Application of hyperbaric oxygen in bone tissue engineering : effect of hyperbaric oxygen treatment on bone marrow stem cells

and non-union of bony fractures has been proposed since 1966, little has been known about the effect of HBOT on bone marrow stem cells (BMSC). The aim of this study is to investigate the effect of HBO treatment on osteogenetic differentiation of BMSC and potential application in bone tissue engineering. Adhesive stromal cells harvested from bone marrow were characterized by mesenchymal differentiation potential, cell surface markers and their proliferation capacity. Mesenchymal stem cells, which demonstrated osteogenic, chondrogenic and adipogenic differentiation potential and expressed positively for CD 29, CD 44, CD 73, CD 90, CD 105, CD 166 and negatively for CD34 and CD 45, were selected and treated in a laboratory-scale HBO chamber using different oxygen pressures and exposure times. No obvious effect of HBO treatment on BMSC proliferation was noticed. However, cytotoxic effects of HBO were considerably less pronounced when cells were cultured in medium supplemented with 10% FBS in comparison to medium supplemented with 2% FCS, as was evaluated by WST-1 assay. Under HBO treatment, bone nodules were formed in three days, which was clearly revealed by Von Kossa staining. In contrasts, without HBO treatment, bone nodules were not detected until 9-12 days using the same inducing culture media. Calcium deposition was also significantly increased after three days of HBO treatments compared to no HBO treatment. In addition it was also found that oxygen played a direct role in the enhancement of BMSC osteogenic differentiation, which was independent of the effect of air pressure.

Inhibition of integrative cartilage repair by proteoglycan 4 in synovial fluid

To determine the effects of the articular cartilage surface, as well as synovial fluid (SF) and its components, specifically proteoglycan 4 (PRG4) and hyaluronic acid (HA), on integrative cartilage repair in vitro. Methods. Blocks of calf articular cartilage were harvested, some with the articular surface intact and others without. Some of the latter types of blocks were pretreated with trypsin, and then with bovine serum albumin, SF, PRG4, or HA. Immunolocalization of PRG4 on cartilage surfaces was performed after treatment. Pairs of similarly treated cartilage blocks were incubated in partial apposition for 2 weeks in medium supplemented with serum and 3 H-proline. Following culture, mechanical integration between apposed cartilage blocks was assessed by measuring adhesive strength, and protein biosynthesis and deposition were determined by incorporated 3 H-proline. Results. Samples with articular surfaces in apposition exhibited little integrative repair compared with samples with cut surfaces in apposition. PRG4 was immunolocalized at the articular cartilage surface, but not in deeper, cut surfaces (without treatment). Cartilage samples treated with trypsin and then with SF or PRG4 exhibited an inhibition of integrative repair and positive immunostaining for PRG4 at treated surfaces compared with normal cut cartilage samples, while samples treated with HA exhibited neither inhibited integrative repair nor PRG4 at the tissue surfaces. Deposition of newly synthesized protein was relatively similar under conditions in which integration differed significantly. Conclusion. These results support the concept that PRG4 in SF, which normally contributes to cartilage lubrication, can inhibit integrative cartilage repair. This has the desirable effect of preventing fusion of apposing surfaces of articulating cartilage, but has the undesirable effect of inhibiting integrative repair.

An approach for current balancing in distribution networks with rooftop PVs

Current unbalance is a significant power quality problem in distribution networks. This problem increases further with the increased penetration of single-phase photovoltaic cells. In this paper, a new approach is developed for current unbalance reduction in medium voltage distribution networks. The method is based on utilization of three single-phase voltage source converters connected in delta configuration between the phases. Each converter is controlled to function as a varying capacitor. The combination of the load and the compensator will result in a balanced load with unity power factor. The efficacy of the proposed current unbalance reduction concept is verified through dynamic simulations in PSCAD/EMTDC.

