Sustained release and osteogenic potential of heparan sulfate-doped fibrin glue scaffolds within a rat cranial model

This paper explores the potential therapeutic role of the naturally occurring sugar heparan sulfate (HS) for the augmentation of bone repair. Scaffolds comprising fibrin glue loaded with 5 lg of embryonically derived HS were assessed, firstly as a release-reservoir, and secondly as a scaffold to stimulate bone regeneration in a critical size rat cranial defect. We show HS-loaded scaffolds have a uniform distribution of HS, which was readily released with a typical burst phase, quickly followed by a prolonged delivery lasting several days. Importantly, the released HS contributed to improved wound healing over a 3-month period as determined by microcomputed tomography (lCT) scanning, histology, histomorphometry, and PCR for osteogenic markers. In all cases, only minimal healing was observed after 1 and 3 months in the absence of HS. In contrast, marked healing was observed by 3 months following HS treatment, with nearly full closure of the defect site. PCR analysis showed significant increases in the gene expression of the osteogenic markers Runx2, alkaline phosphatase, and osteopontin in the heparin sulfate group compared with controls. These results further emphasize the important role HS plays in augmenting wound healing, and its successful delivery in a hydrogel provides a novel alternative to autologous bone graft and growth factorbased therapies.

Autocrine fibroblast growth factor 2 increases the multipotentiality of human adipose-derived mesenchymal stem cells

Multipotent mesenchymal stem cells (MSCs), first identified in the bone marrow, have subsequently been found in many other tissues, including fat, cartilage, muscle, and bone. Adipose tissue has been identified as an alternative to bone marrow as a source for the isolation of MSCs, as it is neither limited in volume nor as invasive in the harvesting. This study compares the multipotentiality of bone marrow-derived mesenchymal stem cells (BMSCs) with that of adipose-derived mesenchymal stem cells (AMSCs) from 12 age- and sex-matched donors. Phenotypically, the cells are very similar, with only three surface markers, CD106, CD146, and HLA-ABC, differentially expressed in the BMSCs. Although colony-forming units-fibroblastic numbers in BMSCs were higher than in AMSCs, the expression of multiple stem cell-related genes, like that of fibroblast growth factor 2 (FGF2), the Wnt pathway effectors FRAT1 and frizzled 1, and other self-renewal markers, was greater in AMSCs. Furthermore, AMSCs displayed enhanced osteogenic and adipogenic potential, whereas BMSCs formed chondrocytes more readily than AMSCs. However, by removing the effects of proliferation from the experiment, AMSCs no longer out-performed BMSCs in their ability to undergo osteogenic and adipogenic differentiation. Inhibition of the FGF2/fibroblast growth factor receptor 1 signaling pathway demonstrated that FGF2 is required for the proliferation of both AMSCs and BMSCs, yet blocking FGF2 signaling had no direct effect on osteogenic differentiation. Disclosure of potential conflicts of interest is found at the end of this article.

The influence of fibrin based hydrogels on the chondrogenic differentiation of human bone marrow stromal cells

Mesenchymal Stem Cells (MSC) are frequently incorporated into osteochondral implants and cell seeding is often facilitated with hydrogels which exert a profound influence on the chondrogenic differentiation of MSC. An attempt was made to elucidate this effect by comparing the chondrogenic differentiation of Bone Marrow Stromal Cells (BMSC) in fibrin and fibrin alginate composites. A biphasic osteochondral model which simulated the native in vivo environment was employed in the study. In the first stage of the experiment, BMSC was encapsulated in fibrin, Fibrin Alginate 0.3% (FA0.3) and 0.6% (FA0.6). Chondrogenic differentiation within these cell-hydrogel pellets was compared against that of standard cell pellets under inductive conditions and the matrices which supported chondrogenesis were used in the cartilage phase of biphasic constructs. Neo-cartilage growth was monitored in these cocultures. It was observed that hydrogel encapsulation influenced mesenchymal condensation which preceded chondrogenic differentiation. Early cell agglomeration was observed in fibrin as compared to fibrin alginate composites. These fibrin encapsulated cells differentiated into chondrocytes which secreted aggrecan and collagen II. When the alginate content rose from 0.3 to 0.6%, chondrogenic differentiation declined with a reduction in the expression of collagen II and aggrecan. Fibrin and FA0.3 were tested in the cartilage phase of the biphasic osteochondral constructs and the former supported superior cartilage growth with higher cellularity, total Glycosaminoglycan (GAG) and collagen II levels. The FA0.3 cartilage phase was found to be fragmented and partially calcified. The use of fibrin for cartilage repair was advocated as it facilitated BMSC chondrogenesis and cartilaginous growth in an osteochondral environment.

