The application of heat‐deproteinization to the morphological study of cortical bone: A contribution to the knowledge of the osteonal structure

Observation of heat‐deproteinized cortical bone specimens in incident light enabled the high definition documentation of the osteonal pattern of diaphyseal Haversian bone. This prompted a study to compare these images with those revealed by polarized light microscopy, carried out either on decalcified or thin, undecalcified, resin‐embedded sections. Different bone processing methods can reveal structural aspects of the intercellular matrix, depending on the light diffraction mode: birefringency in decalcified sections can be ascribed to the collagen fibrils orientation alone; in undecalcified sections, to both the ordered layout of collagen and the inorganic phase; in the heat‐deproteinized samples, exclusively to the hydroxyapatite crystals aggregation mode. The elemental chemical analysis documented low content of carbon and hydrogen, no detectable levels of nitrogen and significantly higher content of calcium and phosphorus in heat‐deproteinized samples, as compared with dehydrated controls. In both samples, the X‐ray diffraction (XRD) pattern did not show any significant difference in pattern of hydroxyapatite, with no peaks of any possible decomposition phases. Scanning electron microscopic (SEM) morphology of heat‐deproteinized samples could be documented with the fracturing technique facilitated by the bone brittleness. The structure of crystal aggregates, oriented in parallel and with marks of time periods, was documented. Comparative study of deproteinized and undecalcified samples showed that the matrix inorganic phase did not undergo a coarse grain thermal conversion until it reached 500°C, maintaining the original crystals structure and orientation. Incident light stereomicroscopy, combined with SEM analysis of deproteinized bone fractured surfaces, is a new enforceable technique which can be used in morphometric studies to improve the understanding of the osteonal dynamics. Microsc. Res. Tech. 79:691–699, 2016. © 2016 Wiley Periodicals, Inc.     

Description of a propulsion unit used in guiding a walking machine by recognizing a three-point bordered path

A reconfigurable propulsion unit based on the Peaucellier-Lipkin mechanism has the ability to describe exact straight or curved paths depending on the selected ratio between the lengths of two of its links. The Peaucellier-Lipkin mechanism with one degree of freedom is transformed into a more sophisticated parallel kinematic chain by including four more degrees of freedom. The resulting propulsion unit is able to adapt its kinematic structure and reach instant centers of rotation, in accordance with the presence of three points that border a geometric path. A laser sensor mounted on the body of the machine detects each point. Once the machine has detected the exact location of the border of the road, it walks along a curve parallel to that border. Although the proposed research describes only one propulsion unit or leg, the methodology can be applied to all the legs of the walking machine. The novel 5-DOF leg is able to reach different centers of rotation, providing either the concave or convex arcs that satisfy the basic principle of displacement of walking machines.     

A Robust Design for Cellular Vehicles of Gold Nanorods for Multimodal Imaging

The pursuit of more selectivity in the delivery of plasmonic particles to tumors is critical before their penetration into clinical applications as the photoacoustic imaging and the photothermal ablation of cancer. As their direct infusion into the bloodstream remains problematic, due to a multitude of biological barriers, the development of alternative approaches is emerging as a new challenge. In this context, the recruitment of homologous tumor‐tropic cells that may serve as Trojan horses stands out as a fascinating possibility. Here, a novel model of gold nanorods is presented that feature a composite shell and undergo efficient and reproducible endocytic uptake from murine macrophages, which is fine‐tunable over a broad range of conditions. These cells preserve their viability and more than 90% of their innate chemotactic behavior in vitro, even with a cargo exceeding 200 000 particles per cell. In addition, we show that these vehicles are detectible by photoacoustic imaging down to concentrations in the order of 1% in whole blood and by clinical X‐ray computed tomography below 10%, which is within the typical fraction of a leukocytic infiltrate in a tumor microenvironment, and may even work as contrast agents for the photothermal ablation of cancer.     

Integrin-targeting with peptide-bioconjugated semiconductor-magnetic nanocrystalline heterostructures

Binary asymmetric nanocrystals (BNCs), composed of a photoactive TiO₂ nanorod joined with a superparamagnetic γ-Fe₂O₃ spherical domain, were embedded in polyethylene glycol modified phospholipid micelle and successfully bioconjugated to a suitably designed peptide containing an RGD motif. BNCs represent a relevant multifunctional nanomaterial, owing to the coexistence of two distinct domains in one particle, characterized by high photoactivity and magnetic properties, that is particularly suited for use as a phototherapy and hyperthermia agent as well as a magnetic probe in biological imaging. We selected the RGD motif in order to target integrin expressed on activated endothelial cells and several types of cancer cells. The prepared RGD-peptide/BNC conjugates, comprehensively monitored by using complementary optical and structural techniques, demonstrated a high stability and uniform dispersibility in biological media. The cytotoxicity of the RGD-peptide/BNC conjugates was studied in vitro. The cellular uptake of RGD-peptide conjugates in the cells, assessed by means of two distinct approaches, namely confocal microscopy analysis and emission spectroscopy determination in cell lysates, displayed selectivity of the RGD-peptide-BNC conjugate for the αvβ3 integrin. These RGD-peptide-BNC conjugates have a high potential for theranostic treatment of cancer.

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Dry coating of solid dosage forms: an overview of processes and applications

Dry coating techniques enable manufacturing of coated solid dosage forms with no, or very limited, use of solvents. As a result, major drawbacks associated with both organic solvents and aqueous coating systems can be overcome, such as toxicological, environmental, and safety-related issues on the one hand as well as costly drying phases and impaired product stability on the other. The considerable advantages related to solventless coating has been prompting a strong research interest in this field of pharmaceutics. In the article, processes and applications relevant to techniques intended for dry coating are analyzed and reviewed. Based on the physical state of the coat-forming agents, liquid- and solid-based techniques are distinguished. The former include hot-melt coating and coating by photocuring, while the latter encompass press coating and powder coating. Moreover, solventless techniques, such as injection molding and three-dimensional printing by fused deposition modeling, which are not purposely conceived for coating, are also discussed in that they would open new perspectives in the manufacturing of coated-like dosage forms.     