Intrinsics and dynamics of fat grafts : an in vitro study

Background Despite a revived interest in fat grafting procedures, clinicians still fail to demonstrate clearly the in vivo behavior of fat grafts as a dynamic tissue substitute. However, the basic principles in cellular biology teach us that cells can survive and develop, provided that a structural matrix exists that directs their behavior. The purpose of this in vitro study was to analyze that behavior of crude fat grafts, cultured on a three-dimensional laminin-rich matrix. Methods Nonprocessed, human fat biopsy specimens (approximately 1 mm) were inoculated on Matrigel-coated wells to which culture medium was added. The control group consisted of fat biopsy specimens embedded in medium alone. The cellular proliferation pattern was followed over 6 weeks. Additional cultures of primary generated cellular spheroids were performed and eventually subjected to adipogenic differentiation media. Results A progressive outgrowth of fibroblast-like cells from the core fat biopsy specimen was observed in both groups. Within the Matrigel group, an interconnecting three-dimensional network of spindle-shaped cells was established. This new cell colony reproduced spheroids that functioned again as solitary sources of cellular proliferation. Addition of differentiation media resulted in lipid droplet deposition in the majority of generated cells, indicating the initial steps of adipogenic differentiation. Conclusions The authors noticed that crude, nonprocessed fat biopsy specimens do have considerable potential for future tissue engineering-based applications, provided that the basic principles of developmental, cellular biology are respected. Spontaneous in vitro expansion of the stromal cells present in fat grafts within autologous and injectable matrices could create "off-the-shelf" therapies for reconstructive procedures.

Localization of fibronectin in the rat testis : effects of serum and dibutyryl cyclic AMP on fibronectin deposition by peritubular myoid cells in culture

The peritubular zone of the rat testis has an extensive extracellular matrix (ECM). Fibronectin (FN) is distributed primarily in the basal lamina of the seminiferous tubule boundary tissue and is synthesized by peritubular myoid cells. Several extracellular changes are mediated by growth factors and these changes occur at the time of hormone mediated testicular development, particularly in the peritubular zone. The effects of serum or dibutyryl cyclic AMP (cAMP) on FN production by the mesenchymal peritubular myoid cells were evaluated. Rats of various ages (10, 15, 20, 40 and 80 days) were employed for immunofluorescent localization of rat testicular FN in frozen sections. In all age groups tested, FN was primarily present in a broad layer around each seminiferous tubule, and blood vessel, and in variable distribution throughout the interstitial stroma. By day 20 there was no clear distinction in FN staining between the peritubular zone and the interstitial tissue. This indicates an involvement of FN in the ECM developments which occur in the peritubular zone of the testis at this time. The peritubular myoid cells were isolated from 20-22 day old rat testis and cultured on glass coverslips. These cells were grown to confluence with 10% fetal calf serum (FCS) in medium until day 4 and then subcultured to have secondary monocultures maintained with or without serum. By means of immunofluorescence and cytochemistry using avidin-biotin peroxidase complex it was observed that peritubular myoid cells were positive for FN and most of the FN was localized in the perinuclear region. Subcultured peritubular myoid cells maintained for 4 days in medium containing FCS developed an extensive interconnecting FN matrix. In the presence of 0.5 mM cAMP in culture, FN became localized along the filamentous process of peritubular myoid cells and more prominently in the areas of triangulated multi-cell aggregates as well as on the surface of the contracted small spherical cells. The addition of cAMP in the presence of FCS, also caused a noticeable change in the staining pattern; FN was detected along the filamentous process developing into a complex network of cells encased in an extensive matrix. It would appear that the translocation of FN in the cytoplasmic extensions of peritubular myoid cells may be a direct consequence of morphological changes associated with metabolic regulation of cAMP. This may also be related to the puberty associated development of in vivo changes in the ECM produced by peritubular myoid cells.

