This paper reports the synthesis of borosilicate bioactive glasses nanoparticles containing ions of interest (calcium, strontium or silver ions) for the purpose of bone tissue regeneration using a modified Stöber method and their shaping by robocasting to produce macroporous grid-like architectures with controlled interconnected porosity. Main focus is on the design of a paste formulation, the optimization of the robocasting process to produce 3D scaffolds and ion release from the sintered scaffold in solution. Pastes were developed (40 vol% powder loading), characterized, and extruded layer-by-layer to form three-dimensional scaffolds with a grid-like macrostructure (400 μm diameter of the rods). After drying under controlled atmosphere, scaffolds were sintered. The sintering study revealed that the addition of silver reduced the densification point of the borosilicate glass. After immersion in a buffer solution, ionic release (Si, B and Sr) of the sintered scaffold was evaluated, and compared with the synthetized powder. The shaping process did not affect Sr dissolution. This controllable ion-release behavior of the scaffolds are promising for their therapeutic application.

This article aims to investigate the Ultra-High Frequency Dielectrophoresis technique to characterize populations of human mesenchymal stem cells. The principle is based on studying the displacement of a cell in an electric field gradient to quantify a characteristic frequency that establishes the UHF dielectrophoretic 'signature' of the investigated cell population. Preliminary results obtained from patient-derived cultures, in correlation with conventional biological analyses, seem to indicate that the osteogenic differentiation of mesenchymal stem cells can be detected by UHF-DEP, making this technique an interesting approach to characterize hMSC differentiation in the context of optimizing bioceramics for bone regenerative medicine.

The solid-state reaction between a stoichiometric hydroxyapatite (HA) and CuO at temperatures above 1100 degrees C produces pure Cu x -HA phases for x <= 0.7 with the general formula Ca10CuI x (PO4)6(OH)2-x O x . The Cu atoms are located at the center of the hexagonal tunnels between two hydroxyl ligands, as determined by Fourier analysis based on XRD data. During heat treatment, the reduction of Cu2+ ions into Cu+ is concomitant with the stabilization of copper in HA in the hexagonal tunnel. The incorporation of monovalent copper within the apatite, as revealed by XANES spectroscopy, explains the violet color of the samples. The incorporation of Cu+ ions, by substitution of a hydrogen atom by copper(I), results in the formation of linear O-Cu-O chains where the majority of which are isolated for x <= 0.3. In addition, the EXAFS investigation showed, thanks to the linear geometry of these clusters that results in multiple diffusion effects, the existence of [CuO] n chains with n >= 2, which only appear clearly for higher copper contents x >= 0.5. The strong covalency of the Cu-O bond in such a dumbbell configuration would lead to strong hybridization between the 3d and 4s orbitals of copper and the 2p orbitals of oxygen, as illustrated by ESR signals. In the case of Cu-doped HA prepared by coprecipitation and annealed at a lower temperature (T <= 600 degrees C), copper substitutes calcium according to the theoretical formula Ca10-x Cu x (PO4)6(OH)2, mainly at the Ca(2) site. This local environment is in line with the Jahn-Teller distortion induced by the Cu2+ ion (as evidenced by UV-vis-NIR, XPS, and XANES-EXAFS spectroscopy analyses) and also allows copper-copper interactions from one site to another, as observed by ESR spectroscopy. This versatility of copper in HA gives it optical properties that change from a violet color with near-IR absorption to a blue hue. In all cases, Cu-O-Cu interactions persist whatever the valence state, and heat treatment induces a redox phenomenon, with copper exchanging between two sites close to each other.

