Vitrification as a prospect for cryopreservation of tissue-engineered constructs

Cryopreservation plays a significant function in tissue banking and will presume yet larger value when more and more tissue-engineered products will routinely enter the clinical arena. The most common concept underlying tissue engineering is to combine a scaffold (cellular solids) or matrix (hydrogels) with living cells to form a tissue-engineered construct (TEC) to promote the repair and regeneration of tissues. The scaffold and matrix are expected to support cell colonization, migration, growth and differentiation, and to guide the development of the required tissue. The promises of tissue engineering, however, depend on the ability to physically distribute the products to patients in need. For this reason, the ability to cryogenically preserve not only cells, but also TECs, and one day even whole laboratory-produced organs, may be indispensable. Cryopreservation can be achieved by conventional freezing and vitrification (ice-free cryopreservation). In this publication we try to define the needs versus the desires of vitrifying TECs, with particular emphasis on the cryoprotectant properties, suitable materials and morphology. It is concluded that the formation of ice, through both direct and indirect effects, is probably fundamental to these difficulties, and this is why vitrification seems to be the most promising modality of cryopreservation

The Australian Grey Nomads : are they who we think they are? Enhancing formative research through the quantitative assessment of psychological constructs

Issue addressed: Measures of 'social identity' and 'psychological sense of community' were included within a broader formative research inquiry to gain insight into the identity characteristics and level of connectedness among older recreational road travellers (commonly known as Grey Nomads). The research sought to gain insights on how best to reach or speak to this growing driver cohort. ----- ----- Method: Participants included 631 older recreational road travellers ranging in age from 50 years to over 80 years. Data were obtained through three scales which were incorporated into a larger formative research survey; an identity hierarchy, the Three Factor Model of Social Identity and the Sense of Community Index. ----- ----- Results: Older recreational road travellers see themselves principally as couples, with social group identity being secondary. Although many identified to some degree with the Grey Nomad identity, when asked to self categorise as either members of the Broad Network of Recreational Vehicle Travellers or as Grey Nomads, the majority categorised themselves as the former. Those identifying as Grey Nomads, however, reported significantly higher levels of 'social identification' and 'sense of community'. ----- ----- Conclusion: The Grey Nomad identity may not be the best identity at which to target road safety messages for this cohort. Targeting travelling 'couples' may be more efficacious. Using the 'Grey Nomad' identity is likely to reap at least some success, however, given that many identified to some degree with this group identity. Those identifying as Grey Nomads may be more open to community participation or behaviour change given their significantly higher levels of 'social identity' and 'sense of community'.

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.

Performance analysis of business process models with advanced constructs

The importance of actively managing and analysing business processes is acknowledged more than ever in organisations nowadays. Business processes form an essential part of an organisation and their application areas are manifold. Most organisations keep records of various activities that have been carried out for auditing purposes, but they are rarely used for analysis purposes. This paper describes the design and implementation of a process analysis tool that replays, analyses and visualises a variety of performance metrics using a process definition and its corresponding execution logs. The replayer uses a YAWL process model example to demonstrate its capacity to support advanced language constructs.

The dynamics of parenting and early feeding - constructs and controversies : a viewpoint

There is increasing evidence that parenting and feeding interact to influence children’s eating behaviour and weight status. Interpretation of existing research is complicated by the lack of consensus in the conceptualisation and measurement of both ‘parenting’ and ‘feeding’, particularly the distinction between ‘styles’, ‘dimensions’ and ‘practices’. In addition, the lack of validated tools to concurrently assess feeding practices in infancy limits the capacity to examine the relationships between parenting and feeding in infancy and their short- and long-term influence on weight status. In this paper we provide an overview of the constructs examined in this emerging area of research, highlight the conceptual, definitional and measurement challenges and propose a unifying model to aid design and the interpretation of intervention studies. Progress on these methodological issues will contribute to the robust evidence required to justify investment in interventions that focus on parenting and feeding in the context of child obesity prevention.

Robust performance analysis on YAWL process models with advanced constructs

The importance of actively managing and analyzing business processes is acknowledged more than ever in organizations nowadays. Business processes form an essential part of an organization and their ap-plication areas are manifold. Most organizations keep records of various activities that have been carried out for auditing purposes, but they are rarely used for analysis purposes. This paper describes the design and implementation of a process analysis tool that replays, analyzes and visualizes a variety of performance metrics using a process definition and its execution logs. Performing performance analysis on existing and planned process models offers a great way for organizations to detect bottlenecks within their processes and allow them to make more effective process improvement decisions. Our technique is applied to processes modeled in the YAWL language. Execution logs of process instances are compared against the corresponding YAWL process model and replayed in a robust manner, taking into account any noise in the logs. Finally, performance characteristics, obtained from replaying the log in the model, are projected onto the model.

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.

Engineering of vascularized adipose constructs

Adipose tissue engineering offers a promising alternative to the current surgical techniques for the treatment of soft tissue defects. It is a challenge to find the appropriate scaffold that not only represents a suitable environment for cells but also allows fabrication of customized tissue constructs, particularly in breast surgery. We investigated two different scaffolds for their potential use in adipose tissue regeneration. Sponge-like polyurethane scaffolds were prepared by mold casting with methylal as foaming agent, whereas polycaprolactone scaffolds with highly regular stacked-fiber architecture were fabricated with fused deposition modeling. Both scaffold types were seeded with human adipose tissuederived precursor cells, cultured and implanted in nude mice using a femoral arteriovenous flow-through vessel loop for angiogenesis. In vitro, cells attached to both scaffolds and differentiated into adipocytes. In vivo, angiogenesis and adipose tissue formation were observed throughout both constructs after 2 and 4 weeks, with angiogenesis being comparable in seeded and unseeded constructs. Fibrous tissue formation and adipogenesis were more pronounced on polyurethane foam scaffolds than on polycaprolactone prototyped scaffolds. In conclusion, both scaffold designs can be effectively used for adipose tissue engineering.

Measurement constructs to explore innovation diffusion in construction

Although the drivers of innovation have been studied extensively in construction, greater attention is required on how innovation diffusion can be effectively assessed within this complex and interdependent project-based industry. The authors draw on a highly cited innovation diffusion model by Rogers (2006) and develop a tailored conceptual framework to guide future empirical work aimed at assessing innovation diffusion in construction. The conceptual framework developed and discussed in this paper supports a five-stage process model of innovation diffusion namely: 1) knowledge and idea generation, 2) persuasion and evaluation; 3) decision to adopt, 4) integration and implementation, and 5) confirmation. As its theoretical contribution, this paper proposes three critical measurements constructs which can be used to assess the effectiveness of the diffusion process. These measurement constructs comprise: 1) nature and introduction of an innovative idea, 2) organizational capacity to acquire, assimilate, transform and exploit an innovation, and 3) rates of innovation facilitation and adoption. The constructs are interpreted in the project-based context of the construction industry, extending the contribution of general management theorists. Research planned by the authors will test the validity and reliability of the constructs developed in this paper.