Expansion and chondrogenic potential of human articular chondrocytes on macroporous microcarriers

Regenerative medicine techniques are currently being investigated to replace damaged cartilage. Critical to the success of these techniques is the ability to expand the initial population of cells while minimising de-differentiation to allow for hyaline cartilage to form. Three-dimensional culture systems have been shown to enhance the differentiation of chondrocytes in comparison to two-dimensional culture systems. Additionally, bioreactor expansion on microcarriers can provide mechanical stimulation and reduce the amount of cellular manipulation during expansion. The aim of this study was to characterise the expansion of human chondrocytes on microcarriers and to determine their potential to form cartilaginous tissue in vitro. High-grade human articular cartilage was obtained from leg amputations with ethics approval. Chondrocytes were isolated by collagenase digestion and expanded in either monolayers (104 cells/cm2) or on CultiSpher-G microcarriers (104 cells/mg) for three weeks. Following expansion, monolayer cells were passaged and cells on microcarriers were either left intact or the cells were released with trypsin/EDTA. Pellets from these three groups were formed and cultured for three weeks to establish the chondrogenic differentiation potential of monolayer-expanded and microcarrier-expanded chondrocytes. Cell viability, proliferation, glycosaminoglycan (GAG) accumulation, and collagen synthesis were assessed. Histology and immunohistochemistry were also performed. Human chondrocytes remained viable and expanded on microcarriers 10.2±2.6 fold in three weeks. GAG content significantly increased with time, with the majority of GAG found in the medium. Collagen production per nanogram DNA increased marginally during expansion. Histology revealed that chondrocytes were randomly distributed on microcarrier surfaces yet most pores remained cell free. Critically, human chondrocytes expanded on microcarriers maintained their ability to redifferentiate in pellet culture, as demonstrated by Safranin-O and collagen II staining. These data confirm the feasibility of microcarriers for passage-free cultivation of human articular chondrocytes. However, cell expansion needs to be improved, perhaps through growth factor supplementation, for clinical utility. Recent data indicate that cell-laden microcarriers can be used to seed fresh microcarriers, thereby increasing the expansion factor while minimising enzymatic passage.

Caveat Anicula! Beware of the Quiet Little Old Ladies : Demographics, Pharmacotherapy, Survival and Readmissions in a 10 Year Cohort of Heart Failure Patients with Preserved Systolic Function

Objective--To determine whether heart failure with preserved systolic function (HFPSF) has different natural history from left ventricular systolic dysfunction (LVSD). Design and setting--A retrospective analysis of 10 years of data (for patients admitted between 1 July 1994 and 30 June 2004, and with a study census date of 30 June 2005) routinely collected as part of clinical practice in a large tertiary referral hospital.Main outcome measures-- Sociodemographic characteristics, diagnostic features, comorbid conditions, pharmacotherapies, readmission rates and survival.Results--Of the 2961 patients admitted with chronic heart failure, 753 had echocardiograms available for this analysis. Of these, 189 (25%) had normal left ventricular size and systolic function. In comparison to patients with LVSD, those with HFPSF were more often female (62.4% v 38.5%; P = 0.001), had less social support, and were more likely to live in nursing homes (17.9% v 7.6%; P < 0.001), and had a greater prevalence of renal impairment (86.7% v 6.2%; P = 0.004), anaemia (34.3% v 6.3%; P = 0.013) and atrial fibrillation (51.3% v 47.1%; P = 0.008), but significantly less ischaemic heart disease (53.4% v 81.2%; P = 0.001). Patients with HFPSF were less likely to be prescribed an angiotensin-converting enzyme inhibitor (61.9% v 72.5%; P = 0.008); carvedilol was used more frequently in LVSD (1.5% v 8.8%; P < 0.001). Readmission rates were higher in the HFPSF group (median, 2 v 1.5 admissions; P = 0.032), particularly for malignancy (4.2% v 1.8%; P < 0.001) and anaemia (3.9% v 2.3%; P < 0.001). Both groups had the same poor survival rate (P = 0.912). Conclusions--Patients with HFPSF were predominantly older women with less social support and higher readmission rates for associated comorbid illnesses. We therefore propose that reduced survival in HFPSF may relate more to comorbid conditions than suboptimal cardiac management.