First posted December 29, 2023 For additional information, contact: Volcano Science Center - Menlo ParkU.S. Geological Survey345 Middlefield Road, MS 910Menlo Park, CA 94025Contact Pubs Warehouse Harrat Rahat, one of several large, basalt-dominated volcanic fields in the western part of the Kingdom of Saudi Arabia, is a prime example of continental, intraplate volcanism. Excellent exposure makes this an outstanding site to investigate changing volcanic flux and composition through time. We present 93 40Ar/39Ar ages and 6 36Cl surface-exposure ages for volcanic deposits throughout northern Harrat Rahat that, integrated with a new geologic map, define 12 eruptive stages. Exposed volcanic deposits in the study area erupted less than 1.2 million years ago (Ma), and 214 of 234 identified eruptions occurred less than 570 thousand years ago (ka). Two eruptions were in the Holocene, including a historically described basaltic eruption in 1256 C.E. and a trachyte eruption newly recognized as Holocene (4.2±5.2 ka). An estimated approximately 82 cubic kilometers (km3; dense rock equivalent) of volcanic products can be documented as having erupted since 1.2 Ma, though this is a lower limit because of concealment of deposits older than 570 ka. Over the last 570 thousand years (k.y.), the average eruption rate was 0.14 cubic kilometers per thousand years (km3/k.y.), but volcanism was episodic with periods alternating between low (0.04–0.06 km3/k.y.) and high (0.1–0.3 km3/k.y.) effusion rates. Before 180 ka, eruptions vented from the volcanic field's dominant eastern vent axis and from a subsidiary, diffuse, western vent axis. After 180 ka, volcanism focused along the eastern vent axis, and the composition of volcanism varied systematically along its length from basalt dominated in the north to trachyte dominated in the south. We hypothesize that these compositional variations younger than 180 k.y. reflect the growth of a mafic intrusive complex beneath the southern part of the vent axis, which led to the development of evolved magmas. Lastly, these new age data allow for a reassessment of the volcanic recurrence interval at northern Harrat Rahat. Based on available data, volcanism in northern Harrat Rahat over the last 180 k.y. is poorly described using a Poisson distribution with a single recurrence interval. Instead, data for northern Harrat Rahat are better described using a mixed exponential distribution that is applicable for volcanic systems characterized by two different eruptive states, where one state with a longer recurrence interval corresponding to periods of low eruption frequency and one state with a shorter recurrence interval corresponding to periods of high eruption frequency. The preferred model for northern Harrat Rahat over the last 180 k.y. uses a long recurrence interval of 4.0 k.y. and a short recurrence interval of 0.22 k.y.

Incorporation of silicate ions in calcium phosphate ceramics (CPC) and modification of their multiscale architecture are two strategies for improving the vascularization of scaffolds for bone regenerative medicine. The response of endothelial cells, actors for vascularization, to the chemical and physical cues of biomaterial surfaces is little documented, although essential. We aimed to characterize in vitro the response of an endothelial cell line, C166, cultivated on the surface CPCs varying either in terms of their chemistry (pure versus silicon-doped HA) or their microstructure (dense versus microporous). Adhesion, metabolic activity, and proliferation were significantly altered on microporous ceramics, but the secretion of the pro-angiogenic VEGF-A increased from 262 to 386 pg/mL on porous compared to dense silicon-doped HA ceramics after 168 h. A tubulogenesis assay was set up directly on the ceramics. Two configurations were designed for discriminating the influence of the chemistry from that of the surface physical properties. The formation of tubule-like structures was qualitatively more frequent on dense ceramics. Microporous ceramics induced calcium depletion in the culture medium (from 2 down to 0.5 mmol/L), which is deleterious for C166. Importantly, this effect might be associated with the in vitro static cell culture. No influence of silicon doping of HA on C166 behavior was detected.

