Development of a novel 3D culture system for screening features of a complex implantable device for CNS repair

Tubular scaffolds which incorporate a variety of micro- and nanotopographies have a wide application potential in tissue engineering especially for the repair of spinal cord injury (SCI). We aim to produce metabolically active differentiated tissues within such tubes, as it is crucially important to evaluate the biological performance of the three-dimensional (3D) scaffold and optimize the bioprocesses for tissue culture. Because of the complex 3D configuration and the presence of various topographies, it is rarely possible to observe and analyze cells within such scaffolds in situ. Thus, we aim to develop scaled down mini-chambers as simplified in vitro simulation systems, to bridge the gap between two-dimensional (2D) cell cultures on structured substrates and three-dimensional (3D) tissue culture. The mini-chambers were manipulated to systematically simulate and evaluate the influences of gravity, topography, fluid flow, and scaffold dimension on three exemplary cell models that play a role in CNS repair (i.e., cortical astrocytes, fibroblasts, and myelinating cultures) within a tubular scaffold created by rolling up a microstructured membrane. Since we use CNS myelinating cultures, we can confirm that the scaffold does not affect neural cell differentiation. It was found that heterogeneous cell distribution within the tubular constructs was caused by a combination of gravity, fluid flow, topography, and scaffold configuration, while cell survival was influenced by scaffold length, porosity, and thickness. This research demonstrates that the mini-chambers represent a viable, novel, scale down approach for the evaluation of complex 3D scaffolds as well as providing a microbioprocessing strategy for tissue engineering and the potential repair of SCI

Equivalence of using a desktop virtual reality science simulation at home and in class

The use of virtual laboratories is growing as companies and educational institutions try to expand their reach, cut costs, increase student understanding, and provide more accessible hands on training for future scientists. Many new higher education initiatives outsource lab activities so students now perform them online in a virtual environment rather than in a classroom setting, thereby saving time and money while increasing accessibility. In this paper we explored whether the learning and motivational outcomes of interacting with a desktop virtual reality (VR) science lab simulation on the internet at home are equivalent to interacting with the same simulation in class with teacher supervision. A sample of 112 (76 female) university biology students participated in a between-subjects experimental design, in which participants learned at home or in class from the same virtual laboratory simulation on the topic of microbiology. The home and classroom groups did not differ significantly on post-test learning outcome scores, or on self-report measures of intrinsic motivation or self-efficacy. Furthermore, these conclusions remained after accounting for prior knowledge or goal orientation. In conclusion, the results indicate that virtual simulations are learning activities that students can engage in just as effectively outside of the classroom environment

Friction of flat and micropatterned interfaces with nanoscale roughness

The dry friction of surfaces with nanoscale roughness and the possibility of using micropatterning to tailor friction by manipulating contact area is investigated. Square wave patterns produced on samples from silicon wafers (and their unstructured equivalent) were slid against unstructured silicon counter surfaces. The width of the square wave features was adjusted to vary the apparent feature contact area. The existence of nanoscale roughness was sufficient to ensure Amontons’ first law (F = μP) on both structured & unstructured samples. Somewhat counterintuitively, friction was independent of the apparent feature contact area making it difficult to tailor friction via the feature contact area. This occurred because, even though the apparent feature contact area was adjusted, the surface roughness and nominal flatness at the contact interface was preserved ensuring that the real contact area and thereby the friction, were likewise preserved. This is an interesting special case, but not universally applicable: friction can indeed be adjusted by structuring provided the intervention leads to a change in real contact area (or interlocking)– and this depends on the specific surface geometry and topography

Protein Expression of STRO-1 Cells in Response to Different Topographic Features

Human skeletal stem cells (STRO-1 positive) display distinct responses to different topographical features. On a flat surface, skeletal cells spread, and in vitro, they typically display a polarized, fibroblast-like morphology. However, on microgrooved surfaces, these cells prefer to stretch along the grooves forming a similar morphology to in vivo, bipolarized fibroblasts. In contrast, on nanopits, these cells display a polygonal and osteoblastic phenotype. We have examined mechanotransduction events of STRO-1 positive in response to fibroblastic, microgrooved and osteogenic, controlled disorder nanopit, topographies using proteomics after 3 days in culture. Protein expression profiles were analyzed by difference gel electrophoresis to identify proteins that showed modulation of expression in response to different topographic features to assess early decision events in these cells on these discrete topographies. After only 72 hours in culture, STRO-1 positive displayed differential regulations of families of proteins involved in cell migration and proliferation. The current study indicated that osteogenic decision specific events had already occurred. Runx2 was localized in nuclei of the skeletal stem cells on the osteogenic nanopits; however, few signaling pathway changes were observed. This study demonstrated that micro- and nanotopographies activated skeletal stem cells at different times and with distinct mechanotransduction profiles

