The SCD – Stem Cell Differentiation ESA Project: Preparatory Work for the Spaceflight Mission

Due to spaceflight, astronauts experience serious, weightlessness-induced bone loss because of an unbalanced process of bone remodeling that involves bone marrow mesenchymal stem cells (BMSCs), as well as osteoblasts, osteocytes, and osteoclasts. The effects of microgravity on osteo-cells have been extensively studied, but it is only recently that consideration has been given to the role of BMSCs. Previous researches indicated that human BMSCs cultured in simulated microgravity (sim-μg) alter their proliferation and differentiation. The spaceflight opportunities for biomedical experiments are rare and suffer from a number of operative constraints that could bias the validity of the experiment itself, but remain a unique opportunity to confirm and explain the effects due to microgravity, that are only partially activated/detectable in simulated conditions. For this reason, we carefully prepared the SCD – STEM CELLS DIFFERENTIATION experiment, selected by the European Space Agency (ESA) and now on the International Space Station (ISS). Here we present the preparatory studies performed on ground to adapt the project to the spaceflight constraints in terms of culture conditions, fixation and storage of human BMSCs in space aiming at satisfying the biological requirements mandatory to retrieve suitable samples for post-flight analyses. We expect to understand better the molecular mechanisms governing human BMSC growth and differentiation hoping to outline new countermeasures against astronaut bone loss.     

Dynamics of Plug Formation in a Circular Cylinder Under Low Bond Number Conditions: Experiment and Simulation

The goal of the current study is to investigate the dynamics of two phase interface under a low Bond number condition. Silicone oil is injected into a cylinder under a Bond number of about 0.47 via a side tube forming a T-junction with the former. The time evolution of the interface of silicon oil in a cylinder is captured using a high speed camera. The volume at which the plug is formed is then determined using an image processing tool to analyze the captured images. A numerical simulation is carried out where fluid is injected into a cylinder, under a less than unity Bond number condition, via a side tube. Numerical and experimental results are then compared.     

Effects of temperature and dispersants on the phases and morphology of Ag–Cu nanoparticles

Ultrafine Ag–Cu nanoparticles (NPs) have been synthesized by a rapid one-step reduction within only 10 min. Effects of temperature and dispersants on the phases and morphology of Ag–Cu NPs were investigated. Results showed that citric acid exhibited an advantageous nature to avoid the formation of Cu₂O and form uniform morphology over PVP. The average particle size of the Ag–Cu NPs synthesized simply in ice-cubes bath could be controlled in 8.6 nm about a quarter of that synthesized at room temperature. The synthesized Ag–Cu NPs presented alloy states near the eutectic composition of 72:28. Due to the lower Ostwald ripening rate and citric acid protection, smaller Ag–Cu NPs were achieved in ice-cube bath. Results also showed that the ultrafine Ag–Cu NPs could be expected to sinter at about 330 °C which was much lower than the eutectic temperature (779 °C) of bulk Ag–Cu alloy. The ultrafine Ag–Cu NPs could be applied as potential die attach materials for SiC power devices.     

Elastic and viscoelastic mechanical properties of brain tissues on the implanting trajectory of sub-thalamic nucleus stimulation

Corresponding to pre-puncture and post-puncture insertion, elastic and viscoelastic mechanical properties of brain tissues on the implanting trajectory of sub-thalamic nucleus stimulation are investigated, respectively. Elastic mechanical properties in pre-puncture are investigated through pre-puncture needle insertion experiments using whole porcine brains. A linear polynomial and a second order polynomial are fitted to the average insertion force in pre-puncture. The Young’s modulus in pre-puncture is calculated from the slope of the two fittings. Viscoelastic mechanical properties of brain tissues in post-puncture insertion are investigated through indentation stress relaxation tests for six interested regions along a planned trajectory. A linear viscoelastic model with a Prony series approximation is fitted to the average load trace of each region using Boltzmann hereditary integral. Shear relaxation moduli of each region are calculated using the parameters of the Prony series approximation. The results show that, in pre-puncture insertion, needle force almost increases linearly with needle displacement. Both fitting lines can perfectly fit the average insertion force. The Young’s moduli calculated from the slope of the two fittings are worthy of trust to model linearly or nonlinearly instantaneous elastic responses of brain tissues, respectively. In post-puncture insertion, both region and time significantly affect the viscoelastic behaviors. Six tested regions can be classified into three categories in stiffness. Shear relaxation moduli decay dramatically in short time scales but equilibrium is never truly achieved. The regional and temporal viscoelastic mechanical properties in post-puncture insertion are valuable for guiding probe insertion into each region on the implanting trajectory.     

Surface characterization and biological properties of regular dentin,demineralized dentin,and deproteinized dentin

