Mineralization capacity of Runx2/Cbfa1-genetically engineered fibroblasts is scaffold dependent

Development of tissue-engineered constructs for skeletal regeneration of large critical-sized defects requires the identification of a sustained mineralizing cell source and careful optimization of scaffold architecture and surface properties. We have recently reported that Runx2-genetically engineered primary dermal fibroblasts express a mineralizing phenotype in monolayer culture, highlighting their potential as an autologous osteoblastic cell source which can be easily obtained in large quantities. The objective of the present study was to evaluate the osteogenic potential of Runx2-expressing fibroblasts when cultured in vitro on three commercially available scaffolds with divergent properties: fused deposition-modeled polycaprolactone (PCL), gas-foamed polylactide-co-glycolide (PLGA), and fibrous collagen disks. We demonstrate that the mineralization capacity of Runx2-engineered fibroblasts is scaffold dependent, with collagen foams exhibiting ten-fold higher mineral volume compared to PCL and PLGA matrices. Constructs were differentially colonized by genetically modified fibroblasts, but scaffold-directed changes in DNA content did not correlate with trends in mineral deposition. Sustained expression of Runx2 upregulated osteoblastic gene expression relative to unmodified control cells, and the magnitude of this expression was modulated by scaffold properties. Histological analyses revealed that matrix mineralization co-localized with cellular distribution, which was confined to the periphery of fibrous collagen and PLGA sponges and around the circumference of PCL microfilaments. Finally, FTIR spectroscopy verified that mineral deposits within all Runx2-engineered scaffolds displayed the chemical signature characteristic of carbonate-containing, poorly crystalline hydroxyapatite. These results highlight the important effect of scaffold properties on the capacity of Runx2-expressing primary dermal fibroblasts to differentiate into a mineralizing osteoblastic phenotype for bone tissue engineering applications.

Properties of porous structures prepared by stereolithography using a polylactide resin

Rapid prototyping techniques such as stereolithography allow for building designed tissue engineering scaffolds with high accuracy. In this work, a stereolithography resin based on poly(D,L-lactide) was developed. Biodegradable scaffolds with varying porosity were built from this resin. The scaffolds were analysed by μCT-scanning and compression testing. The porous structures showed excellent mechanical properties in the range of trabecular bone.

Bio-inspired multifunctional metallic foams through the fusion of different biological solutions

Nature is a school for scientists and engineers. Inherent multiscale structures of biological materials exhibit multifunctional integration. In nature, the lotus, the water strider, and the flying bird evolved different and optimized biological solutions to survive. In this contribution, inspired by the optimized solutions from the lotus leaf with superhydrophobic self-cleaning, the water strider leg with durable and robust superhydrophobicity, and the lightweight bird bone with hollow structures, multifunctional metallic foams with multiscale structures are fabricated, demonstrating low adhesive superhydrophobic self-cleaning, striking loading capacity, and superior repellency towards different corrosive solutions. This approach provides an effective avenue to the development of water strider robots and other aquatic smart devices floating on water. Furthermore, the resultant multifunctional metallic foam can be used to construct an oil/water separation apparatus, exhibiting a high separation efficiency and long-term repeatability. The presented approach should provide a promising solution for the design and construction of other multifunctional metallic foams in a large scale for practical applications in the petro-chemical field. Optimized biological solutions continue to inspire and to provide design idea for the construction of multiscale structures with multifunctional integration. Inspired by the optimized biological solutions from the lotus leaf with superhydrophobic self-cleaning, the water strider leg with durable and robust superhydrophobicity, and the lightweight bird bone with hollow structures, multifunctional metallic foams with multiscale structures are fabricated, demonstrating low adhesive superhydrophobic self-cleaning, striking loading capacity, stable corrosion resistance, and oil/water separation.

Development of a biaxial compression device for biological samples : preliminary experimental results for a closed cell foam

