Transplantation of human fetal blood stem cells in the osteogenesis imperfecta mouse leads to improvement in multiscale tissue properties.

Osteogenesis imperfecta (OI or brittle bone disease) is a disorder of connective tissues caused by mutations in the collagen genes. We previously showed that intrauterine transplantation of human blood fetal stem/stromal cells in OI mice (oim) resulted in a significant reduction of bone fracture. This work examines the cellular mechanisms and mechanical bone modifications underlying these therapeutic effects, particularly examining the direct effects of donor collagen expression on bone material properties. In this study, we found an 84% reduction in femoral fractures in transplanted oim mice. Fetal blood stem/stromal cells engrafted in bones, differentiated into mature osteoblasts, expressed osteocalcin, and produced COL1a2 protein, which is absent in oim mice. The presence of normal collagen decreased hydroxyproline content in bones, altered the apatite crystal structure, increased the bone matrix stiffness, and reduced bone brittleness. In conclusion, expression of normal collagen from mature osteoblast of donor origin significantly decreased bone brittleness by improving the mechanical integrity of the bone at the molecular, tissue, and whole bone levels.

The effect of an upstream turbine on a low-aspect ratio vane

The interaction between a high-pressure rotor and a downstream vane is dominated by vortex-blade interaction. Each rotor blade passing period two co-rotating vortex pairs, the tip-leakage and upper passage vortex and the lower passage and trailing shed vortex, impinge on, and are cut by, the vane leading edge. In addition to the streamwise vortex the tip-leakage flow also contains a large velocity deficit. This causes the interaction of the tip-leakage flow with a downstream vane to differ from typical vortex blade interaction. This paper investigates the effect these interaction mechanisms have on a downstream vane. The test geometry considered was a low aspect ratio second stage vane located within a S-shaped diffuser with large radius change mounted downstream of a shroudless high-pressure turbine stage. Experimental measurements were conducted at engine-representative Mach and Reynolds numbers, and data was acquired using a fast-response aerodynamic probe upstream and downstream of the vane. Time-resolved numerical simulations were undertaken with and without a rotor tip gap in order to investigate the relative magnitude of the interaction mechanisms. The presence of the upstream stage is shown to significantly change the structure of the secondary flow in the vane and to cause a small drop in its performance.

Unsteady ejectors: The effect of driver jet mark-space ratio

mark Unsteady ejectors can be driven by a wide range of driver jets. These vary from pulse detonation engines, which typically have a long gap between each slug of fluid exiting the detonation tube (mark-space ratios in the range 0.1-0.2) to the exit of a pulsejet where the mean mass flow rate leads to a much shorter gap between slugs (mark-space ratios in the range 2-3). The aim of this paper is to investigate the effect of mark-space ratio on the thrust augmentation of an unsteady ejector. Experimental testing was undertaken using a driver jet with a sinusoidal exit velocity profile. The mean value, amplitude and frequency of the velocity profile could be changed allowing the length to diameter ratio of the fluid slugs L/D and the mark-space ratio (the ratio of slug length to the spacing between slugs) L/S to be varied. The setup allowed L/S of the jet to vary from 0.8 to 2.3, while the L/D ratio of the slugs could take any values between 3.5 and 7.5. This paper shows that as the mark-space ratio of the driver jet is increased the thrust augmentation drops. Across the range of mark-space ratios tested, there is shown to be a drop in thrust augmentation of 0.1. The physical cause of this reduction in thrust augmentation is shown to be a decrease in the percentage time over which the ejector entrains ambient fluid. This is the direct result ofthe space between consecutive slugs in the driver jet decreasing. The one dimensional model reported in Heffer et al. [1] is extended to include the effect of varying L/S and is shown to accurately capture the experimentally measured behavior ofthe ejector. Copyright © 2010 by the American Institute of Aeronautics and Astronautics, Inc.

The application of STEM and in situ controlled dehydration to bacterial systems using ESEM.

Transmission imaging with an environmental scanning electron microscope (ESEM) (Wet STEM) is a recent development in the field of electron microscopy, combining the simple preparation inherent to ESEM work with an alternate form of contrast available through a STEM detector. Because the technique is relatively new, there is little information available on how best to apply this technique and which samples it is best suited for. This work is a description of the sample preparation and microscopy employed by the authors for imaging bacteria with Wet STEM (scanning transmission electron microscopy). Three different bacterial samples will be presented in this study: first, used as a model system, is Escherichia coli for which the contrast mechanisms of STEM are demonstrated along with the visual effects of a dehydration-induced collapse. This collapse, although clearly in some sense artifactual, is thought to lead to structurally meaningful morphological information. Second, Wet STEM is applied to two distinct bacterial systems to demonstrate the novel types of information accessible by this approach: the plastic-producing Cupriavidus necator along with wild-type and ΔmreC knockout mutants of Salmonella enterica serovar Typhimurium. Cupriavidus necator is shown to exhibit clear internal differences between bacteria with and without plastic granules, while the ΔmreC mutant of S. Typhimurium has an internal morphology distinct from that of the wild type.

Extracellular-matrix tethering regulates stem-cell fate.

To investigate how substrate properties influence stem-cell fate, we cultured single human epidermal stem cells on polydimethylsiloxane (PDMS) and polyacrylamide (PAAm) hydrogel surfaces, 0.1 kPa-2.3 MPa in stiffness, with a covalently attached collagen coating. Cell spreading and differentiation were unaffected by polydimethylsiloxane stiffness. However, cells on polyacrylamide of low elastic modulus (0.5 kPa) could not form stable focal adhesions and differentiated as a result of decreased activation of the extracellular-signal-related kinase (ERK)/mitogen-activated protein kinase (MAPK) signalling pathway. The differentiation of human mesenchymal stem cells was also unaffected by PDMS stiffness but regulated by the elastic modulus of PAAm. Dextran penetration measurements indicated that polyacrylamide substrates of low elastic modulus were more porous than stiff substrates, suggesting that the collagen anchoring points would be further apart. We then changed collagen crosslink concentration and used hydrogel-nanoparticle substrates to vary anchoring distance at constant substrate stiffness. Lower collagen anchoring density resulted in increased differentiation. We conclude that stem cells exert a mechanical force on collagen fibres and gauge the feedback to make cell-fate decisions.