Simultaneous submicrometric 3D imaging of the micro-vascular network and the neuronal system in a mouse spinal cord

Defaults in vascular (VN) and neuronal networks of spinal cord are responsible for serious neurodegenerative pathologies. Because of inadequate investigation tools, the lacking knowledge of the complete fine structure of VN and neuronal systems is a crucial problem. Conventional 2D imaging yields incomplete spatial coverage leading to possible data misinterpretation, whereas standard 3D computed tomography imaging achieves insufficient resolution and contrast. We show that X-ray high-resolution phase-contrast tomography allows the simultaneous visualization of three-dimensional VN and neuronal systems of mouse spinal cord at scales spanning from millimeters to hundreds of nanometers, with neither contrast agent nor a destructive sample-preparation. We image both the 3D distribution of micro-capillary network and the micrometric nerve fibers, axon-bundles and neuron soma. Our approach is a crucial tool for pre-clinical investigation of neurodegenerative pathologies and spinal-cord-injuries. In particular, it should be an optimal tool to resolve the entangled relationship between VN and neuronal system.Comment: 15 pages, 6 figure

Enhanced and Flexible Software Tools for X-ray Computed Tomography at the Italian Synchrotron Radiation Facility Elettra

X-ray computed tomography (CT) experiments performed at synchrotron radiation facilities require adequate computing and storage resources due to the large amount of acquired and reconstructed data produced. To satisfy the heterogeneous needs of beamline users, flexible solutions are also required. Moreover, the growing demand of quantitative image analysis impose an easy integration between the CT reconstruction process and the subsequent feature extraction step. This paper presents some of the software solutions adopted by the SYRMEP beamline of the Italian synchrotron radiation facility Elettra. By using the enhanced version of the reconstruction software here presented as well as data reduction and data analysis tools, beamline users can easily implement an integrated and comprehensive approach to the digital image processing and image analysis required by a tomography-oriented scientific workflow

Анализ и пути улучшения финансового состояния предприятия (на примере ГЛХУ «Хойникский лесхоз»)

Lung imaging in mouse disease models is crucial for the assessment of the severity of airway disease but remains challenging due to the small size and the high porosity of the organ. Synchrotron inline free-propagation phase-contrast computed tomography (CT) with its intrinsic high soft-tissue contrast provides the necessary sensitivity and spatial resolution to analyse the mouse lung structure in great detail. Here, this technique has been applied in combination with single-distance phase retrieval to quantify alterations of the lung structure in experimental asthma mouse models of different severity. In order to mimic an in vivo situation as close as possible, the lungs were inflated with air at a constant physiological pressure. Entire mice were embedded in agarose gel and imaged using inline free-propagation phase-contrast CT at the SYRMEP beamline (Synchrotron Light Source, Elettra, Trieste, Italy). The quantification of the obtained phase-contrast CT data sets revealed an increasing lung soft-tissue content in mice correlating with the degree of the severity of experimental allergic airways disease. In this way, it was possible to successfully discriminate between healthy controls and mice with either mild or severe allergic airway disease. It is believed that this approach may have the potential to evaluate the efficacy of novel therapeutic strategies that target airway remodelling processes in asthma.Funding Agencies|German Research Foundation (Deutsche Forschungsgemeinschaft, DFG) [DU 1403/1-1]</p

Meniscal ossicles as micro-CT imaging biomarker in a rodent model of antigen-induced arthritis: A synchrotron-based x-ray pilot study

It is increasingly recognized that early detection of bone erosion plays an important role in the overall evaluation of rheumatoid arthritis and in the choice of the correct treatment approach. Since an appropriate use of imaging biomarkers in preclinical settings offers the prospect of smaller and optimized sample size, in the present study we define an anatomical imaging biomarker that could be objectively measured from micro-CT imaging data as an indicator of bone erosion in arthritis process. The well-characterized antigen-induced arthritis (AIA) model in rats was used. The animals were divided into 2 groups: arthritic disease control and arthritic having been administrated with the tumor necrosis factor alpha-blocking agent (Humira). Rats were sacrificed in the acute phase of AIA; peripheral blood and synovial tissue were collected for assessment of arthritis. Ex vivo micro-CT tomography of knee joints was performed at the Elettra synchrotron light source (Trieste, Italy). Overall, results from this study suggest that use of high-resolution micro-CT analysis coupled with meniscal ossicles bone parameters quantification provide a powerful combination to enhance data interpretation and assessment of disease-modifying drugs in an animal model of arthritis