Biological tissues are subjected to complex loading states in vivo and in order to define constitutive equations that effectively simulate their mechanical behaviour under these loads, it is necessary to obtain data on the tissue's response to multiaxial loading. Single axis and shear testing of biological tissues is often carried out, but biaxial testing is less common. We sought to design and commission a biaxial compression testing device, capable of obtaining repeatable data for biological samples. The apparatus comprised a sealed stainless steel pressure vessel specifically designed such that a state of hydrostatic compression could be created on the test specimen while simultaneously unloading the sample along one axis with an equilibrating tensile pressure. Thus a state of equibiaxial compression was created perpendicular to the long axis of a rectangular sample. For the purpose of calibration and commissioning of the vessel, rectangular samples of closed cell ethylene vinyl acetate (EVA) foam were tested. Each sample was subjected to repeated loading, and nine separate biaxial experiments were carried out to a maximum pressure of 204 kPa (30 psi), with a relaxation time of two hours between them. Calibration testing demonstrated the force applied to the samples had a maximum error of 0.026 N (0.423% of maximum applied force). Under repeated loading, the foam sample demonstrated lower stiffness during the first load cycle. Following this cycle, an increased stiffness, repeatable response was observed with successive loading. While the experimental protocol was developed for EVA foam, preliminary results on this material suggest that this device may be capable of providing test data for biological tissue samples. The load response of the foam was characteristic of closed cell foams, with consolidation during the early loading cycles, then a repeatable load-displacement response upon repeated loading. The repeatability of the test results demonstrated the ability of the test device to provide reproducible test data and the low experimental error in the force demonstrated the reliability of the test data.

The role of HPC facilies in development of a novel road safety barrier

Water-filled portable road safety barriers are a common fixture in road works, however their use of water can be problematic, both in terms of the quantity of water used and the transportation of the water to the installation site. This project aims to develop a new design of portable road safety barrier, which will make novel use of composite and foam materials in order to reduce the barrier’s reliance on water in order to control errant vehicles. The project makes use of finite element (FE) techniques in order to simulate and evaluate design concepts. FE methods and models that have previously been tested and validated will be used in combination in order to provide the most accurate numerical simulations available to drive the project forward. LS-DYNA code is as highly dynamic, non-linear numerical solver which is commonly used in the automotive and road safety industries. Several complex materials and physical interactions are to be simulated throughout the course of the project including aluminium foams, composite laminates and water within the barrier during standardised impact tests. Techniques to be used include FE, smoothed particle hydrodynamics (SPH) and weighted multi-parameter optimisation techniques. A detailed optimisation of several design parameters with specific design goals will be performed with LS-DYNA and LS-OPT, which will require a large number of high accuracy simulations and advanced visualisation techniques. Supercomputing will play a central role in the project, enabling the numerous medium element count simulations necessary in order to determine the optimal design parameters of the barrier to be performed. Supercomputing will also allow the development of useful methods of visualisation results and the production of highly detailed simulations for end-product validation purposes. Efforts thus far have been towards integrating various numerical methods (including FEM, SPH and advanced materials models) together in an efficient and accurate manner. Various designs of joining mechanisms have been developed and are currently being developed into FE models and simulations.

Assimilating cell sheets and hybrid scaffolds for dermal tissue engineering

Cell sheets can be used to produce neo-tissue with mature extracellular matrix. However, extensive contraction of cell sheets remains a problem. We devised a technique to overcome this problem and applied it to tissue engineer a dermal construct. Human dermal fibroblasts were cultured with poly(lactic-co-glycolic acid)-collagen meshes and collagen-hyaluronic acid foams. Resulting cell sheets were folded over the scaffolds to form dermal constructs. Human keratinocytes were cultured on these dermal constructs to assess their ability to support bilayered skin regeneration. Dermal constructs produced with collagen-hyaluronic acid foams showed minimal contraction, while those with poly(lactic-co-glycolic acid)-collagen meshes curled up. Cell proliferation and metabolic activity profiles were characterized with PicoGreen and AlamarBlue assays, respectively. Fluorescent labeling showed high cell viability and F-actin expression within the constructs. Collagen deposition was detected by immunocytochemistry and electron microscopy. Transforming Growth Factor-alpha and beta1, Keratinocyte Growth Factor and Vascular Endothelial Growth Factor were produced at various stages of culture, measured by RT-PCR and ELISA. These results indicated that assimilating cell sheets with mechanically stable scaffolds could produce viable dermal-like constructs that do not contract. Repeated enzymatic treatment cycles for cell expansion is unnecessary, while the issue of poor cell seeding efficiency in scaffolds is eliminated.