Impact response and parametric studies of reinforced concrete circular columns

This paper presents a detailed description of the influence of critical parameters that govern the vulnerability of columns under lateral impact loads. Numerical simulations are conducted by using the Finite Element program LS-DYNA, incorporating steel reinforcement, material models and strain rate effects. A simplified method based on impact pulse generated from full scale impact tests is used for impact reconstruction and effects of the various pulse loading parameters are investigated under low to medium velocity impacts. A constitutive material model which can simulate failures under tri-axial state of stresses is used for concrete. Confinement effects are also introduced to the numerical simulation and columns of Grade 30 to 50 concrete under pure axial loading are analysed in detail. This research confirmed that the vulnerability of the axially loaded columns can be mitigated by reducing the slenderness ratio and concrete grade, and by choosing the design option with a minimal amount of longitudinal steel. Additionally, it is evident that approximately a 50% increase in impact capacity can be gained for columns in medium rise buildings by enhancing the confinement effects alone. Results also indicated that the ductility as well as the mode of failure under impact can be changed with the volumetric ratio of lateral steel. Moreover, to increase the impact capacity of the vulnerable columns, a higher confining stress is required. The general provisions of current design codes do not sufficiently cover this aspect and hence this research will provide additional guidelines to overcome the inadequacies of code provisions.

A comparative study : regulatory requirements on subdivisions for increasing passive cooling

A major project in the Sustainable Built Assets core area is the Sustainable Sub-divisions – Ventilation Project that is the second stage of a planned series of research projects focusing on sustainable sub-divisions. The initial project, Sustainable Sub-divisions: Energy focused on energy efficiency and examined the link between dwelling energy efficiency and sub-divisional layout. In addition, the potential for on site electricity generation, especially in medium and high-density developments, was also examined. That project recommended that an existing lot-rating methodology be adapted for use in SEQ through the inclusion of sub divisional appropriate ventilation data. Acquiring that data is the object of this project. The Sustainable Sub-divisions; Ventilation Project will produce a series of reports. The first report (Report 2002-077-B-01) summarised the results from an industry workshop and interviews that were conducted to ascertain the current attitudes and methodologies used in contemporary sub-division design in South East Queensland. The second report (Report 2002-077-B-02) described how the project is being delivered as outlined in the Project Agreement. It included the selection of the case study dwellings and monitoring equipment and data management process. This third report (Report 2002-077-B-03) provides an analysis and review of the approaches recommended by leading experts, government bodies and professional organizations throughout Australia that aim to increase the potential for passive cooling and heating at the subdivision stage. This data will inform issues discussed on the development of the enhanced lot-rating methodology in other reports of this series. The final report, due in June 2007, will detail the analysis of data for winter 2006 and summer 2007, leading to the development and delivery of the enhanced lot-rating methodology.

A new multilevel converter configuration for high power and high quality applications