Bone autografts are often used for reconstruction of bone defects; however, due to the limitations of autografts, researchers have been in search of bone substitutes. Dentin is of particular interest for this purpose due to high similarity to bone. This in vitro study sought to assess the surface characteristics and biological properties of dentin samples prepared with different treatments. This study was conducted on regular (RD), demineralized (DemD), and deproteinized (DepD) dentin samples. X-ray diffraction and Fourier transform infrared spectroscopy were used for surface characterization. Samples were immersed in simulated body fluid, and their bioactivity was evaluated under a scanning electron microscope. The methyl thiazol tetrazolium assay, scanning electron microscope analysis and quantitative real-time polymerase chain reaction were performed, respectively to assess viability/proliferation, adhesion/morphology and osteoblast differentiation of cultured human dental pulp stem cells on dentin powders. Of the three dentin samples, DepD showed the highest and RD showed the lowest rate of formation and deposition of hydroxyapatite crystals. Although, the difference in superficial apatite was not significant among samples, functional groups on the surface, however, were more distinct on DepD. At four weeks, hydroxyapatite deposits were noted as needle-shaped accumulations on DemD sample and numerous hexagonal HA deposit masses were seen, covering the surface of DepD. The methyl thiazol tetrazolium, scanning electron microscope, and quantitative real-time polymerase chain reaction analyses during the 10-day cell culture on dentin powders showed the highest cell adhesion and viability and rapid differentiation in DepD. Based on the parameters evaluated in this in vitro study, DepD showed high rate of formation/deposition of hydroxyapatite crystals and adhesion/viability/osteogenic differentiation of human dental pulp stem cells, which may support its osteoinductive/osteoconductive potential for bone regeneration.     

A superhydrophilic titanium implant functionalized by ozone gas modulates bone marrow cell and macrophage responses

Bone-forming cells and M? play key roles in bone tissue repair. In this study, we prepared a superhydrophilic titanium implant functionalized by ozone gas to modulate osteoconductivity and inhibit inflammatory response towards titanium implants. After 24 h of ozone gas treatment, the water contact angle of the titanium surface became zero. XPS analysis revealed that hydroxyl groups were greatly increased, but carbon contaminants were largely decreased 24 h after ozone gas functionalization. Also, ozone gas functionalization did not alter titanium surface topography. Superhydrophilic titanium (O₃–Ti) largely increased the aspect ratio, size and perimeter of cells when compared with untreated titanium (unTi). In addition, O₃–Ti facilitated rat bone marrow derived MSCs differentiation and mineralization evidenced by greater ALP activity and bone-like nodule formation. Interestingly, O₃–Ti did not affect RAW264.7 M? proliferation. However, naive RAW264.7 M? cultured on unTi produced a two-fold larger amount of TNFα than that on O₃–Ti. Furthermore, O₃–Ti greatly mitigated proinflammatory cytokine production, including TNFα and IL-6 from LSP-stimulated RAW264.7 M?. These results demonstrated that a superhydrophilic titanium prepared by simple ozone gas functionalization successfully increased MSCs proliferation and differentiation, and mitigated proinflammatory cytokine production from both naive and LPS-stimulated M?. This superhydrophilic surface would be useful as an endosseous implantable biomaterials and as a biomaterial for implantation into other tissues.     

Nonlinearity of bituminous mixtures

This paper presents an experimental characterization of the strain dependency of the complex modulus of bituminous mixtures for strain amplitude levels lower than about \(110~\upmu\mbox{m}/\mbox{m}\). A series of strain amplitude sweep tests are performed at different temperatures (8, 10, 12 and 14°C) and frequencies (0.3, 1, 3 and 10 Hz), during which complex modulus is monitored. For each combination of temperature and frequency, four maximum strain amplitudes are targeted (50, 75, 100 and \(110~\upmu\mbox{m}/\mbox{m}\)). For each of them, two series of 50 loading cycles are applied, respectively at decreasing and increasing strain amplitudes. Before each decreasing strain sweep and after each increasing strain sweep, 5 cycles are performed at constant maximum targeted strain amplitude.Experimental results show that the behavior of the studied material is strain dependent. The norm of the complex modulus decreases and phase angle increases with strain amplitude. Results are presented in Black and Cole–Cole plots, where characteristic directions of nonlinearity can be identified. Both the effects of nonlinearity in terms of the complex modulus variation and of the direction of nonlinearity in Black space seem to validate the time–temperature superposition principle with the same shift factors as for linear viscoelasticity.The comparison between results obtained during increasing and decreasing strain sweeps suggests the existence of another phenomenon occurring during cyclic loading, which appears to systematically induce a decrease of the norm of the complex modulus and an increase of the phase angle, regardless of the type of the strain sweep (increasing or decreasing).     

Quantification of the inherent uncertainty in the relaxation modulus and creep compliance of asphalt mixes

Advanced material characterization of asphalt concrete is essential for realistic and accurate performance prediction of flexible pavements. However, such characterization requires rigorous testing regimes that involve mechanical testing of a large number of laboratory samples at various conditions and set-ups. Advanced measurement instrumentation in addition to meticulous and accurate data analysis and analytical representation are also of high importance. Such steps as well as the heterogeneous nature of asphalt concrete (AC) constitute major factors of inherent variability. Thus, it is imperative to model and quantify the variability of the needed asphalt material’s properties, mainly the linear viscoelastic response functions such as: relaxation modulus, \(E(t)\), and creep compliance, \(D(t)\). The objective of this paper is to characterize the inherent uncertainty of both \(E(t)\) and \(D(t)\) over the time domain of their master curves. This is achieved through a probabilistic framework using Monte Carlo simulations and First Order approximations, utilizing \(E^{*}\) data for six AC mixes with at least eight replicates per mix. The study shows that the inherent variability, presented by the coefficient of variation (COV), in \(E(t)\) and \(D(t)\) is low at small reduced times, and increases with the increase in reduced time. At small reduced times, the COV in \(E(t)\) and \(D(t)\) are similar in magnitude; however, differences become significant at large reduced times. Additionally, the probability distributions and COVs of \(E(t)\) and \(D(t)\) are mix dependent. Finally, a case study is considered in which the inherent uncertainty in \(D(t)\) is forward propagated to assess the effect of variability on the predicted number of cycles to fatigue failure of an asphalt mix.