A Modified Rabbit Ulna Defect Model for Evaluating Periosteal Substitutes in Bone Engineering: A Pilot Study

The present work defines a modified critical size rabbit ulna defect model for bone regeneration in which a non-resorbable barrier membrane was used to separate the radius from the ulna to create a valid model for evaluation of tissue-engineered periosteal substitutes. Eight rabbits divided into two groups were used. Critical defects (15 mm) were made in the ulna completely eliminating periosteum. For group I, defects were filled with a nanohydroxyapatite poly(ester urethane) scaffold soaked in PBS and left as such (group Ia) or wrapped with a tissue-engineered periosteal substitute (group Ib). For group II, an expanded-polytetrafluoroethylene (e-PTFE) (GORE-TEX\uae) membrane was inserted around the radius then the defects received either scaffold alone (group IIa) or scaffold wrapped with periosteal substitute (group IIb). Animals were euthanized after 12\u201316 weeks, and bone regeneration was evaluated by radiography, computed microtomography (\ub5CT), and histology. In the first group, we observed formation of radio-ulnar synostosis irrespective of the treatment. This was completely eliminated upon placement of the e-PTFE (GORETEX\uae) membrane in the second group of animals. In conclusion, modification of the model using a non-resorbable e-PTFE membrane to isolate the ulna from the radius was a valuable addition allowing for objective evaluation of the tissue-engineered periosteal substitut

In Vivo Regenerative Properties of Coralline-Derived (Biocoral) Scaffold Grafts in Human Maxillary Defects: Demonstrative and Comparative Study with Beta-Tricalcium Phosphate and Biphasic Calcium Phosphate by Synchrotron Radiation X-Ray Microtomography

Background: In recent years, there has been interest on the fabrication of systems using particulates or block-based approach for bone tissue engineering (TE) scaffolds, possessing porous interconnected structures. In fact, these particular morphologies greatly increase the surface area for more chemical and biological reactions to take place. Purpose: This study was designed to demonstrate the unique capability of the synchrotron radiation x-ray microtomography (micro-CT) in offering an advanced characterization of coralline-derived (Biocoral) biomaterials placed in human maxillary defects as it allows, in a nondestructive way, a complete, precise, and high-resolution three-dimensional analysis of their microstructural parameters. Moreover, the comparison between Biocoral and other biomaterials was explored to understand the mechanism of their biological behavior as bone substitute. Materials and Methods: Implant survival, bone regeneration, graft resorption, neovascularization, and morphometric parameters (including anisotropy and connectivity index of the structures) were evaluated by micro-CT in Biocoral and the other biomaterials after 6 to 7 months from implantation in human maxillary bone defects. Results: After the in vivo tests, a huge amount of bone was detected in the retrieved Biocoral-based samples, coupled with a good rate of biomaterial resorption and the formation of a homogeneous and rich net of new vessels. The morphometric parameters were comparable to those obtained in the biphasic calcium phosphate-based control, with the exception of the connectivity index for which this control exhibited the most well-connected structure. This last result, together with those referred to the poor performances of the b-tricalcium phosphate block-based sample, suggests that the particular scaffold morphology may play a role in the hunt the optimal scaffold structure to be implanted. Conclusion: In this limited study, implant success rate seems not strictly dependent on the biomaterial that is used, but on the scaffold morphology. Micro-CT technique was demonstrated to play a fundamental role in advanced characterization of bone TE constructs

Alpine dwarf shrubs show high proportions of nonfunctional xylem: Visualization and quantification of species-specific patterns