The Queensland University of Technology (QUT) allows the presentation of theses for the Degree of Doctor of Philosophy in the format of published or submitted papers, where such papers have been published, accepted or submitted during the period of candidature. This thesis is composed of ten published /submitted papers and book chapters of which nine have been published and one is under review. This project is financially supported by an Australian Research Council (ARC) Discovery Grant with the aim of investigating multilevel topologies for high quality and high power applications, with specific emphasis on renewable energy systems. The rapid evolution of renewable energy within the last several years has resulted in the design of efficient power converters suitable for medium and high-power applications such as wind turbine and photovoltaic (PV) systems. Today, the industrial trend is moving away from heavy and bulky passive components to power converter systems that use more and more semiconductor elements controlled by powerful processor systems. However, it is hard to connect the traditional converters to the high and medium voltage grids, as a single power switch cannot stand at high voltage. For these reasons, a new family of multilevel inverters has appeared as a solution for working with higher voltage levels. Besides this important feature, multilevel converters have the capability to generate stepped waveforms. Consequently, in comparison with conventional two-level inverters, they present lower switching losses, lower voltage stress across loads, lower electromagnetic interference (EMI) and higher quality output waveforms. These properties enable the connection of renewable energy sources directly to the grid without using expensive, bulky, heavy line transformers. Additionally, they minimize the size of the passive filter and increase the durability of electrical devices. However, multilevel converters have only been utilised in very particular applications, mainly due to the structural limitations, high cost and complexity of the multilevel converter system and control. New developments in the fields of power semiconductor switches and processors will favor the multilevel converters for many other fields of application. The main application for the multilevel converter presented in this work is the front-end power converter in renewable energy systems. Diode-clamped and cascade converters are the most common type of multilevel converters widely used in different renewable energy system applications. However, some drawbacks – such as capacitor voltage imbalance, number of components, and complexity of the control system – still exist, and these are investigated in the framework of this thesis. Various simulations using software simulation tools are undertaken and are used to study different cases. The feasibility of the developments is underlined with a series of experimental results. This thesis is divided into two main sections. The first section focuses on solving the capacitor voltage imbalance for a wide range of applications, and on decreasing the complexity of the control strategy on the inverter side. The idea of using sharing switches at the output structure of the DC-DC front-end converters is proposed to balance the series DC link capacitors. A new family of multioutput DC-DC converters is proposed for renewable energy systems connected to the DC link voltage of diode-clamped converters. The main objective of this type of converter is the sharing of the total output voltage into several series voltage levels using sharing switches. This solves the problems associated with capacitor voltage imbalance in diode-clamped multilevel converters. These converters adjust the variable and unregulated DC voltage generated by renewable energy systems (such as PV) to the desirable series multiple voltage levels at the inverter DC side. A multi-output boost (MOB) converter, with one inductor and series output voltage, is presented. This converter is suitable for renewable energy systems based on diode-clamped converters because it boosts the low output voltage and provides the series capacitor at the output side. A simple control strategy using cross voltage control with internal current loop is presented to obtain the desired voltage levels at the output voltage. The proposed topology and control strategy are validated by simulation and hardware results. Using the idea of voltage sharing switches, the circuit structure of different topologies of multi-output DC-DC converters – or multi-output voltage sharing (MOVS) converters – have been proposed. In order to verify the feasibility of this topology and its application, steady state and dynamic analyses have been carried out. Simulation and experiments using the proposed control strategy have verified the mathematical analysis. The second part of this thesis addresses the second problem of multilevel converters: the need to improve their quality with minimum cost and complexity. This is related to utilising asymmetrical multilevel topologies instead of conventional multilevel converters; this can increase the quality of output waveforms with a minimum number of components. It also allows for a reduction in the cost and complexity of systems while maintaining the same output quality, or for an increase in the quality while maintaining the same cost and complexity. Therefore, the asymmetrical configuration for two common types of multilevel converters – diode-clamped and cascade converters – is investigated. Also, as well as addressing the maximisation of the output voltage resolution, some technical issues – such as adjacent switching vectors – should be taken into account in asymmetrical multilevel configurations to keep the total harmonic distortion (THD) and switching losses to a minimum. Thus, the asymmetrical diode-clamped converter is proposed. An appropriate asymmetrical DC link arrangement is presented for four-level diode-clamped converters by keeping adjacent switching vectors. In this way, five-level inverter performance is achieved for the same level of complexity of the four-level inverter. Dealing with the capacitor voltage imbalance problem in asymmetrical diodeclamped converters has inspired the proposal for two different DC-DC topologies with a suitable control strategy. A Triple-Output Boost (TOB) converter and a Boost 3-Output Voltage Sharing (Boost-3OVS) converter connected to the four-level diode-clamped converter are proposed to arrange the proposed asymmetrical DC link for the high modulation indices and unity power factor. Cascade converters have shown their abilities and strengths in medium and high power applications. Using asymmetrical H-bridge inverters, more voltage levels can be generated in output voltage with the same number of components as the symmetrical converters. The concept of cascading multilevel H-bridge cells is used to propose a fifteen-level cascade inverter using a four-level H-bridge symmetrical diode-clamped converter, cascaded with classical two-level Hbridge inverters. A DC voltage ratio of cells is presented to obtain maximum voltage levels on output voltage, with adjacent switching vectors between all possible voltage levels; this can minimize the switching losses. This structure can save five isolated DC sources and twelve switches in comparison to conventional cascade converters with series two-level H bridge inverters. To increase the quality in presented hybrid topology with minimum number of components, a new cascade inverter is verified by cascading an asymmetrical four-level H-bridge diode-clamped inverter. An inverter with nineteen-level performance was achieved. This synthesizes more voltage levels with lower voltage and current THD, rather than using a symmetrical diode-clamped inverter with the same configuration and equivalent number of power components. Two different predictive current control methods for the switching states selection are proposed to minimise either losses or THD of voltage in hybrid converters. High voltage spikes at switching time in experimental results and investigation of a diode-clamped inverter structure raised another problem associated with high-level high voltage multilevel converters. Power switching components with fast switching, combined with hard switched-converters, produce high di/dt during turn off time. Thus, stray inductance of interconnections becomes an important issue and raises overvoltage and EMI issues correlated to the number of components. Planar busbar is a good candidate to reduce interconnection inductance in high power inverters compared with cables. The effect of different transient current loops on busbar physical structure of the high-voltage highlevel diode-clamped converters is highlighted. Design considerations of proper planar busbar are also presented to optimise the overall design of diode-clamped converters.