Using Association Rules to Solve the Cold-Start Problem in Recommender Systems

Recommender systems are widely used online to help users find other products, items etc that they may be interested in based on what is known about that user in their profile. Often however user profiles may be short on information and thus it is difficult for a recommender system to make quality recommendations. This problem is known as the cold-start problem. Here we investigate using association rules as a source of information to expand a user profile and thus avoid this problem. Our experiments show that it is possible to use association rules to noticeably improve the performance of a recommender system under the cold-start situation. Furthermore, we also show that the improvement in performance obtained can be achieved while using non-redundant rule sets. This shows that non-redundant rules do not cause a loss of information and are just as informative as a set of association rules that contain redundancy.

Young drivers’ optimism bias for accident risk and driving skill : accountability and insight experience manipulations

This study aimed to determine whether two brief, low cost interventions would reduce young drivers’ optimism bias for their driving skills and accident risk perceptions. This tendency for such drivers to perceive themselves as more skilful and less prone to driving accidents than their peers may lead to less engagement in precautionary driving behaviours and a greater engagement in more dangerous driving behaviour. 243 young drivers (aged 17 - 25 years) were randomly allocated to one of three groups: accountability, insight or control. All participants provided both overall and specific situation ratings of their driving skills and accident risk relative to a typical young driver. Prior to completing the questionnaire, those in the accountability condition were first advised that their driving skills and accident risk would be later assessed via a driving simulator. Those in the insight condition first underwent a difficult computer-based hazard perception task designed to provide participants with insight into their potential limitations when responding to hazards in difficult and unpredictable driving situations. Participants in the control condition completed only the questionnaire. Results showed that the accountability manipulation was effective in reducing optimism bias in terms of participants’ comparative ratings of their accident risk in specific situations, though only for less experienced drivers. In contrast, among more experienced males, participants in the insight condition showed greater optimism bias for overall accident risk than their counterparts in the accountability or control groups. There were no effects of the manipulations on drivers’ skills ratings. The differential effects of the two types of manipulations on optimism bias relating to one’s accident risk in different subgroups of the young driver sample highlight the importance of targeting interventions for different levels of experience. Accountability interventions may be beneficial for less experienced young drivers but the results suggest exercising caution with the use of insight type interventions, particularly hazard perception style tasks, for more experienced young drivers typically still in the provisional stage of graduated licensing systems.

Patient-specific computational biomechanics for simulating adolescent scoliosis surgery : predicted vs clinical correction for a preliminary series of six patients

Scoliosis is a spinal deformity that requires surgical correction in progressive cases. In order to optimize surgical outcomes, patient-specific finite element models are being developed by our group. In this paper, a single rod anterior correction procedure is simulated for a group of six scoliosis patients. For each patient, personalised model geometry was derived from low-dose CT scans, and clinically measured intra-operative corrective forces were applied. However, tissue material properties were not patient-specific, being derived from existing literature. Clinically, the patient group had a mean initial Cobb angle of 47.3 degrees, which was corrected to 17.5 degrees after surgery. The mean simulated post-operative Cobb angle for the group was 18.1 degrees. Although this represents good agreement between clinical and simulated corrections, the discrepancy between clinical and simulated Cobb angle for individual patients varied between -10.3 and +8.6 degrees, with only three of the six patients matching the clinical result to within accepted Cobb measurement error of +-5 degrees. The results of this study suggest that spinal tissue material properties play an important role in governing the correction obtained during surgery, and that patient-specific modelling approaches must address the question of how to prescribe patient-specific soft tissue properties for spine surgery simulation.