First posted December 29, 2023 For additional information, contact: Volcano Science Center - Menlo ParkU.S. Geological Survey345 Middlefield Road, MS 910Menlo Park, CA 94025 Paleomagnetic rock samples were collected from 173 drill sites in the Quaternary alkali basaltic volcanic field of northern Harrat Rahat, Kingdom of Saudi Arabia. Laboratory measurements on these samples established that lava flows and vent complexes—identified and mapped from field characteristics, rock types, and compositions as products of single or temporally close eruptions—typically record single, or very similar, directions of remanent magnetization. Correlations defined through geologic mapping, spatial association, geochemistry, geochronology, and identical mean remanent directions indicate at least 16 brief episodes of temporally clustered eruptions. These episodes had durations of a few centuries or less. Anomalous remanent magnetic directions were found for at least 13 mapped lavas of northern Harrat Rahat, which demonstrate that they were acquired during brief geomagnetic cryptochrons during the Brunhes Normal Polarity Chron. These uncharacteristic directions enhance the opportunity to identify common eruptive episodes, and to better understand and evaluate assessments of eruption ages based on 40Ar/39Ar geochronology. Combining paleomagnetic and regional archaeomagnetic results for the youngest eruptions allows us to evaluate their historical age assignments and, in one case, refute a previously assigned provisional age.

The purpose of the study was to investigate the synthesis of economic calcium phosphate powders from recycled oyster shells, using a ball milling method. The oyster shell powder and a calcium pyrophosphate powder were used as starting materials and ball milled, then heat treated at 1,050°C for 5 h to produce calcium phosphate powders through a solid-state reaction. Electrochemically synthesized mesoporous silicon microparticles were then added to the prepared phosphate powders by mechanical mixer. The final powders were characterized using X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy to analyze their chemical composition and determine the most suitable process conditions. The biocompatibility of the produced powders was also tested in vitro using murine cells and the results showed good biocompatibility.

First posted December 29, 2023 For additional information, contact: Volcano Science Center - Menlo ParkU.S. Geological Survey345 Middlefield Road, MS 910Menlo Park, CA 94025Contact Pubs Warehouse The northernmost part of the Harrat Rahat volcanic field contains early Pleistocene to Holocene mafic eruptive products within the vicinity of the city of Al Madīnah, Kingdom of Saudi Arabia. A detailed geologic investigation into the eruptive history of a 570 square kilometer (km2) area covering Al Madīnah and the surrounding area has yielded 33 mapped Quaternary volcanic units consisting of lava flows, scoria cones, and shield volcanoes. These eruptive products consist of continental, intraplate alkalic and minor transitional basalts, hawaiites, and a single mugearite that were emplaced from at least 1,014±14 thousand years ago (ka) to a single Holocene eruption in 1256 C.E. Lava flows are generally 10 to 15 kilometers (km) long (but can reach 23 km long), 1 to 3 km wide, and at least 10 meters thick. Most of the mapped units erupted episodically between 400 and 340 ka and 180 and 100 ka. Despite small individual volumes (less than 1 cubic kilometers dense rock equivalent), each unit represents eruption of a distinct magma batch that was strongly influenced by clinopyroxene, olivine, and plagioclase fractionation. Some of these units are interpreted to have undergone magma mixing pre- and (or) syneruptively. Integrating eruption ages, geochemistry, and paleomagnetic data yields evidence that some eruptions were temporally and (or) spatially clustered. Aligned scoria cones and elongate vent edifices were constructed atop fissure vent systems that reflect the local stress field, which controls dike ascent through the middle and upper crust.

