Australia's micro/nanofabrication research environment has been significantly enhanced over the past decade by the federally initiated provision of open-access research infrastructure, through the National Collaborative Research Infrastructure Strategy (NCRIS). NCRIS has become a cornerstone of the research sector, enhancing innovation and industry engagement, and benefits the Australian economy in broader terms. The Strategy oversees 27 platforms that operate independently, each providing a different type of world-class research infrastructure accessible by scientists and engineers. The services range from supercomputer time, to procurement of marine observational datasets, to access of advanced imaging and analysis processes. One of the first NCRIS platforms was the Australian National Fabrication Facility (ANFF), which gives access to micro/nanofabrication equipment. Access to ANFF enables clients to manipulate advanced materials, transforming them into micro/nanoscale structures with applications in energy harvesting, pharmaceutical and medical devices, sensors, and nanoelectronics. ANFF is supported by long-term grants for operating, and capital expenses from NCRIS, supplemented by co-investment from state governments, universities, and government labs such as the Commonwealth Scientific & Industrial Research Organisation (CSIRO). This financial support cumulatively amounts to more than US$400M, keeping micro/nanofabrication infrastructure at the cutting edge, and ensuring that world-class expertise is maintained. ANFF's portfolio includes some 500 micro/nanofabrication capabilities, and a pool of over 100 process and tool experts. ANFF engineers servicing national and overseas customers can perform contract work for industry clients, give advanced equipment training and on-tool assistance and they maintain peak performance of equipment. The Network was founded in 2007 and has since grown to include hubs at 21 Australian universities– hubs that are thematically grouped into eight ANFF nodes. This distributed model harnesses the critical mass of each host university's research environment while reducing the need to duplicate infrastructure. Website: https://www.anff.org.au/. Melbourne Centre for Nanofabrication (MCN) is largest of the seven Victoria-based hubs that form the ANFF-Vic Node. The MCN is an ISO9001-certified facility, with the means to produce complex micro- and nanoscience-based demonstration devices for universities and industry using an array of tools, and provide expertise on, and tools for, high-throughput validation, and upscaling of various systems including etching, lithography, and thin-film coatings. Recently the MCN has demonstrated a new nanofabrication route to manufacture arrays of optically-transparent polymeric nanoneedles facilitating delivery of genes to immune cells.[1] The node has also produced a novel approach to imprint micro- and nanoscale topographical features into conventional cell cultureware, enabling high-throughput screening to interrogate the effects of surface topographies into unique cell specific responses and fate determination.[2] The MCN supports the development of fully solution-processed transparent inorganic QLEDs,[3] and an ultrathin material—tin mono-sulfide nanosheets—for application in energy conversion.[4] Website: https://nanomelbourne.com/. The ANFF-ACT Node at the Australian National University (ANU) is at the forefront of the next generation optoelectronic devices, nano-photonics and metamaterial-based optical devices. The Node forms an integrated capability for III-V semiconductor optoelectronic devices, from epitaxial growth, device design and fabrication to characterisation. The Node is also co-located with the headquarters of the Australian Research Council Centre of Excellence for Transformative Meta-Optical Systems, which aims to overcome complex challenges in light generation, manipulation and detection at the nanoscale, thereby aligning optics with the advances in nanoelectronics. Recent achievements include (i) the design of a hybrid chalcogenide-germanosilicate waveguide platform for highly efficient, robust, and reflection-free light coupling;[5] and (ii) semiconductor nanowire arrays for NO2 sensing operating at room temperature down to the ppb level with outstanding selectivity and long-term stability.[6] Website: http://anff-act.anu.edu.au/. The ANFF Materials Node at the Universities of Wollongong and Newcastle fabricates both hard and soft materials. The two sites are highly specialised: Wollongong established platforms for 3D bioprinted skin constructs, especially 3D printed cell-laden platforms for skin regeneration.[7] Newcastle is involved in late-stage development of a solar cell technology that can be created in abundance using roll-to-roll printers, and fitted to any surface with double-sided sticky-tape. Website: https://www.anffmaterials.org/. The ANFF-NSW node has hubs at UNSW and the University of Sydney that offer a range of clean-rooms supporting advanced electron-beam lithography and silicon-MOS process lines enabling fabrication of optical chips, electronic devices, and quantum technologies. A key strength of the NSW Node has been its support of the development of silicon-based qubits for quantum computing.[8] Ultra-shallow all-epitaxial aluminium gate GaAs/AlxGa1−xAs transistors were with high electron mobility[9] using the Node's nanofabrication capabilities. Website: https://www.anff-nsw.org/. The ANFF Optofab Node is the most distributed facility with locations at Macquarie University, the Universities of Adelaide and Sydney, the University of Technology Sydney, and the ANU. This node has expertise in laser-machined precision optics, and can produce and process fibres, and planar and bulk materials. The node has led the work of developing an unprecedented approach of novel designer glasses that is projected to serve as an optical platform to build the next generation devices aiming for high density integration and realization of novel concepts.[10] The target application of such glasses include, but are not limited to, LIDAR for autonomous vehicles, mid-infrared integrated devices for life and environmental science applications and next generation integrated quantum photonic devices. http://optofab.org.au/. The ANFF-Q Node at the University of Queensland and Griffith University are experts at the interface of biology and the fabricated devices, including microfluidics and the science behind organic solar cells. For example, the team has been heavily involved in developing a novel nanomechanical resonator for a wide range of technological applications [11] https://anff-q.org.au/. The ANFF-SA Node at the University of South Australia and Flinders University specialises in the design and prototyping of lab-on-a-chip technologies and medical devices. Research highlights include the development of a silk-based optical fibre coating for the production of bifunctional pH sensing and nanoscale optical-coherence tomography (OCT) imaging probes, and their application for in vitro fertilisation (IVF) revealing the presence of oocytes within ovarian follicles.[12] https://www.anff-sa.com/. The ANFF-WA Node at the University of Western Australia is a vertically integrated facility that focuses on microelectromechanical systems and infrared technologies. The node has enabled the development of a low size, weight and power solution for delivering mechanically robust spectrally selective sensing suitable for deployment on field-portable platforms that are in increasing demand.[13] https://www.mrg.uwa.edu.au/. In summary, ANFF's eight nodes serve a consistently growing client base whose interests cover the full range of national research priorities and the applications that research can have in economic recovery. The projects enabled by ANFF have matured to the point where the fabricated part is only a small portion of the incredible research outcomes reported.