Linking the American Time Use Survey (ATUS) and the Compendium of Physical Activities : methods and rationale

Background: The 2003 Bureau of Labor Statistics American Time Use Survey (ATUS) contains 438 distinct primary activity variables that can be analyzed with regard to how time is spent by Americans. The Compendium of Physical Activities is used to code physical activities derived from various surveys, logs, diaries, etc to facilitate comparison of coded intensity levels across studies. ------ ----- Methods: This paper describes the methods, challenges, and rationale for linking Compendium estimates of physical activity intensity (METs, metabolic equivalents) with all activities reported in the 2003 ATUS. ----- ----- Results: The assigned ATUS intensity levels are not intended to compute the energy costs of physical activity in individuals. Instead, they are intended to be used to identify time spent in activities broadly classified by type and intensity. This function will complement public health surveillance systems and aid in policy and health-promotion activities. For example, at least one of the future projects of this process is the descriptive epidemiology of time spent in common physical activity intensity categories. ----- ----- Conclusions: The process of metabolic coding of the ATUS by linking it with the Compendium of Physical Activities can make important contributions to our understanding of Americans’ time spent in health-related physical activity.

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'.

Biaxial cell stimulation : a mechanical validation

To analyse mechanotransduction resulting from tensile loading under defined conditions, various devices for in vitro cell stimulation have been developed. This work aimed to determine the strain distribution on the membrane of a commercially available device and its consistency with rising cycle numbers, as well as the amount of strain transferred to adherent cells. The strains and their behaviour within the stimulation device were determined using digital image correlation (DIC). The strain transferred to cells was measured on eGFP-transfected bone marrow-derived cells imaged with a fluorescence microscope. The analysis was performed by determining the coordinates of prominent positions on the cells, calculating vectors between the coordinates and their length changes with increasing applied tensile strain. The stimulation device was found to apply homogeneous (mean of standard deviations approx. 2% of mean strain) and reproducible strains in the central well area. However, on average, only half of the applied strain was transferred to the bone marrow-derived cells. Furthermore, the strain measured within the device increased significantly with an increasing number of cycles while the membrane's Young's modulus decreased, indicating permanent changes in the material during extended use. Thus, strain magnitudes do not match the system readout and results require careful interpretation, especially at high cycle numbers.

A mathematical model of chlamydial infection incorporating movement of chlamydial particles

We present a spatiotemporal mathematical model of chlamydial infection, host immune response and spatial movement of infectious particles. The re- sulting partial differential equations model both the dynamics of the infection and changes in infection profile observed spatially along the length of the host genital tract. This model advances previous chlamydia modelling by incorporating spatial change, which we also demonstrate to be essential when the timescale for movement of infectious particles is equal to, or shorter than, the developmental cycle timescale. Numerical solutions and model analysis are carried out, and we present a hypothesis regarding the potential for treatment and prevention of infection by increasing chlamydial particle motility.

Ghrelin and cancer

Ghrelin is a peptide hormone that was originally isolated from the stomach as the endogenous ligand for the growth hormone secretagogue receptor (GHSR). Ghrelin has many functions, including the regulation of appetite and gut motility, growth hormone release from the anterior pituitary and roles in the cardiovascular and immune systems. Ghrelin and its receptor are expressed in a number of cancers and cancer cell lines and may play a role in processes associated with cancer progression, including cell proliferation, apoptosis, and cell invasion and migration.

Preliminary characterisation of the surface of cartilage following exposure to saturated and unsaturated synthetic lipids

Articular cartilage is covered by a microscopic structure known as surface amorphous layer. This surface structure is often the first victim of attack during cartilage degeneration, thereby resulting in a gross impairment in cartilage function such as lubrication and load bearing. We hypothesize that incubation of degraded cartilage in solutions of different species of synthetic surface active phospholipids (saturated and unsaturated species) can remodel this lost surface structure. To test this hypothesis, the structural configuration of the surface of articular cartilage was studied and characterised with the lipid filled surface amorphous layer intact using the AFM. The results were then compared with those obtained following a systematic removal (delipidization) and replacement (relipidization) of this layer. Our results show that the unsaturated surfactant partially restored the lost surface amorphous layer while the saturated surfactant specie settled on the surface due to its poor solubility in aqueous solution.