Nacre Topography Produces Higher Crystallinity in Bone than Chemically Induced Osteogenesis

It is counter-intuitive that invertebrate shells can induce bone formation yet nacre, or mother of pearl, from marine shells is both osteoinductive and osteointegrative. Nacre is composed of aragonite (calcium carbonate) and induces production of vertebrate bone (calcium phosphate). Exploited by the Mayans for dental implants, this remarkable phenomenon has been confirmed in vitro and in vivo yet the characteristic of nacre that induces bone formation remains unknown. By isolating nacre topography from its inherent chemistry in the production of polycaprolactone (PCL) nacre replica, we show that, for mesenchymal stem cells, nacre topography is osteoinductive. Gene expression of specific bone marker proteins, osteopontin, osteocalcin, osteonectin and osterix are increased 10-, 2- 1.7- and 1.8-fold respectively when compared to planar PCL. Furthermore, we demonstrate that bone tissue that forms in response to the physical topographical features of nacre has higher crystallinity than bone formed in response to chemical cues with full width half maximum for PO4 3- Raman shift of 7.6±0.7 for mineral produced in response to nacre replica compared to a much broader 34.6±10.1 in response to standard osteoinductive medium. These differences in mineral product are underpinned by differences in cellular metabolism. This observation can be exploited in the design of bone therapies; a matter that is most pressing in light of a rapidly ageing human population. Aragonite and calcite are the two calcium carbonate polymorphs that constitute the shell of molluscan bivalves conferring strength and resilience due to the nano- and microstructural assembly of the overall architecture. A small percentage of the invertebrate shell constitute the organic matrix which is responsible for the intricate processes of nucleation, growth and inhibition of calcium carbonate crystals resulting in the well-defined shell structure. The discovery of fully integrated shell dental implants in Mayan skulls initiated a number of studies showing that nacre, or mother of pearl, the aragonite calcium carbonate polymorph derived from the pearl oyster Pinctada maxima has good osteointegrative properties in vivo. Further exploration of this phenomenon in human jaw reconstructions and sheep femur implants confirm the osteointegrative properties of invertebrate shells. In addition, nacre initiates osteogenic differentiation in mesenchymal stem cells (MSCs) in vitro. This observation has led to a number of studies in which nacre and its chemistry have been incorporated into the design of existing biomaterials to induce bone formation. MSCs can be induced into undergoing osteogenesis in vitro by the use of pre-formulated soluble factors in the culture media, chemically defined surfaces, substrate matrix elasticity and the surface topography of the substrate. These approaches induce osteogenesis when presented in isolation or in combination. When these cues are presented in combination, surface patterning plays an important role and topography can have a stronger influence on cell behaviour when presented with effective surface chemistries. In vertebrate and invertebrate systems, the main requisites for forming hard tissue or biomineral structures are calcium phosphate and calcium carbonate respectively, both of which are assembled in a variety of ways generating an incredible amount of structural diversity. This juxtaposition of phosphate and carbonate is described as the “Bone-Shell Divide”. It is intriguing that mammalian cells respond to mineral on the shell side of the Bone-Shell Divide and this begs questions: which feature of nacre elicits this response and, in transcending the Bone-Shell Divide, do MSCs produce bone of similar or superior characteristics to that induced by other means? Addressing these questions has important implications in tissue engineering and biomaterial applications, especially with regards to orthopaedic applications where critical sized defects in trauma and reconstructive surgery demand large areas of intact bone usually acquired by creating a secondary injury site. By isolating the topographical features of nacre from its inherent chemistry, we show that the osteoinductive properties of nacre arise from the patterning of the surface presented to MSCs. Importantly, separating nacre topography from its inherent chemistry enhances the osteogenic response. In this report we dissect out the contribution of topography to nacre bioactivit