Current clinical delivery of recombinant human bone morphogenetic proteins (rhBMPs) utilises freeze-dried collagen. Despite effective new bone generation, rhBMP via collagen can be limited by significant complications due to inflammation and uncontrolled bone formation. This study aimed to produce an alternative rhBMP local delivery system to permit more controllable and superior rhBMP-induced bone formation. Cylindrical porous poly(lactic-co-glycolic acid) (PLGA) scaffolds were manufactured by thermally-induced phase separation. Scaffolds were encapsulated with anabolic rhBMP-2 (20 µg) ± anti-resorptive agents: zoledronic acid (5 µg ZA), ZA pre-adsorbed onto hydroxyapatite microparticles, (5 µg ZA/2% HA) or IkappaB kinase (IKK) inhibitor (10 µg PS-1145). Scaffolds were inserted in a 6-mm critical-sized femoral defect in Wistar rats, and compared against rhBMP-2 via collagen. The regenerate region was examined at 6 weeks by 3D microCT and descriptive histology. MicroCT and histology revealed rhBMP-induced bone was more restricted in the PLGA scaffolds than collagen scaffolds (-92.3% TV, p < 0.01). The regenerate formed by PLGA + rhBMP-2/ZA/HA showed comparable bone volume to rhBMP-2 via collagen, and bone mineral density was +9.1% higher (p < 0.01). Local adjunct ZA/HA or PS-1145 significantly enhanced PLGA + rhBMP-induced bone formation by +78.2% and +52.0%, respectively (p ≤ 0.01). Mechanistically, MG-63 human osteoblast-like cells showed cellular invasion and proliferation within PLGA scaffolds. In conclusion, PLGA scaffolds enabled superior spatial control of rhBMP-induced bone formation over clinically-used collagen. The PLGA scaffold has the potential to avoid uncontrollable bone formation-related safety issues and to customise bone shape by scaffold design. Moreover, local treatment with anti-resorptive agents incorporated within the scaffold further augmented rhBMP-induced bone formation.