Xylem conductive capacity is a key determinant of plant hydraulic function and intimately linked to photosynthesis and productivity, but can be impeded by temporary or permanent conduit dysfunctions. Here we show that persistent xylem dysfunctions in unstressed plants are frequent in Alpine dwarf shrubs and occur in various but species-specific cross-sectional patterns. Combined synchrotron micro-computed tomography (micro-CT) imaging, xylem staining, and flow measurements in saturated samples of six widespread Ericaceae species evidence a high proportion (19%-50%) of hydraulically nonfunctional xylem areas in the absence of drought stress, with regular distribution of dysfunctions between or within growth rings. Dysfunctions were only partly reversible and reduced the specific hydraulic conductivity to 1.38 to 3.57 ×10-4 m2 s-1 MPa-1 . Decommission of inner growth rings was clearly related to stem age and a higher vulnerability to cavitation of older rings, while the high proportion of nonfunctional conduits in each annual ring needs further investigations. The lower the xylem fraction contributing to the transport function, the higher was the hydraulic efficiency of conducting xylem areas. Improved understanding of the functional lifespan of xylem elements and the prevalence and nature of dysfunctions is critical to correctly assess structure-function relationships and whole-plant hydraulic strategies

Clinical study in phase- contrast mammography: image-quality analysis

The first clinical study of phase-contrast mammography (PCM) with synchrotron radiation was carried out at the Synchrotron Radiation for Medical Physics beamline of the Elettra synchrotron radiation facility in Trieste (Italy) in 2006–2009. The study involved 71 patients with unresolved breast abnormalities after conventional digital mammography and ultrasonography exams carried out at the Radiology Department of Trieste University Hospital. These cases were referred for mammography at the synchrotron radiation facility, with images acquired using a propagation-based phase-contrast imaging technique. To investigate the contribution of phase-contrast effects to the image quality, two experienced radiologists specialized in mammography assessed the visibility of breast abnormalities and of breast glandular structures. The images acquired at the hospital and at the synchrotron radiation facility were compared and graded according to a relative seven-grade visual scoring system. The statistical analysis highlighted that PCM with synchrotron radiation depicts normal structures and abnormal findings with higher image quality with respect to conventional digital mammography

X-ray microtomography observations of xylem embolism in stems of Laurus nobilis are consistent with hydraulic measurements of percentage loss of conductance

Drought-induced xylem embolism is a serious threat to plant survival under future climate scenarios. Hence, accurate quantification of species-specific vulnerability to xylem embolism is a key to predict the impact of climate change on vegetation. Low-cost hydraulic measurements of embolism rate have been suggested to be prone to artefacts, thus requiring validation by direct visualization of the functional status of xylem conduits using nondestructive imaging techniques, such as X-ray microtomography (microCT). We measured the percentage loss of conductance (PLC) of excised stems of Laurus nobilis (laurel) dehydrated to different xylem pressures, and compared results with direct observation of gas-filled vs water-filled conduits at a synchrotron-based microCT facility using a phase contrast imaging modality. Theoretical PLC calculated on the basis of microCT observations in stems of laurel dehydrated to different xylem pressures overall were in agreement with hydraulic measurements, revealing that this species suffers a 50% loss of xylem hydraulic conductance at xylem pressures averaging 3.5 MPa. Our data support the validity of estimates of xylem vulnerability to embolism based on classical hydraulic techniques. We discuss possible causes of discrepancies between data gathered in this study and those of recent independent reports on laurel hydraulics

Bone regeneration with adipose derived stem cells in a rabbit model

It has been shown that stem cells are able to calcify both in vitro and in vivo once implanted under the skin, if conveniently differentiated. Nowadays, however, a study on their efficiency in osseous regeneration does not exist in scientific literature and this very task is the real aim of the present experimentation. Five different defects of 6 mm in diameter and 2 mm in depth were created in the calvaria of 8 white New Zealand rabbits. Four defects were regenerated using 2 different conveniently modified scaffolds (Bio-Oss\uae Block and Bio-Oss Collagen\uae, Geistlich), with and without the aid of stem cells. After the insertion, the part was covered with a collagen membrane fixed by 5 modified titan pins (Altapin\uae). The defect in the front was left empty on purpose as an internal control to each animal. Two animals were sacrificed respectively after 2, 4, 6, 10 weeks. The samples were evaluated with micro-CT and histological analysis. Micro-CT analysis revealed that the quantity of new bone for samples with Bio-Oss\uae Block and stem cells was higher than for samples with Bio-Oss\uae Block alone. Histological analysis showed that regeneration occurred in an optimal way in every sample treated with scaffolds. The findings indicated that the use of adult stem cells combined with scaffolds accelerated some steps in normal osseous regeneration