Modeled Microgravity Affects Fibroblast Functions Related to Wound Healing

Wound healing is crucial for the survival of an organism. Therefore, in the perspective of space exploration missions, it is important to understand if and how microgravity conditions affect the behavior of the cell populations involved in wound healing and the evolution of the process. Since fibroblasts are the major players in tissue repair, this study was focused on the behavior of fibroblasts in microgravity conditions, modeled by a RCCS. Cell cytoskeleton was studied by immunofluorescence microscopy, the ability to migrate was assessed by microchemotaxis and scratch assay, and the expression of markers of fibroblast activation, angiogenesis, and inflammation was assessed by western blot. Results revealed that after cell exposure to modeled microgravity conditions, a thorough rearrangement of microtubules occurred and α-SMA bundles were replaced by a tight network of faulty and disorganized filaments. Exposure to modeled microgravity induced a decrease in α-SMA and E-CAD expressions. Also, the expression of the pro-angiogenic protein VEGF decreased, while that of the inflammatory signal COX-2 increased. Fibroblast ability to adhere, migrate, and respond to chemoattractants (PRP), closely related to cytoskeleton integrity and membrane junctions, was significantly impaired. Nevertheless, PRP was able to partially restore fibroblast migration.     

Mixed Fluorinated/Hydrogenated Self‐Assembled Monolayer‐Protected Gold Nanoparticles: In Silico and In Vitro Behavior

Gold nanoparticles (AuNPs) covered with mixtures of immiscible ligands present potentially anisotropic surfaces that can modulate their interactions at complex nano–bio interfaces. Mixed, self‐assembled, monolayer (SAM)‐protected AuNPs, prepared with incompatible hydrocarbon and fluorocarbon amphiphilic ligands, are used here to probe the molecular basis of surface phase separation and disclose the role of fluorinated ligands on the interaction with lipid model membranes and cells, by integrating in silico and experimental approaches. These results indicate that the presence of fluorinated amphiphilic ligands enhances the membrane binding ability and cellular uptake of gold nanoparticles with respect to those coated only with hydrogenated amphiphilic ligands. For mixed monolayers, computational results suggest that ligand phase separation occurs on the gold surface, and the resulting anisotropy affects the number of contacts and adhesion energies with a membrane bilayer. This reflects in a diverse membrane interaction for NPs with different surface morphologies, as determined by surface plasmon resonance, as well as differential effects on cells, as observed by flow cytometry and confocal microscopy. Overall, limited changes in monolayer features can significantly affect NP surface interfacial properties, which, in turn, affect the interaction of SAM‐AuNPs with cellular membranes and subsequent effects on cells.     

High‐Density Sb2Te3 Nanopillars Arrays by Templated,Bottom‐Up MOCVD Growth

Sb₂Te₃ exhibits several technologically relevant properties, such as high thermoelectric efficiency, topological insulator character, and phase change memory behavior. Improved performances are observed and novel effects are predicted for this and other chalcogenide alloys when synthetized in the form of high‐aspect‐ratio nanostructures. The ability to grow chalcogenide nanowires and nanopillars (NPs) with high crystal quality in a controlled fashion, in terms of their size and position, can boost the realization of novel thermoelectric, spintronic, and memory devices. Here, it is shown that highly dense arrays of ultrascaled Sb₂Te₃ NPs can be grown by metal organic chemical vapor deposition (MOCVD) on patterned substrates. In particular, crystalline Sb₂Te₃ NPs with a diameter of 20 nm and a height of 200 nm are obtained in Au‐functionalized, anodized aluminum oxide (AAO) templates with a pore density of ≈5 × 10¹⁰ cm^?2. Also, MOCVD growth of Sb₂Te₃ can be followed either by mechanical polishing and chemical etching to produce Sb₂Te₃ NPs arrays with planar surfaces or by chemical dissolution of the AAO templates to obtain freestanding Sb₂Te₃ NPs forests. The illustrated growth method can be further scaled to smaller pore sizes and employed for other MOCVD‐grown chalcogenide alloys and patterned substrates.     

Toward High‐Dimensional Single‐Cell Analysis of Graphene Oxide Biological Impact: Tracking on Immune Cells by Single‐Cell Mass Cytometry

Considering the potential exposure to graphene, the most investigated nanomaterial, the assessment of the impact on human health has become an urgent need. The deep understanding of nanomaterial safety is today possible by high‐throughput single‐cell technologies. Single‐cell mass cytometry (cytometry by time‐of flight, CyTOF) shows an unparalleled ability to phenotypically and functionally profile complex cellular systems, in particular related to the immune system, as recently also proved for graphene impact. The next challenge is to track the graphene distribution at the single‐cell level. Therefore, graphene oxide (GO) is functionalized with AgInS₂ nanocrystals (GO–In), allowing to trace GO immune–cell interactions via the indium (¹¹⁵In) channel. Indium is specifically chosen to avoid overlaps with the commercial panels (>30 immune markers). As a proof of concept, the GO–In CyTOF tracking is performed at the single‐cell level on blood immune subpopulations, showing the GO interaction with monocytes and B cells, therefore guiding future immune studies. The proposed approach can be applied not only to the immune safety assessment of the multitude of graphene physical and chemical parameters, but also for graphene applications in neuroscience. Moreover, this approach can be translated to other 2D emerging materials and will likely advance the understanding of their toxicology.