We describe two studies encompassing the iterative refinement of a polymer-based rhBMP-2 delivery system for bone tissue engineering. Firstly, we compared the bone-forming capacity of porous poly(D,L-lactic-co-glycolic acid) (PLGA) scaffolds produced by thermally induced phase separation (TIPS) with non-porous solvent cast poly(D,L-lactic acid) (PDLLA) used previously. Secondly, we examined the potential synergy between rhBMP-2 and local bisphosphonate in the PLGA scaffold system.In vivo ectopic bone formation studies were performed in C57BL6/J mice. Polymer scaffolds containing 0, 5, 10 or 20 µg rhBMP-2 were inserted into the dorsal musculature. At all rhBMP-2 doses, porous PLGA produced significantly higher bone volume (BV, mm3) than the solid PDLLA scaffolds. Next, porous PLGA scaffolds containing 10 µg rhBMP-2 ± 0.2, or 2 µg zoledronic acid (ZA) were inserted into the hind-limb musculature. Co-delivery of local 10 µg rhBMP-2/2 µg ZA significantly augmented bone formation compared with rhBMP-2 alone (400 % BV increase, p < 0.01). Hydroxyapatite microparticle (HAp) addition (2 % w/w) to the 10 µg rhBMP-2/0.2 µg ZA group increased BV (200 %, p < 0.01). We propose that this was due to controlled ZA release of HAp-bound ZA. Consistent with this, elution analyses showed that HAp addition did not alter the rhBMP-2 elution, but delayed ZA release. Moreover, 2 % w/w HAp addition reduced the scaffold’s compressive properties, but did not alter ease of surgical handling.In summary, our data show that refinement of the polymer selection and scaffold fabrication can enhance rhBMP-2 induced bone formation in our bone tissue engineering implant, and this can be further optimised by the local co-delivery of ZA/HAp.

This paper describes the use of a microfluidic device to produce porous, hierarchical nano-patterned microspheres. Within this device, explicit control over the micro-phase separation and macro-phase solidification behaviour of a polystyrene-polyethylene oxide di-block copolymer dissolved in an organic (polar) solvent droplet has been achieved through the tailoring of the staged onset of diffusion induced phase separation (DIPS) and thermally induced phase separation (TIPS) events post dispersion in an aqueous continuous phase. Due to water and the organic solvent (dimethylene carbonate (DMC)) being partially miscible, the organic solvent within the dispersed phase diffuses very slowly out of the polymer-loaded droplets into the continuous transport fluid (water) as they flow down the microchannel. This permits controlled shrinkage of the droplets (and hence sizing), and the eventual condensation of the external surface of the polymer microbeads, prior to the timed onset of TIPS, which promotes the formation of a highly porous internal structure (via liquid-liquid macrophase separation) within the microbeads. The thickness of the external layer can be controlled by varying the timeframe for DIPS, along with porosity of the internals of these hierarchical microbeads by varying the TIPS parameters. The resulting monodisperse hierarchical microspheres are comprised of external surfaces decorated with nano-scale self-assembled block copolymer domains, while the internal structure is highly porous and all surfaces display the same nano-scale domains. The hydrophilic nature of the PEO nano-domains results in these microparticles being near neutrally buoyant in water and bioactive moieties can be presented on the terminal ends of the PEO chains in each domain. These novel microparticles are thus seen as ideal for bead-based cell culture applications and the use of this methodology with degradable block copolymers ideal for future application in regenerative medicine.

Human embryonic stem cells (hESC) are expected to provide revolutionary therapeutic applications and drug discovery technologies. In order for this to be achieved a reproducible, defined animal component free culture system is required for the scale-up production of undifferentiated hESC. In this work we have investigated the applicability of a recombinantly produced domain of human vitronectin as an extracellular matrix alternative to the common standards Geltrex or Matrigel. In addition we have validated an ascorbate free media capable of supporting CD30(low) populations of hESC through a multi-factorial analysis of bFGF and Activin A. The recombinant vitronectin domain combined with the ascorbate free media were capable of supporting 3 cell lines, MEL1, MEL2 and hES3 for 10 or more passages while maintaining hESC pluripotency markers and differentiation capacity. The culture method outlined here provides a platform for future investigation into growth factor and extracellular matrix effects on hESC maintenance prior to bioreactor scale-up.

Unlocking the clinical potential of stem cell based therapies requires firstly elucidation of the biological mechanisms which direct stem cell fate decisions and thereafter, technical advances which allow these processes to be driven in a fully defined culture environment. Strategies for the generation of defined surfaces for human embryonic stem cell (hESC) and mesenchymal stem cell (MSC) culture remain in their infancy. In this paper we outline a simple, effective and efficient method for presenting proteins or peptides on an otherwise non-fouling Layer-by-Layer (LbL) self-assembled surface of hyaluronic acid (HA) and chitosan (CHI). We are able to generate a surface that has both good temporal stability and the ability to direct biological outcomes based on its defined surface composition. Surface functionalization is achieved through suspending the selected extracellular matrix (ECM) protein domain or extracted full-length protein in buffer containing a cross-linking agent (N-hydroxysulfosuccinimide/N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride) over the LbL HA-CHI surface and then allowing the solvent to evaporate overnight. This simple, but important step results in remarkable protein deposition efficiencies often exceeding 50%, whereas traditional cross-linking methods result in such poor deposition of non-collagenous proteins that a.) quantification of bound amounts of protein is outside the resolution of commonly utilized protein assays, and b.) these surfaces are both unable to support cell attachment and growth. The utility of the protein-modified HA-CHI surfaces is demonstrated through the identification of specific hESC attachment efficiencies and through directing MSC osteogenic outcomes on these fully defined surfaces. This simple and scalable method is shown to enable the development of defined stem cell culture conditions, as well as the elucidation of the fundamental biological processes necessary for the realization of stem cell based therapies.