First posted December 29, 2023 For additional information, contact: Volcano Science Center - Menlo ParkU.S. Geological Survey345 Middlefield Road, MS 910Menlo Park, CA 94025 Harrat Rahat is an alkali basalt, continental, intraplate volcanic field located within the central-western part of the Kingdom of Saudi Arabia. The northern quarter of Harrat Rahat contains evolved volcanic products that achieve trachyte compositions (>60 weight percent SiO2). Within the Al Efairia volcanic center, pyroclastic-flow and -surge deposits that reflect explosive trachyte volcanism (and minor exposed lava domes that reflect effusive trachyte eruptions) sit at the surface as the youngest expression of volcanic activity within this part of the Harrat Rahat volcanic field. Five trachyte deposits emplaced explosively have been identified within the Al Efairia volcanic center based on geologic mapping, petrography, geochemistry, and paleomagnetism. These units are the trachytes of Um Rgaibah, Gura 5, Gura 4, Al Efairia, and Al Qayf, in descending stratigraphic order. Here, we present 14 40Ar/39Ar analyses from four of these units, which yield eruption ages of 4.2±5.2 thousand years (ka) for the trachyte of Um Rgaibah, 79.7±1.6 ka for the trachyte of Gura 5, 84.3±1.6 ka for the trachyte of Gura 4, and 88.0±1.8 ka for the trachyte of Al Efairia. The eruption age of the trachyte of Al Qayf has been constrained to between 410.3±3.4 and 418.8±1.9 ka using paleomagnetic correlations and 40Ar/39Ar ages from overlying and underlying intermediate composition lava flows. Most of these trachytes have distinct geochemical compositions, petrographic characteristics, and directions of remanent magnetization. The exceptions are for the trachytes of Gura 4 and Gura 5, which overlap in their geochemical, petrographic, paleomagnetic, and geochronologic affinities. Based on these similarities, we interpret the trachytes of Gura 4 and Gura 5 to have erupted during a closely spaced (a few decades) time interval from the same magma batch but from craters that are >2 kilometers (km) apart. The eruption of the trachyte of Al Efairia at 88.0±1.8 ka is the result of a different magma batch that erupted a few thousand years prior to the trachytes of Gura 4 and Gura 5. The Al Efairia volcanic center is remarkably different from the Matan volcanic center located ~10 km to the north, which has also erupted young (<150 ka) trachytes. The Matan volcanic center has been shown to produce trachyte compositions only after eruption of basalt followed by intermediate lava flows, whereas only trachyte compositions have erupted within the Al Efairia volcanic center over this same time interval.

At least 28 rhyolitic lava flows, domes, and tuffs erupted within Yellowstone caldera following its formation 631 ka. Understanding the timing of intracaldera eruptions is essential for characterizing natural hazards posed by Yellowstone volcano. We present 40 Ar/ 39 Ar eruption ages for the Mallard Lake Member and Central Plateau Member of the Plateau Rhyolite, which comprise products from 22 eruptions spanning ~160 to ~70 ka. These eruptions occurred along two linear vent zones within Yellowstone caldera that appear to be extensions of major extracaldera normal faults. Eruptions took place in five brief clusters with group mean ages of 160.3±1.0 ka (2σ), 149.8±4.0 ka, 111.4±0.9 ka, 104.1±0.8 ka, and 70.8±0.7 ka. All mapped products within each informal group have indistinguishable 40 Ar/ 39 Ar eruption ages implying brief eruption durations. Eruption products of each group are spatially clustered, with only one of the two linear vent zones being active during an eruption group. Using the oldest group (160 ka), we apply paleomagnetic and geochemical analyses to investigate the duration of these eruption groups and their pre-eruptive magma reservoir configuration. Paleomagnetic analyses suggest that the nine rhyolite eruptions ~160 ka spanned 400 years or less. Glass and sanidine compositions of each rhyolite are nearly identical, which is consistent with pre-eruptive storage of a large (~130 km 3 ), interconnected melt body within Yellowstone volcano’s crystal-rich magma reservoir. These characteristics differ from intracaldera eruptions that vented near the caldera’s inner ring fracture system, which tend to be compositionally disparate with less temporal clustering. These insights inform hazard mitigation scenarios.