Compression for preventing recurrence of venous ulcers

Background: Up to 1% of adults will suffer from leg ulceration at some time. The majority of leg ulcers are venous in origin and are caused by high pressure in the veins due to blockage or weakness of the valves in the veins of the leg. Prevention and treatment of venous ulcers is aimed at reducing the pressure either by removing / repairing the veins, or by applying compression bandages / stockings to reduce the pressure in the veins. The vast majority of venous ulcers are healed using compression bandages. Once healed they often recur and so it is customary to continue applying compression in the form of bandages, tights, stockings or socks in order to prevent recurrence. Compression bandages or hosiery (tights, stockings, socks) are often applied for ulcer prevention. Objectives To assess the effects of compression hosiery (socks, stockings, tights) or bandages in preventing the recurrence of venous ulcers. To determine whether there is an optimum pressure/type of compression to prevent recurrence of venous ulcers. Search methods The searches for the review were first undertaken in 2000. For this update we searched the Cochrane Wounds Group Specialised Register (October 2007), The Cochrane Central Register of Controlled Trials (CENTRAL) - The Cochrane Library 2007 Issue 3, Ovid MEDLINE - 1950 to September Week 4 2007, Ovid EMBASE - 1980 to 2007 Week 40 and Ovid CINAHL - 1982 to October Week 1 2007. Selection criteria Randomised controlled trials evaluating compression bandages or hosiery for preventing venous leg ulcers. Data collection and analysis Data extraction and assessment of study quality were undertaken by two authors independently. Results No trials compared recurrence rates with and without compression. One trial (300 patients) compared high (UK Class 3) compression hosiery with moderate (UK Class 2) compression hosiery. A intention to treat analysis found no significant reduction in recurrence at five years follow up associated with high compression hosiery compared with moderate compression hosiery (relative risk of recurrence 0.82, 95% confidence interval 0.61 to 1.12). This analysis would tend to underestimate the effectiveness of the high compression hosiery because a significant proportion of people changed from high compression to medium compression hosiery. Compliance rates were significantly higher with medium compression than with high compression hosiery. One trial (166 patients) found no statistically significant difference in recurrence between two types of medium (UK Class 2) compression hosiery (relative risk of recurrence with Medi was 0.74, 95% confidence interval 0.45 to 1.2). Both trials reported that not wearing compression hosiery was strongly associated with ulcer recurrence and this is circumstantial evidence that compression reduces ulcer recurrence. No trials were found which evaluated compression bandages for preventing ulcer recurrence. Authors' conclusions No trials compared compression with vs no compression for prevention of ulcer recurrence. Not wearing compression was associated with recurrence in both studies identified in this review. This is circumstantial evidence of the benefit of compression in reducing recurrence. Recurrence rates may be lower in high compression hosiery than in medium compression hosiery and therefore patients should be offered the strongest compression with which they can comply. Further trials are needed to determine the effectiveness of hosiery prescribed in other settings, i.e. in the UK community, in countries other than the UK.