The Dearing Report’s (1997) radical proposals challenged lecturers in higher education to develop innovative assessment strategies. This paper explores the dilemmas experienced by one teaching team in designing and implementing a student self‐assessment strategy within a community nursing degree programme. The paper reviews the impact on students’ sense of autonomy and critical thinking skills. In addition, it considers, in depth, the risks associated with developing and implementing a strategy involving self‐assessment. Drawing on a range of sources it examines the drivers for the initiative, the response from the range of stakeholders involved and the impact on the student experience. The academic team found that developing such a creative initiative is time consuming, provokes anxiety and requires extensive negotiation and collaboration between academic and practice colleagues. However, the adoption of a self‐assessment initiative has a significant effect on students’ critical thinking skills and warrants the effort.

Self-assessment is advocated as a means of encouraging learners to develop critical thinking skills. This article examines how self-assessment has been used within a community nursing programme to provoke learners’ integration of personal and professional development and thus extend their critical perspectives on their developing practice. The article explores how self-assessment, combined with a culture of reflective dialogue within learner-led discussion groups, enhances learners’ emotional engagement with learning. It suggests that self-assessment, used as the focus of interactive learning experiences, can provoke integrative learning within a professional degree programme. By legitimately bringing personal and professional knowledge and skills acquisition to the fore, learners engage in challenging reflective dialogue with tutors, peers and mentors in practice. The article argues that this provides a pivot for transformative learning. This report draws on the evaluation of the self-assessment strategy within a community nursing programme to illuminate the discussion. The evaluation draws on Joyes’ model of evaluation to include a review of both the process of, and stakeholder perspectives on, the development and implementation of the initiative. The community nursing case study suggests that self-assessment can help harness emotion to benefit learning.

A rapid antibody-based detection system has been developed for the presence of free trenbolone in bovine samples. Polyclonal antibodies were produced that showed specificity toward epitopes located around the steroidal A-ring of the trenbolone molecule. These antibodies were shown to have little or no recognition for many closely related compounds. The antibodies were utilized as the specific biorecognition molecules in competitive and inhibitive enzyme-linked immunosorbent assay systems. While both assays were able to detect low nanogram concentrations of trenbolone in bovine bile, the competitive format was more sensitive (2.41 vs 17.15 ng/mL for TRAb2 and 3.31 vs 30.73 ng/mL for TRAb1). This format was also more accurate and the data produced by this assay fitted more closely to the four parameter equation used to calculate the standard curve. This was a common finding with both of the polyclonal antibodies, suggesting that this was a characteristic of the format used.

Polyclonal and monoclonal antibody molecules have had a significant impact in analytical detection systems throughout the last few decades. Recently, the emergence of recombinant antibody technology is also playing a part in diagnostic applications. Recombinant antibody display technology is being exploited to produce antigen binding fragment's (Fab) of an antibody and single chain Fv (scFv) antibodies. These recombinant molecules can be as sensitive and specific as their parent molecules, and can often be produced relatively easy and cheaply. This manuscript will focus on antibody phage display technology, and other novel expression systems for the production of recombinant antibodies. It will also critically examine their applications in different assay formats, residue analysis, and, more recently, their role in proteomics.

There are many compounds that require analysis ranging from pesticide levels in corn to disease markers in human patients. There are copious challenges to be met when measuring analytes such as the matrix in which they are to be determined, the amounts present, the cost and the rapidity of the result required. Enzyme immunoassays, immunoaffinity chromatography, immunomagnetic polymerase chain reaction, flow cytometry and immunobiological biosensors have all characteristics that can enhance analytical techniques. Antibody-based methods have found applications in a large number of diverse areas such as food and water analysis, clinical diagnosis and therapeutics The structure and modes of production of antibodies and antibody-based derivatives is described and their applications in analysis critically examined.