Coherent Raman imaging has been extensively applied to live-cell imaging in the last 2 decades, allowing to probe the intracellular lipid, protein, nucleic acid, and water content with a high-acquisition rate and sensitivity. In this context, multiplex coherent anti-Stokes Raman scattering (MCARS) microspectroscopy using sub-nanosecond laser pulses is now recognized as a mature and straightforward technology for label-free bioimaging, offering the high spectral resolution of conventional Raman spectroscopy with reduced acquisition time. Here, we introduce the combination of the MCARS imaging technique with unsupervised data analysis based on multivariate curve resolution (MCR). The MCR process is implemented under the classical signal non-negativity constraint and, even more originally, under a new spatial constraint based on cell segmentation. We thus introduce a new methodology for hyperspectral cell imaging and segmentation, based on a simple, unsupervised workflow without any spectrum-to-spectrum phase retrieval computation. We first assess the robustness of our approach by considering cells of different types, namely, from the human HEK293 and murine C2C12 lines. To evaluate its applicability over a broader range, we then study HEK293 cells in different physiological states and experimental situations. Specifically, we compare an interphasic cell with a mitotic (prophase) one. We also present a comparison between a fixed cell and a living cell, in order to visualize the potential changes induced by the fixation protocol in cellular architecture. Next, with the aim of assessing more precisely the sensitivity of our approach, we study HEK293 living cells overexpressing tropomyosin-related kinase B (TrkB), a cancer-related membrane receptor, depending on the presence of its ligand, brain-derived neurotrophic factor (BDNF). Finally, the segmentation capability of the approach is evaluated in the case of a single cell and also by considering cell clusters of various sizes.

Hydroxyapatite (HA) ceramic scaffolds are commonly used as bone graft substitutes. Design of such scaffolds is a challenge to improve biological properties and extend the applications of HA ceramics in the field of bone tissue engineering. In this work, we investigated the processing and the in vitro properties of HA ceramic scaffolds mimicking human trabecular bone architecture. Samples of human tibial trabecular bone were collected (University Hospital Center of Limoges) and scanned by X-Ray μ-computed tomography (μ-CT) to generate 3D model database. From this computer-aided design, HA ceramic scaffolds were shaped layer-by-layer by additive manufacturing using laser stereolithography (SLA). Then, green parts were sintered to obtain dense ceramic scaffolds. The shaped parts were compared to the model (wall thickness, size, and geometry of the porous network) using image analysis. A good agreement was found. Only small differences were detected due to a light overpolymerization or to some unprinted very small details that were not linked to a polymerized area of the previous layer. Due to part shrinkage during sintering a magnifying factor has to be applied to the scanned CAO model to match the real dimensions of the trabecular bone sample. Human mesenchymal stem cell (hMSC) cultures were performed to investigate the biological properties of these scaffolds (cell attachment and proliferation of hMSC). These preliminary biological evaluations show the good biocompatibility and cell adhesion of the HA substitute. This work evidences the efficiency of SLA to produce ceramic scaffold architectures mimicking that of the natural trabecular bone with promising biological behavior.

The aim of this work was to identify robust and reproducible signatures characterizing the different steps of bone cell differentiation, from precursors to mature bone cells, using approaches allowing characterization by label-free imaging. Human mesenchymal stromal cells (hMSCs) were cultured either in a growth medium (GM), unable to induce cell differentiation by itself, or in an osteogenic differentiation medium (ODM) on hydroxyapatite ceramics or borosilicate glass. Cell density as well as cell structure, size, and morphology were investigated. A fluorescence microscopy-based approach was followed, using fluorescent labelling of cell features. Some early morphological changes of hMSC during osteogenic differentiation were identified as soon as 48h that were accentuated after 7 days of culture. Cell density was higher when cells were cultured in GM and the cells exhibited significantly smaller nuclei (size ratio about 1.3-1.5) than those cultured in ODM, regardless of the culture support. In ODM, the cells were also of bigger size (1.2 to 1.5 times) and their focal adhesions were reinforcedType I collagen, a gold standard marker of osteogenic differentiation, appeared more intense in ODM. These cell features can be determined using multimodal label-free imaging methods to characterize the differentiation state of hMSCs at the biomaterial surface. They give rise to new cost-effective approaches to investigate cell behavior by suppressing the chemical markers and reducing both the number of needed samples and the requested time to do so.