A poly(D,L-lactide) resin for the preparation of tissue engineering scaffolds by stereolithography

Porous polylactide constructs were prepared by stereolithography, for the first time without the use of reactive diluents. Star-shaped poly(D,L-lactide) oligomers with 2, 3 and 6 arms were synthesised, end-functionalised with methacryloyl chloride and photocrosslinked in the presence of ethyl lactate as a non-reactive diluent. The molecular weights of the arms of the macromers were 0.2, 0.6, 1.1 and 5 kg/mol, allowing variation of the crosslink density of the resulting networks. Networks prepared from macromers of which the molecular weight per arm was 0.6 kg/mol or higher had good mechanical properties, similar to linear high molecular weight poly(D,L-lactide). A resin based on a 2-armed poly(D,L-lactide) macromer with a molecular weight of 0.6 kg/mol per arm (75 wt%), ethyl lactate (19 wt%), photo-initiator (6 wt%), inhibitor and dye was prepared. Using this resin, films and computer-designed porous constructs were accurately fabricated by stereolithography. Pre-osteoblasts showed good adherence to these photocrosslinked networks. The proliferation rate on these materials was comparable to that on high molecular weight poly(D,L-lactide) and tissue culture polystyrene.

Designed tissue engineering scaffolds prepared by stereolithography

For the fabrication of tissue engineering scaffolds, the intended tissue formation process imposes requirements on the architecture. The chosen porosity often is a tradeoff between volume and surface area accessible to cells, and mechanical properties of the construct. Interconnectivity of the pores is essential for cell migration through the scaffold and for mass transport. Conventional techniques such as salt leaching often result in heterogeneous structures and do not allow for a precise control of the architecture. Stereolithography is a rapid prototyping method that can be utilised to make 3D constructs with high spatial control by radical photopolymerisation. In this study, a regular structure based on cyclic repetition of cell units were designed through CAD modelling.. One of these structures was built on a stereolithography apparatus (SLA). Furthermore, a polylactide-based resin was developed that can be applied in stereolithography. Polylactide has proven before to be a well-performing polymer in bone tissue engineering. The final objective in this study is to build newly designed PDLLA scaffolds with a precise SLA fabrication technique to study the effect of scaffold architecture on mechanical and biological properties.

Poly(D,L-lactide)/hydroxyapatite composite tissue engineering scaffolds prepared by stereolithography

Hydroxyapatite (HAP) is a major component of bone and has osteoconductive and -inductive properties. It has been successfully applied as a substrate in bone tissue engineering, either with or without a biodegradable polymer such as polycaprolactone or polylactide. Recently, we have developed a stereolithography resin based on poly(D,L-lactide) (PDLLA) and a non-reactive diluent, that allows for the preparation of tissue engineering scaffolds with designed architectures. In this work, designed porous composite structures of PDLLA and HAP are prepared by stereolithography.

Testing and simulation of extruded polystyrene foam at low to moderate strain rates

This paper presents a study into the behaviour of extruded polystyrene foam at low strain rates. The foam is being studied in order assess its potential for use as part of a new innovative design of portable road safety barrier the aim to consume less water and reduce rates of serious injury. The foam was tested at a range of low strain rates, with the stress and strain behaviour of the foam specimens being recorded. The energy absorption capabilities of the foam were assessed as well as the response of the foam to multiple loadings. The experimental data was then used to create a material model of the foam for use in the explicit finite element solver LS-DYNA. Simulations were carried out using the material model which showed excellent correlation between the numerical material model and the experimental data.

Brominated flame retardants in the Australian population : 1993-2009

Brominated flame retardants, including hexabromocyclododecane (HBCD) and polybrominated diphenyl ethers (PBDEs) are used to reduce the flammability of a multitude of electrical and electronic products, textiles and foams. The use of selected PBDEs has ceased, however, use of decaBDE and HBCD continues. While elevated concentrations of PBDEs in humans have been observed in Australia, no data is available on other BFRs such as HBCD. This study aimed to provide background HBCD concentrations from a representative sample of the Australian population and to assess temporal trends of HBCD and compare with PBDE concentrations over a 16 year period. Samples of human milk collected in Australia from 1993 to 2009, primarily from primiparae mothers were combined into 12 pools from 1993 (2 pools); 2001; 2002/2003 (4 pools); 2003/2004; 2006; 2007/2008 (2 pools); and 2009. Concentrations of ∑HBCD ranged from not quantified (nq) to 19 ng g−1 lipid while α-HBCD and γ-HBCD ranged from nq to 10 ng g−1 lipid and nq to 9.2 ng g−1 lipid. β-HBCD was detected in only one sample at 3.6 ng g−1 lipid while ∑4PBDE ranged from 2.5 to 15.8 ng g−1 lipid. No temporal trend was apparent in HBCD concentrations in human milk collected in Australia from 1993 to 2009. In comparison, PBDE concentrations in human milk show a peak around 2002/03 (mean ∑4PBDEs = 9.6 ng g−1 lipid) and 2003/04 (12.4 ng g−1 lipid) followed by a decrease in 2007/08 (2.7 ng g−1 lipid) and 2009 (2.6 ng g−1 lipid). In human blood serum samples collected from the Australian population, PBDE concentrations did not vary greatly (p = 0.441) from 2002/03 to 2008/09. Continued monitoring including both human milk and serum for HBCD and PBDEs is required to observe trends in human body burden of HBCD and PBDEs body burden following changes to usage.

