each other, but they are in fact interdependent. Today, big labels and conglomerates try to control knowledge and innovation through copyright, but, fashion escapes copyright because, in fashion, creativity is contextual. The institutionalisation of couture from 1868 served as a way to control knowledge about production processes in fashion; on the other hand, adaptation practices, often subversive, have been fundamental to the democratisation of fashion.

Varianza analitica y aproximada del costo total de un proyecto

La varianza estadística del costo total de un proyecto usualmente se estima por medio de la simulación de Monte Carlo, bajo el supuesto de que los acercamientos analíticos son demasiado complicados. Este artículo analiza este supuesto y muestra que, contrario a lo esperado, la solución analítica es relativamente directa. También se muestra que el coeficiente de variación no se ve afectado por el tamaño (área superficial) del proyecto cuando se usan los costos de los componentes estandarizados. Se provee un caso de estudio en el cual se analizan los costos reales de los componentes para obtener la varianza del costo total requerida. Los resultados confirman trabajos previos al mostrar que la aproximación del segundo momento (varianza) bajo el supuesto de independencia subestima considerablemente el valor exacto. El análisis continua examinando los efectos del juicio profesional y con los datos simulados utilizados, la aproximación resulta razonablemente exacta - el juicio profesional absorbe la mayor parte de las intercorrelaciones involucradas. También se da un ejemplo en el cual las cantidades de los componentes unitarios son valoradas por sus costos unitarios promedios y muestra, una vez más, que la aproximación es cercana al valor real. Finalmente, el trabajo se extiende mostrando cómo obtener, para cada proyecto, las varianzas exactas del costo total.