The static friction peak in reciprocating sliding

This paper investigates why the static friction peak is mostly absent in reciprocating sliding and gross-slip fretting literature. Here, reciprocating sliding tests were conducted on ultra-smooth silicon surfaces. A prominent static friction peak was present in the initial cycles. However, a rapid wear-induced decay in the static friction peak occurred after the first cycle with the peak being mostly absent by about 30 cycles. Two possible explanations are proposed for the wear-induced decay: (1) that increasing surface roughness (with cycles) reduces the fully stuck contact area and (2) that wear reduces the bonding strength of the stuck interface by removing third body contaminant molecules. Predictions from a multi-asperity friction model are used to support these arguments

Nanotopographical induction of osteogenesis through adhesion, bone morphogenic protein cosignaling, and regulation of microRNAs

It is emerging that nanotopographical information can be used to induce osteogenesis from mesenchymal stromal cells from the bone marrow and it is hoped that this nanoscale bioactivity can be utilized to engineer next generation implants. However, the osteogenic mechanism of surfaces is currently poorly understood. In this report, we investigate mechanism and implicate bone morphogenic protein (BMP) in up-regulation of RUNX2 and show that RUNX2 and its regulatory miRNAs are BMP sensitive. Our data demonstrates that osteogenic nanotopography promotes co-localization of intergrins and BMP2 receptors in order to enhance osteogenic activity and that vitronectin is important in this interface. This provides insight that topographical regulation of adhesion can have effects on signaling cascades outside of cytoskeletal signaling and that adhesions can have roles in augmenting BMP signaling

Characterization techniques for studying the properties of nanocarriers for systemic delivery

Nanocarriers have attracted a huge interest in the last decade as efficient drug delivery systems and diagnostic tools. They enable effective, targeted, controlled delivery of therapeutic molecules while lowering the side effects caused during the treatment. The physicochemical properties of nanoparticles determine their in vivo pharmacokinetics, biodistribution and tolerability. The most analyzed among these physicochemical properties are shape, size, surface charge and porosity and several techniques have been used to characterize these specific properties. These different techniques assess the particles under varying conditions, such as physical state, solvents etc. and as such probe, in addition to the particles themselves, artifacts due to sample preparation or environment during measurement. Here, we discuss the different methods to precisely evaluate these properties, including their advantages or disadvantages. In several cases, there are physical properties that can be evaluated by more than one technique. Different strengths and limitations of each technique complicate the choice of the most suitable method, while often a combinatorial characterization approach is needed

Global patient outcomes after elective surgery: prospective cohort study in 27 low-, middle- and high-income countries.

BACKGROUND: As global initiatives increase patient access to surgical treatments, there remains a need to understand the adverse effects of surgery and define appropriate levels of perioperative care. METHODS: We designed a prospective international 7-day cohort study of outcomes following elective adult inpatient surgery in 27 countries. The primary outcome was in-hospital complications. Secondary outcomes were death following a complication (failure to rescue) and death in hospital. Process measures were admission to critical care immediately after surgery or to treat a complication and duration of hospital stay. A single definition of critical care was used for all countries. RESULTS: A total of 474 hospitals in 19 high-, 7 middle- and 1 low-income country were included in the primary analysis. Data included 44 814 patients with a median hospital stay of 4 (range 2-7) days. A total of 7508 patients (16.8%) developed one or more postoperative complication and 207 died (0.5%). The overall mortality among patients who developed complications was 2.8%. Mortality following complications ranged from 2.4% for pulmonary embolism to 43.9% for cardiac arrest. A total of 4360 (9.7%) patients were admitted to a critical care unit as routine immediately after surgery, of whom 2198 (50.4%) developed a complication, with 105 (2.4%) deaths. A total of 1233 patients (16.4%) were admitted to a critical care unit to treat complications, with 119 (9.7%) deaths. Despite lower baseline risk, outcomes were similar in low- and middle-income compared with high-income countries. CONCLUSIONS: Poor patient outcomes are common after inpatient surgery. Global initiatives to increase access to surgical treatments should also address the need for safe perioperative care. STUDY REGISTRATION: ISRCTN5181700