Rapid production of human liver scaffolds for functional tissue engineering by high shear stress oscillation-decellularization

The development of human liver scaffolds retaining their 3-dimensional structure and extra-cellular matrix (ECM) composition is essential for the advancement of liver tissue engineering. We report the design and validation of a new methodology for the rapid and accurate production of human acellular liver tissue cubes (ALTCs) using normal liver tissue unsuitable for transplantation. The application of high shear stress is a key methodological determinant accelerating the process of tissue decellularization while maintaining ECM protein composition, 3D-architecture and physico-chemical properties of the native tissue. ALTCs were engineered with human parenchymal and non-parenchymal liver cell lines (HepG2 and LX2 cells, respectively), human umbilical vein endothelial cells (HUVEC), as well as primary human hepatocytes and hepatic stellate cells. Both parenchymal and non-parenchymal liver cells grown in ALTCs exhibited markedly different gene expression when compared to standard 2D cell cultures. Remarkably, HUVEC cells naturally migrated in the ECM scaffold and spontaneously repopulated the lining of decellularized vessels. The metabolic function and protein synthesis of engineered liver scaffolds with human primary hepatocytes reseeded under dynamic conditions were maintained. These results provide a solid basis for the establishment of effective protocols aimed at recreating human liver tissue in vitro

Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein

A method for non-invasive visualization of genetically labelled cells in animal disease models with micron-level resolution would greatly facilitate development of cell-based therapies. Imaging of fluorescent proteins (FPs) using red excitation light in the “optical window” above 600 nm is one potential method for visualizing implanted cells. However, previous efforts to engineer FPs with peak excitation beyond 600 nm have resulted in undesirable reductions in brightness. Here we report three new red-excitable monomeric FPs obtained by structure-guided mutagenesis of mNeptune, previously the brightest monomeric FP when excited beyond 600 nm. Two of these, mNeptune2 and mNeptune2.5, demonstrate improved maturation and brighter fluorescence, while the third, mCardinal, has a red-shifted excitation spectrum without reduction in brightness. We show that mCardinal can be used to non-invasively and longitudinally visualize the differentiation of myoblasts and stem cells into myocytes in living mice with high anatomical detail

Estudo comparativo entre a agressividade superficial obtida na retificação com rebolos de óxido de alumínio e CBN, fabricados com ligantes resinóide e vitrificado

Este artigo apresenta um estudo da agressividade do rebolo (habilidade de corte). A retificação é um processo de usinagem preciso, o qual é amplamente usado na manufatura de componentes que requerem tolerâncias estreitas e superfícies bem acabadas. Na retificação, a ferramenta abrasiva é o rebolo, que é basicamente composto por núcleo, ligante e grãos abrasivos. As ferramentas testadas nessa pesquisa foram rebolos convencional (Al2O3) e superabrasivo (CBN). Entre os superabrasivos, três tipos específicos de ligantes foram testados: resinóide, vitrificado e resinóide de alto desempenho. Conseqüentemente, foi possível avaliar a habilidade de corte entre os diferentes tipos de rebolos testados. Assim, o rebolo convencional de óxido de alumínio apresentou a maior agressividade.This paper presents a study of the grinding wheel sharpness (cutting ability). Grinding is a precision machining process which is widely used in the manufacture of components requiring fine tolerances and smooth surfaces. In grinding, the abrasive tool is the grinding wheel, which is basically compounded by the core, the bond and the abrasive grains. The tools tested in this research were conventional (Al2O3) and superabrasive (CBN) grinding wheels. Among the superabrasive ones, three specific bond types were tested: resin, vitrified and high performance resin bond. Consequently, it was possible to evaluate the comparative cutting ability among the different types of grinding wheels tested. So the conventional wheel (Al2O3) presented the highest sharpness.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Analysis of the different forms of application and types of cutting fluid used in plunge cylindrical grinding using conventional and superabrasive CBN grinding wheels

The work reported here involved an investigation into the grinding process, one of the last finishing processes carried out on a production line. Although several input parameters are involved in this process, attention today focuses strongly on the form and amount of cutting fluid employed, since these substances may be seriously pernicious to human health and to the environment, and involve high purchasing and maintenance costs when utilized and stored incorrectly. The type and amount of cutting fluid used directly affect some of the main output variables of the grinding process which are analyzed here, such as tangential cutting force, specific grinding energy, acoustic emission, diametrical wear, roughness, residual stress and scanning electron microscopy. To analyze the influence of these variables, an optimised fluid application methodology was developed (involving rounded 5, 4 and 3 turn diameter nozzles and high fluid application pressures) to reduce the amount of fluid used in the grinding process and improve its performance in comparison with the conventional fluid application method (of diffuser nozzles and lower fluid application pressure). To this end, two types of cutting fluid (a 5% synthetic emulsion and neat oil) and two abrasive tools (an aluminium oxide and a superabrasive CBN grinding wheel) were used. The results revealed that, in every situation, the optimised application of cutting fluid significantly improved the efficiency of the process, particularly the combined use of neat oil and CBN grinding wheel. (c) 2005 Elsevier Ltd. All rights reserved

Rectificacion de aceros fragiles y ductiles: Parametros de entrada para obtener los mejores resultados

In this work, the behavior of the steel surface grinding process with conventional grinding wheels was studied for various grinding conditions and a fixed dressing condition. The input parameters (wheel speed, workpiece speed, depth of cut), that set up the grinding conditions were determined as a function of the characteristics of the grinding machine. A test bench was established where the grinding power was measured in real time as a function of the volume of material removed. The results showed that in the grinding of fragile steels with a soft wheel and medium grit size, the average power of the cut shows a tendency to increase and become stabilized. The surface roughness of the piece decreased in all the tests and was proportional to the increase in the number of active grits. In tests on the grinding of ductile steels with a soft wheel and fine grit size, the wheel became dull until the moment that the abrasive grits were released from the cut surface

The effect of amidic moieties on the polysaccharides: evaluation of the physico-chemical and biological properties of amidic carboxymethylcellulose (CMCA) in the form of linear polymer and hydrogel

A new derivative of carboxymethylcellulose was synthesised by converting some of the carboxylic groups into amidic moieties (CMCA). The synthetic procedure allowed for a good control of the degree of substitution. The new polysaccharide was characterised in terms of physico-chemical properties and biocompatibility. It was also used for the realisation of a hydrogel, and its physicochemical behaviour, such as water uptake and rheological parameters, were evaluated. The new CMCA hydrogel showed physico-chemical properties closer to those of Hyaluronan (Hyal) rather than native carboxymethylcellulose (CMC). Hydrogel cytotoxicity and biological activity were then comparatively investigated on osteoblast-like cells, taking into consideration the possible application of the hydrogel as a cell scaffold in orthopaedic tissue engineering. Finally, the effectiveness of cellular scaffolds for b-pancreatic islet cells was also studied