Dry powder inhaler formulations : effect of carrier size on the drug dispersion

Background: The size of the carrier influences drug aerosolization from a dry powder inhaler (DPI) formulation. Lactose particles with irregular shape and rough surface in a variety of sizes are additionally used as carriers; however, contradictory reports exist regarding the effect of carrier size on the dispersion of drug. We examined the influence of the spherical particle size of the biodegradable polylactide-co-glycolide (PLGA) carrier on the aerosolization of a model drug, salbutamol sulphate (SS). Methods: Four different sizes (20-150 µm) of polymer carriers were fabricated using solvent evaporation technique and the dispersion of SS from these carriers was measured by a Twin Stage Impinger (TSI). The size and morphological properties of polymer carriers were determined by laser diffraction and SEM, respectively. Results: The FPF was found to increase from 5.6% to 21.3% with increasing carrier sizeup to150 µm. Conclusions: The aerosolization of drug increased linearly with the size of polymer carriers. For a fixed mass of drug particles in a formulation, the mass of drug particles per unit area of carriers is higher in formulations containing the larger carriers, which leads to an increase in the dispersion of drug due to the increased mechanical forces occurred between the carriers and the device walls.

Fire safety of buildings based on realistic fire time-temperature curves

In recent times, fire has become a major disaster in buildings due to the increase in fire loads, as a result of modern furniture and light weight construction. This has caused problems for safe evacuation and rescue activities, and in some instances lead to the collapse of buildings (Lewis, 2008 and Nyman, 2002). Recent research has shown that the actual fire resistance of building elements exposed to building fires can be less than their specified fire resistance rating (Lennon and Moore, 2003, Jones, 2002, Nyman, 2002 and Abecassis-Empis et al. 2008). Conventionally the fire rating of building elements is determined using fire tests based on the standard fire time-temperature curve given in ISO 834. This ISO 834 curve was developed in the early 1900s, where wood was the basic fuel source. In reality, modern buildings make use of thermoplastic materials, synthetic foams and fabrics. These materials are high in calorific values and increase both the speed of fire growth and heat release rate, thus increasing the fire severity beyond that of the standard fire curve. Hence it suggests the need to use realistic fire time-temperature curves in tests. Real building fire temperature profiles depend on the fuel load representing the combustible building contents, ventilation openings and thermal properties of wall lining materials. Fuel load is selected based on a review and suitable realistic fire time-temperature curves were developed. Fire tests were then performed for plasterboard lined light gauge steel framed walls for the developed realistic fire curves. This paper presents the details of the development of suitable realistic building fire curves, and the fire tests using them. It describes the fire performance of tested walls in comparison to the standard fire tests and highlights the differences between them. This research has shown the need to use realistic fire exposures in assessing the fire resistance rating of building elements.

Experimental impact and finite element analysis of a composite, portable road safety barrier

This thesis provides an experimental and computational platform for investigating the performance and behaviour of water filled, plastic portable road safety barriers in an isolated impact scenario. A schedule of experimental impact tests were conducted assessing the impact response of an existing design of road safety barrier utilising a novel horizontal impact testing system. A coupled finite element and smooth particle hydrodynamic model of the barrier system was developed and validated against the results of the experimental tests. The validated model was subsequently used to assess the effect of certain composite materials on the impact performance of the water filled, portable road safety barrier system.

Numerical simulations of particle deposition in metal foam heat exchangers

Australia is a high-potential country for geothermal power with reserves currently estimated in the tens of millions of petajoules, enough to power the nation for at least 1000 years at current usage. However, these resources are mainly located in isolated arid regions where water is scarce. Therefore, wet cooling systems for geothermal plants in Australia are the least attractive solution and thus air-cooled heat exchangers are preferred. In order to increase the efficiency of such heat exchangers, metal foams have been used. One issue raised by this solution is the fouling caused by dust deposition. In this case, the heat transfer characteristics of the metal foam heat exchanger can dramatically deteriorate. Exploring the particle deposition property in the metal foam exchanger becomes crucial. This paper is a numerical investigation aimed to address this issue. Two dimensional (2D) numerical simulations of a standard one-row tube bundle wrapped with metal foam in cross-flow are performed and highlight preferential particle deposition areas.