Fixed failure rate ambiguity validation methods for GPS and compass

Ambiguity resolution plays a crucial role in real time kinematic GNSS positioning which gives centimetre precision positioning results if all the ambiguities in each epoch are correctly fixed to integers. However, the incorrectly fixed ambiguities can result in large positioning offset up to several meters without notice. Hence, ambiguity validation is essential to control the ambiguity resolution quality. Currently, the most popular ambiguity validation is ratio test. The criterion of ratio test is often empirically determined. Empirically determined criterion can be dangerous, because a fixed criterion cannot fit all scenarios and does not directly control the ambiguity resolution risk. In practice, depending on the underlying model strength, the ratio test criterion can be too conservative for some model and becomes too risky for others. A more rational test method is to determine the criterion according to the underlying model and user requirement. Miss-detected incorrect integers will lead to a hazardous result, which should be strictly controlled. In ambiguity resolution miss-detected rate is often known as failure rate. In this paper, a fixed failure rate ratio test method is presented and applied in analysis of GPS and Compass positioning scenarios. A fixed failure rate approach is derived from the integer aperture estimation theory, which is theoretically rigorous. The criteria table for ratio test is computed based on extensive data simulations in the approach. The real-time users can determine the ratio test criterion by looking up the criteria table. This method has been applied in medium distance GPS ambiguity resolution but multi-constellation and high dimensional scenarios haven't been discussed so far. In this paper, a general ambiguity validation model is derived based on hypothesis test theory, and fixed failure rate approach is introduced, especially the relationship between ratio test threshold and failure rate is examined. In the last, Factors that influence fixed failure rate approach ratio test threshold is discussed according to extensive data simulation. The result shows that fixed failure rate approach is a more reasonable ambiguity validation method with proper stochastic model.

Development of an ultra-thin fibroin membrane for RPE cell transplantation

Purpose: One of the challenges associated with cell-based therapies for repairing the retina is the development of suitable materials on which to grow and transplant retinal cells. Using the ARPE-19 cell line, we have previously demonstrated the feasibility of growing RPE-derived cells on membranes prepared from the silk protein fibroin. The present study was aimed at developing a porous, ultra-thin fibroin membrane that might better support development of apical-basal polarity in culture, and to extend this work to primary cultures of human RPE cells. Methods: Ultra-thin fibroin membranes were prepared using a highly polished casting table coated with Topas® (a cyclic olefin copolymer) and a 1:0.03 aqueous solution of fibroin and PEO (Mv 900 000 g/mol). Following drying, the membranes were water annealed to make them water-stable, washed in water to remove PEO, sterilised by treatment with 95% ethanol, and washed extensively in saline. Primary cultures containing human RPE cells were established from donor posterior eye cups and maintained in DMEM/F12 medium supplemented with 10% fetal bovine serum and antibiotics. First passage cultures were seeded onto fibroin membranes pre-coated with vitronectin and grown for 6 weeks in medium supplemented with 1% serum. Comparative cultures were established on porous 1.0 µm pore PET membrane (Millipore) and using ARPE-19 cells. Results: The fibroin membranes displayed an average thickness of 3 µm and contained numerous dimples/pore-like structures of up to 3-5 µm in diameter. The primary cultures predominantly contained pigmented epithelial cells, but mesenchymal cells (presumed fibroblasts) were also often present. Passaged cultures appeared to attach equally well to either fibroin or PET membranes. Over time cells on either material adopted a more cobblestoned morphology. Conclusions: Progress has been made towards developing a porous ultra-thin fibroin membrane that supports cultivation of RPE cells. Further studies are required to determine the degree of membrane permeability and RPE polarity.

Packed bed bioreactor for the isolation and expansion of placental-derived Mesenchymal Stromal Cells

Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs.