Genetic characterization of alpine sheep breeds

The aim of this study was to characterize from the genetic point of view eight alpine sheep breeds reared in Italy (Bergamasca, Biellese, Schwarzbraunes Bergschaf and Tiroler Bergschaf), Germany (Brillenschaf and Weisses Bergschaf) and Slovenia (Bov\u161ka and Jezerzko-Sol\u10davska) through the use of microsatellite molecular markers. Allelic richness was rather high in each breed highlighting a considerable genetic diversity. However, the study evidenced a significant departure from Hardy-Weinberg equilibrium in all analyzed breeds caused by a heterozygote deficiency. Such lack seems to be caused by a rather high level of inbreeding. The genetic differentiation among breeds was rather low (F ST = 0.064) but significant. The neighbour-joining tree obtained from Reynolds\u2019 genetic distance estimates, showed the presence of three groups formed by the three Bergschaf breeds, the Italian Bergamasca and Biellese and the two Slovenian breeds together with the German Brillenschaf. Such grouping is in accordance with the breeds\u2019 region of origin and with their known history. Concluding, microsatellite resulted to be a useful tool to investigate breed variability and to characterize alpine sheep breeds. Obtained findings suggest the need to set up a conservation plan aiming to safeguard and increase the genetic variability of the studied breeds compromised by the high level of inbreeding. Microsatellites genotyping could help to monitor breed variability and to organize matings

3D collagen cultures under well-defined dynamic strain: A novel strain device with a porous elastomeric support

The field of mechanobiology has grown tremendously in the past few decades, and it is now well accepted that dynamic stresses and strains can impact cell and tissue organization, cell-cell and cell-matrix communication, matrix remodeling, cell proliferation and apoptosis, cell migration, and many other cell behaviors in both physiological and pathophysiological situations. Natural reconstituted matrices like collagen and fibrin are often used for three-dimensional (3D) mechanobiology studies because they naturally form fibrous architectures and are rich in cell adhesion sites; however, they are physically weak and typically contain >99% water, making it difficult to apply dynamic stresses to them in a truly 3D context. Here we present a composite matrix and strain device that can support natural matrices within a macroporous elastic structure of polyurethane. We characterize this system both in terms of its mechanical behavior and its ability to support the growth and in vivo-like behaviors of primary human lung fibroblasts cultured in collagen. The porous polyurethane was created with highly interconnected pores in the hundreds of mm size scale, so that while it did not affect cell behavior in the collagen gel within the pores, it could control the overall elastic behavior of the entire tissue culture system. In this way, a well-defined dynamic strain could be imposed on the 3D collagen and cells within the collagen for several days (with elastic recoil driven by the polyurethane) without the typical matrix contraction by fibroblasts when cultured in 3D collagen gels. We show lung fibroblast-to-myofibroblast differentiation under 30%, 0.1 Hz dynamic strain to validate the model and demonstrate its usefulness for a wide range of tissue engineering applications. © 2008 Wiley Periodicals, Inc

Chitosan-hydroxyapatite composites made from sustainable sources: a morphology and antibacterial study

Chitosan (Cs) and hydroxyapatite (HA) 3D scaffolds/composites were prepared with a sustainable process, as HA was obtained using CaCO3 derived from cork, a natural material used as a template agent. The HA@Cs composites were prepared with HA in situ formation in a Cs solution, with a dissolution-precipitation mechanism. Different reaction times were considered, with time of 72 h leading to the best materials (sample CsHA_72). X-ray Diffraction (XRD) analysis confirmed HA formation. The analysis of Cs unit cell parameters showed that, for the unmodified Cs, the cell had larger dimensions and a higher degree of distortion than previously reported in literature; HA incorporation in the CsHA_72 composite led to a further increase in the cell dimensions. The morphology of the scaffolds was studied with Scanning Electron Microscopy (SEM) and a high level of porosity was observed; a statistical comparison was performed between the unmodified Cs and CsHA_72 to determine the pore size, structure, and distribution. This analysis, the first of this kind for this type of composites, showed smaller and more circular pores for the CsHA_72 composite (average diameter of 70 μm vs. 88 μm for unmodified Cs). The overall level of porosity, however, did not change (>77%); likewise, the Young modulus was not affected by HA incorporation (about 11 kPa). Antibacterial tests, performed on Escherichia coli and Staphylococcus aureus, showed that HA presence did not significantly reduce the antimicrobial properties; the composites were particularly effective towards S. aureus, as a >90% the bacterial population reduction was observed for an incubation time of 2 h. HA@Cs also showed excellent biocompatibility and good cell proliferation. The properties of these 3D scaffolds make them suitable for application as biomaterials.info:eu-repo/semantics/publishedVersio

Post-progression evaluation of patients treated with nivolumab for advanced non-small cell lung cancer: A prospective cohort analysis

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Evaluation of in vivo response of three biphasic scaffolds for osteochondral tissue regeneration in a sheep model

Osteochondral defects are a common problem in both human medicine and veterinary practice although with important limits concerning the cartilaginous tissue regeneration. Interest in the subchondral bone has grown, as it is now considered a key element in the osteochondral defect healing. The aim of this work was to generate and to evaluate the architecture of three cell-free scaffolds made of collagen, magnesium/hydroxyapatite and collagen hydroxyapatite/wollastonite to be implanted in a sheep animal model. Scaffolds were designed in a bilayer configuration and a novel "Honey" configuration, where columns of hydroxyapatite were inserted within the collagen matrix. The use of different types of scaffolds allowed us to identify the best scaffold in terms of integration and tissue regeneration. The animals included were divided into four groups: three were treated using different types of scaffold while one was left untreated and represented the control group. Evaluations were made at 3 months through CT analysis. The novel "Honey" configuration of the scaffold with hydroxyapatite seems to allow for a better reparative process, although we are still far from obtaining a complete restoration of the defect at this time point of follow-up

A tissue engineered osteochondral composite for cartilage repair: an in vivo study

This work aimed to validate the efficacy of a tissue engineered osteochondral composite for the treatment of cartilage lesion produced in adult pigs. The osteochondral composite was manufactured by combining an osteo-compatible cylinder and a neocartilagineous tissue obtained by seeding swine articular chondrocytes into a collagen scaffold. Articular cartilage was harvested from the trochlea of six adult pigs and was enzymatically digested to isolate the chondrocytes [Deponti D.et al. 2005]. The cells were then expanded in monolayer culture in chondrogenic medium and seeded onto a collagen scaffold. The collagen scaffold was preintegrated in vitro, macroscopically and microscopically, to a an osteo-compatible cylinder. The seeded osteochondral scaffolds were left in standard culture condition for 3 weeks with the addition of growth factors. At the end of culture time the osteochondral scaffolds were surgically implanted in osteochondral lesion performed in the trochlea of the same pigs from which the cartilage was initially harvested. As control, some osteochondral lesions were treated with acellular scaffolds and others were left untreated. After 3 months, the repair tissue of the three experimental groups was macroscopically analyzed and processed for histological and biochemical analysis. The hystologic ICRS II scale showed a statistically significant difference between the three experimental groups only in the parameters regarding the cell morphology and the surface/superficial assessment: the lesion treated with the unseeded osteochondral scaffolds showed higher values in chondrocytes morphology and in the superficial layer recovery, with respect to the lesions treated with the seeded scaffolds or left untreated. The biochemical analysis showed a higher DNA content in the lesion repaired with cellular scaffold and a higher GAGs/DNA ratio in the lesions with a spontaneous repair. The result of this study demonstrate that an osteochondral scaffold was able to repair an osteochondral lesion in an in vivo model of adult pigs, showing a good integration with the surrounding tissue. The quality of the repair was higher when the scaffold was not seeded with chondrocytes, but filled with cells migrated from subchondral bone. This tissue engineered osteochondral composite could represent a valuable model for further in vivo studies on the repair of chondral/osteochondral lesion

Mechanical, pH and Thermal Stability of Mesoporous Hydroxyapatite

The stability of mesoporous hydroxyapatite (HAP) powder was studied following treatments of ultrasound, pH and heating. HAP was found to be mechanically stable up to (and including) 1 h continuous ultrasonic treatment in water. The HAP structure was also stable to pH, evidenced by practically identical XRD and FTIR spectra over the pH range 2–12. The surface area increased progressively with increasing acidity, reaching a maximum of 121.9 m 2 g −1 at pH 2, while alkaline conditions decreased the surface area to a minimum of 55.4 m 2 g −1 at pH 12. Heating in air had a significant influence on the structural and morphological properties of HAP, which underwent dehydroxylation to form oxyhydroxyapatite (OHAP) at temperatures ≥ 650 °C, and β-tricalcium phosphate (β-TCP) ≥750 °C. The surface area decreased at elevated temperatures due to agglomeration of HAP crystals by sintering, which was associated with an increased particle size

Sequential Structural and Fluid Dynamics Analysis of Balloon-Expandable Coronary Stents: A Multivariable Statistical Analysis

Several clinical studies have identified a strong correlation between neointimal hyperplasia following coronary stent deployment and both stent-induced arterial injury and altered vessel hemodynamics. As such, the sequential structural and fluid dynamics analysis of balloon-expandable stent deployment should provide a comprehensive indication of stent performance. Despite this observation, very few numerical studies of balloon-expandable coronary stents have considered both the mechanical and hemodynamic impact of stent deployment. Furthermore, in the few studies that have considered both phenomena, only a small number of stents have been considered. In this study, a sequential structural and fluid dynamics analysis methodology was employed to compare both the mechanical and hemodynamic impact of six balloon-expandable coronary stents. To investigate the relationship between stent design and performance, several common stent design properties were then identified and the dependence between these properties and both the mechanical and hemodynamic variables of interest was evaluated using statistical measures of correlation. Following the completion of the numerical analyses, stent strut thickness was identified as the only common design property that demonstrated a strong dependence with either the mean equivalent stress predicted in the artery wall or the mean relative residence time predicted on the luminal surface of the artery. These results corroborate the findings of the large-scale ISAR-STEREO clinical studies and highlight the crucial role of strut thickness in coronary stent design. The sequential structural and fluid dynamics analysis methodology and the multivariable statistical treatment of the results described in this study should prove useful in the design of future balloon-expandable coronary stents

The biomaterialist’s task: scaffold biomaterials and fabrication technologies

This paper focuses on tissue engineering (TE) from the biomaterialist’s point of view. With the aim of answering some simple but key questions about TE, the related literature is here reviewed. In order to obtain an engineered tissue the following steps are mandatory: (a) cell selection, (b) identification of the ideal three-dimensional scaffold for cell seeding and proliferation, (c) choice of the most suitable type of cell culture. Whereas the biotechnologist working in the TE field is responsible for optimizing the cell seeding and culture, the biomaterialist has the challenging task of optimizing the three-dimensional cell support, or scaffold. Therefore, in the present paper, scaffold properties, biomaterials and fabrication technologies are analyzed in depth and reviewed on the basis of the current literature. Finally, mention is also made of the most recently emerging and innovative technologies relating to scaffolds for TE applications