Les propriétés biologiques des céramiques phosphocalciques, dont l’hydroxyapatite, de composition chimique proche de celle de l’os minéral ne sont pas suffisantes pour répondre à l’ensemble des besoins en médecine régénérative osseuse. Pour en améliorer les performances biologiques plusieurs leviers d’actions existent à différents niveaux de la chaîne d’élaboration de ces biomatériaux. Or, chaque maillon de cette chaîne correspond à un champ disciplinaire spécifique lié à un domaine scientifique différent (ex : chimie des matériaux vs. biologie). L’organisation de la recherche menée à l’IRCER autour de cette thématique est donc le reflet de cette transdisciplinarité. Elle implique une perméabilité et un dialogue constant entre les disciplines et permet la maîtrise de la recherche depuis la synthèse du matériau jusqu’à l’évaluation de ses performances biologiques.

In this fundamental solid-state chemistry study, two sample series were investigated in depth: iron(III)-doped hydroxyapatite (HA) compounds obtained from a co-sintering process of hematite and pure HA under air and iron(III)-doped HA compounds obtained from a co-sintering process from iron(II) acetate and pure HA under an argon atmosphere. X-ray diffraction, UV-visible, Fourier transform infrared, 1H and 31P NMR, electron paramagnetic resonance (EPR,) and Mössbauer spectroscopy methods were coupled to unravel the Fe valence states, the interactions with other anionic species (OH- and PO43-), and finally the complex local environments in hexagonal channels in both the series. In particular, we highlighted the associated mechanism to ensure electroneutrality with a focus on deprotonation versus calcium substitution. By diverging mechanisms, Fe3+ and Fe2+ ions were found to be located in different coordinated sites: 4(+1) coordinated site for Fe3+ and 2(+3) coordinated site for Fe2+ and clearly associated with very different Mössbauer and EPR signatures as various absorption bands (leading to different sample colors).

Implants made of ceramics, and more particularly of calcium phosphates (hydroxyapatite: HA, mainly), promoting intimate contact with natural bone are nowadays merging. Addition of copper ions in bio-ceramics is expected to increase the biological compatibilities of bone graft substitutes. Previous works have shown that copper-doped hydroxyapatite (Cu-doped HA) ceramics can be prepared by solid-state sintering between HA and CuO powder mixtures at about 1100 degrees C; but, copper-substituted HA was found to be metastable leading to apatitic grains and Cu-rich grain boundaries during the sintering process. Ultra-rapid sintering is so needed. Selective laser sintering (SLS) is an additive manufacturing process that possesses the advantage to be based on ultra-fast sintering process under laser irradiation. SLS being used in literature for the application of laser on polymer-ceramic or poymer-metal composites the proper term all along the paper is Ceramic Powder Bed Laser Sintering (CPBLS). To achieve densification of Cu-doped HA ceramics from CPBLS process, one should control the composition/morphology/structure of the powder bed as well as three other important CPBLS parameters: (i) the applied energy from the laser beam, (ii) the laser power and the laser scanning speed, (iii) the distance between two successive lased lines. In this paper, the impact of all the main CPBLS parameters controlling the sintering of dip-coated Cu-doped HA layers on glass substrates is carefully investigated. Possibility of the creation of thin consolidated Cu-doped HA ceramic patterns, using the ultra-fast CPBLS process, is finally shown.

Coherent anti-Stokes Raman scattering (CARS) is a vibrational spectroscopy method based on a coherent χ (3) nonlinear interaction between light and matter through four-wave mixing. CARS process has been extensively used as a microscopy technique in biology. However the shape of CARS spectra is generally complex because of the combination of both resonant and non-resonant contributions, which limits the development of CARS microspectroscopy applications. In this context, phase retrieval methods (PRM) like the maximum entropy method (MEM) [1] are commonly used to extract the vibrationally resonant signal, corresponding to Im[χ (3) ], and thus to provide Raman-like spectra, which are more suitable for interpretation.