Differences between in vitro viability and differentiation and in vivo bone-forming efficacy of human mesenchymal stem cells cultured on PCL-TCP scaffolds

Human mesenchymal stem cells (hMSCs) possess great therapeutic potential for the treatment of bone disease and fracture non-union. Too often however, in vitro evidence alone of the interaction between hMSCs and the biomaterial of choice is used as justification for continued development of the material into the clinic. Clearly for hMSC-based regenerative medicine to be successful for the treatment of orthopaedic trauma, it is crucial to transplant hMSCs with a suitable carrier that facilitates their survival, optimal proliferation and osteogenic differentiation in vitro and in vivo. This motivated us to evaluate the use of polycaprolactone-20% tricalcium phosphate (PCL-TCP) scaffolds produced by fused deposition modeling for the delivery of hMSCs. When hMSCs were cultured on the PCL-TCP scaffolds and imaged by a combination of phase contrast, scanning electron and confocal laser microscopy, we observed five distinct stages of colonization over a 21-day period that were characterized by cell attachment, spreading, cellular bridging, the formation of a dense cellular mass and the accumulation of a mineralized extracellular matrix when induced with osteogenic stimulants. Having established that PCL-TCP scaffolds are able to support hMSC proliferation and osteogenic differentiation, we next tested the in vivo efficacy of hMSC-loaded PCL-TCP scaffolds in nude rat critical-sized femoral defects. We found that fluorescently labeled hMSCs survived in the defect site for up to 3 weeks post-transplantation. However, only 50% of the femoral defects treated with hMSCs responded favorably as determined by new bone volume. As such, we show that verification of hMSC viability and differentiation in vitro is not sufficient to predict the efficacy of transplanted stem cells to consistently promote bone formation in orthotopic defects in vivo.

Flower animals

The coral reefs around the world may be likened to canaries down the mineshaft of global warming. These sensitive plant-like animals have evolved for life in tropical seas. Their needs are quite specific – not too cold, not too hot. A rise of as little as one degree Celsius is enough to cause some bleaching of these colourful jewels of the sea. Many climate models indicate we can expect sea temperature increases of between two and six degrees Celsius. Research - such as that detailed in a 2004 report by the University of Queensland’s Centre for Marine Studies – indicates that by the year 2050 most of the worlds major reef systems will be dead. Many of us have heard this kind of information, but it remains difficult to comprehend. It’s almost impossible to imagine the death of the Great Barrier Reef. Some six to nine thousand years old and visible from space, it is the world’s largest structure created by living organisms. Yet whilst it is hard to believe, this gentle, sensitive giant is at grave risk because it cannot adapt quickly enough to the changes in the environment. This cluster of fluffy felt brain coral sculptures are connected in real time to temperature data collected by monitoring stations within the Great Barrier Reef, that form part of the Australian Institute of Marine Science’s Great Barrier Reed Ocean Observing System. These corals display illumination patterns showing changes in sea temperature at Heron Reef, one of the 2,900 reefs that comprise the Great Barrier Reef. Their spectrum of colour ranges from cool hues, through warm tones to bright white when temperatures exceed those that tropical corals are able to tolerate over sustained periods. The Flower Animals also blush in colour and make sound when people come within close proximity. In a reef, fishes and other creatures generate significant amounts of sound. These cacophonies are considered an indicator of reef health, and are used by reef fish to determine where they can best live and forage.

AITPM national newsletter November 2008 president's report

As we’re moving toward the end of the year, it’s not hard to notice everyone starting to rush a bit more. So much so, that in some cases people can lose their cool when they’re out and about. Recently I viewed an episode of Jenny Brockie’s Insight program on SBS on the topic of “rage”. The program covered many areas of life, but it highlighted the issue of rage against taxi drivers in Melbourne and showed some archival footage of the recent taxi drivers’ protest on the issue, next to Flinders Street Station. Serendipitously, perhaps, I picked up Brisbane’s City News as I was eating lunch in town a few days later, and there was an article on Brisbane taxi stand supervisors, citing that some feared to go to work on Friday and Saturday nights as they were not infrequently assaulted by drunken revellers waiting in the long queues for their taxi ride home.

Heat strain, hydration status, and symptoms of heat illness in surface mine workers

The aim of the research program was to evaluate the heat strain, hydration status, and heat illness symptoms experienced by surface mine workers. An initial investigation involved 91 surface miners completing a heat stress questionnaire; assessing the work environment, hydration practices, and heat illness symptom experience. The key findings included 1) more than 80 % of workers experienced at least one symptom of heat illness over a 12 month period; and 2) the risk of moderate symptoms of heat illness increased with the severity of dehydration. These findings highlight a health and safety concern for surface miners, as experiencing symptoms of heat illness is an indication that the physiological systems of the body may be struggling to meet the demands of thermoregulation. To illuminate these findings a field investigation to monitor the heat strain and hydration status of surface miners was proposed. Two preliminary studies were conducted to ensure accurate and reliable data collection techniques. Firstly, a study was undertaken to determine a calibration procedure to ensure the accuracy of core body temperature measurement via an ingestible sensor. A water bath was heated to several temperatures between 23 . 51 ¢ªC, allowing for comparison of the temperature recorded by the sensors and a traceable thermometer. A positive systematic bias was observed and indicated a need for calibration. It was concluded that a linear regression should be developed for each sensor prior to ingestion, allowing for a correction to be applied to the raw data. Secondly, hydration status was to be assessed through urine specific gravity measurement. It was foreseeable that practical limitations on mine sites would delay the time between urine collection and analysis. A study was undertaken to assess the reliability of urine analysis over time. Measurement of urine specific gravity was found to be reliable up to 24 hours post urine collection and was suitable to be used in the field study. Twenty-nine surface miners (14 drillers [winter] and 15 blast crew [summer]) were monitored during a normal work shift. Core body temperature was recorded continuously. Average mean core body temperature was 37.5 and 37.4 ¢ªC for blast crew and drillers, with average maximum body temperatures of 38.0 and 37.9 ¢ªC respectively. The highest body temperature recorded was 38.4 ¢ªC. Urine samples were collected at each void for specific gravity measurement. The average mean urine specific gravity was 1.024 and 1.021 for blast crew and drillers respectively. The Heat Illness Symptoms Index was used to evaluate the experience of heat illness symptoms on shift. Over 70 % of drillers and over 80 % of blast crew reported at least one symptom. It was concluded that 1) heat strain remained within the recommended limits for acclimatised workers; and 2) the majority of workers were dehydrated before commencing their shift, and tend to remain dehydrated for the duration. Dehydration was identified as the primary issue for surface miners working in the heat. Therefore continued study focused on investigating a novel approach to monitoring hydration status. The final aim of this research program was to investigate the influence dehydration has on intraocular pressure (IOP); and subsequently, whether IOP could provide a novel indicator of hydration status. Seven males completed 90 minutes of walking in both a cool and hot climate with fluid restriction. Hydration variables and intraocular pressure were measured at baseline and at 30 minute intervals. Participants became dehydrated during the trial in the heat but maintained hydration status in the cool. Intraocular pressure progressively declined in the trial in the heat but remained relatively stable when hydration was maintained. A significant relationship was observed between intraocular pressure and both body mass loss and plasma osmolality. This evidence suggests that intraocular pressure is influenced by changes in hydration status. Further research is required to determine if intraocular pressure could be utilised as an indirect indicator of hydration status.

Glycosaminoglycan and growth factor mediated murine calvarial cell proliferation

Understanding the complex mechanisms underlying bone remodeling is crucial to the development of novel therapeutics. Glycosaminoglycans (GAGs) localised to the extracellular matrix (ECM) of bone are thought to play a key role in mediating aspects of bone development. The influence of isolated GAGs was studied by utilising in vitro murine calvarial monolayer and organ culture model systems. Addition of GAG preparations extracted from the cell surface of human osteoblasts at high concentrations (5 microg/ml) resulted in decreased proliferation of cells and decreased suture width and number of bone lining cells in calvarial sections. When we investigated potential interactions between the growth factors fibroblast growth factor-2 (FGF2), bone morphogenic protein-2 (BMP2) and transforming growth factor-beta1 (TGFbeta1) and the isolated cell surface GAGs, differences between the two model systems emerged. The cell culture system demonstrated a potentiating role for the isolated GAGs in the inhibition of FGF2 and TGFbeta1 actions. In contrast, the organ culture system demonstrated an enhanced stimulation of TFGbeta1 effects. These results emphasise the role of the ECM in mediating the interactions between GAGs and growth factors during bone development and suggest the GAG preparations contain potent inhibitory or stimulatory components able to mediate growth factor activity.

Purification and characterization of heparan sulfate from human primary osteoblasts

Heparan sulfate (HS) is a linear, highly variable, highly sulfated glycosaminoglycan sugar whose biological activity largely depends on internal sulfated domains that mediate specific binding to an extensive range of proteins. In this study we employed anion exchange chromatography, molecular sieving and enzymatic cleavage on HS fractions purified from three compartments of cultured osteoblasts-soluble conditioned media, cell surface, and extracellular matrix (ECM). We demonstrate that the composition of HS chains purified from the different compartments is structurally non-identical by a number of parameters, and that these differences have significant ramifications for their ligand-binding properties. The HS chains purified of conditioned medium had twice the binding affinity for FGF2 when compared with either cell surface or ECM HS. In contrast, similar binding of BMP2 to the three types of HS was observed. These results suggest that different biological compartments of cultured cells have structurally and functionally distinct HS species that help to modulate the flow of HS-dependent factors between the ECM and the cell surface.

Sensitivity of building zones to potential global warming

As global warming entails new conditions for the built environment, the thermal and energy performance of existing buildings, which are designed based on current weather data, may become unclear and remain a great concern. Through building computer simulation and qualitative analysis of the weighted factor for the outdoor temperature impact on building energy and thermal performance, this paper investigates the sensitivity of different office building zoning to the potential global warming. A standard office building type is examined for all eight capital cities in Australia. Results show that comparing the middle and top floors, except for cool climate (i.e. Hobart), the ground floor appears to be the most sensitive to the effect of global warming and has the highest tendency for a overheating problem. From the analysis of the responses of different zone orientations to the outdoor air temperature increase, it is also found that there are widely varied responses between zone orientations, with South zone (in the southern hemisphere) being the most sensitive. With an increased external air temperature, the variation between different floors or zone orientations will become more significant, up to 53 percent increase of overheating hours in Darwin and 47 percent increase of cooling load in Hobart.