118 resultados para Biphasic scaffold


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Chikungunya virus (CHIKV) is a mosquito-borne arthrogenic alphavirus that causes acute febrile illness in humans accompanied by joint pains and in many cases, persistent arthralgia lasting weeks to years. The re-emergence of CHIKV has resulted in numerous outbreaks in the eastern hemisphere, and threatens to expand in the foreseeable future. Unfortunately, no effective treatment is currently available. The present study reports the use of resazurin in a cell-based high-throughput assay, and an image-based high-content assay to identify and characterize inhibitors of CHIKV-infection in vitro. CHIKV is a highly cytopathic virus that rapidly kills infected cells. Thus, cell viability of HuH-7 cells infected with CHIKV in the presence of compounds was determined by measuring metabolic reduction of resazurin to identify inhibitors of CHIKV-associated cell death. A kinase inhibitor library of 4,000 compounds was screened against CHIKV infection of HuH-7 cells using the resazurin reduction assay, and the cell toxicity was also measured in non-infected cells. Seventy-two compounds showing >= 50% inhibition property against CHIKV at 10 mu M were selected as primary hits. Four compounds having a benzofuran core scaffold (CND0335, CND0364, CND0366 and CND0415), one pyrrolopyridine (CND0545) and one thiazol-carboxamide (CND3514) inhibited CHIKV-associated cell death in a dose-dependent manner, with EC50 values between 2.2 mu M and 7.1 mu M. Based on image analysis, these 6 hit compounds did not inhibit CHIKV replication in the host cell. However, CHIKV-infected cells manifested less prominent apoptotic blebs typical of CHIKV cytopathic effect compared with the control infection. Moreover, treatment with these compounds reduced viral titers in the medium of CHIKV-infected cells by up to 100-fold. In conclusion, this cell-based high-throughput screening assay using resazurin, combined with the image-based high content assay approach identified compounds against CHIKV having a novel antiviral activity -inhibition of virus-induced CPE - likely by targeting kinases involved in apoptosis.

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Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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This review is intended to gather together recent studies that explore the effects of different types of noninvasive ventilation (NIV) on the autonomic nervous system, assessed through heart rate variability (HRV). A search for papers was conducted in the PubMed, PEDro, SciELO and Lilacs databases with the following descriptors: noninvasive ventilation, CPAP ventilation, intermittent positive pressure breathing and autonomic nervous system, for the period between 2008 and 2012. After eliminating papers not addressing the topic, we selected six studies, of which five applied NIV in CPAP mode and one used biphasic positive airway pressure. In general, the findings suggest that NIV promotes changes in autonomic modulation that are dependent on the conditions of the subjects analyzed and the time when these rates are assessed, meaning acute or long-term effects.

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Stemming from in vitro and in vivo pre-clinical and human models, tissue-engineering-based strategies continue to demonstrate great potential for the regeneration of the pulp-dentin complex, particularly in necrotic, immature permanent teeth. Nanofibrous scaffolds, which closely resemble the native extracellular matrix, have been successfully synthesized by various techniques, including but not limited to electrospinning. A common goal in scaffold synthesis has been the notion of promoting cell guidance through the careful design and use of a collection of biochemical and physical cues capable of governing and stimulating specific events at the cellular and tissue levels. The latest advances in processing technologies allow for the fabrication of scaffolds where selected bioactive molecules can be delivered locally, thus increasing the possibilities for clinical success. Though electrospun scaffolds have not yet been tested in vivo in either human or animal pulpless models in immature permanent teeth, recent studies have highlighted their regenerative potential both from an in vitro and in vivo (i.e., subcutaneous model) standpoint. Possible applications for these bioactive scaffolds continue to evolve, with significant prospects related to the regeneration of both dentin and pulp tissue and, more recently, to root canal disinfection. Nonetheless, no single implantable scaffold can consistently guide the coordinated growth and development of the multiple tissue types involved in the functional regeneration of the pulp-dentin complex. The purpose of this review is to provide a comprehensive perspective on the latest discoveries related to the use of scaffolds and/or stem cells in regenerative endodontics. The authors focused this review on bioactive nanofibrous scaffolds, injectable scaffolds and stem cells, and pre-clinical findings using stem-cell-based strategies. These topics are discussed in detail in an attempt to provide future direction and to shed light on their potential translation to clinical settings.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Malformations and possible damages to the urogenital system can be originated in the embryonic period. Moreover, fire guns, knives and accidents, where there is the disruption of the urethra, also cause these lesions. The objective was to analyze the contribution of tissue engineering in the construction of neo-urethra, developed by bioengineering. We performed an urothelial ex vivo expansion of cells in 3D scaffolds (platelet gel matrix and acellular porcine aorta) to assess the contribution of this technique in the construction of a neo-urethra. Mechanical dissociation was made of the inner wall of 10 North Folk rabbit’s bladder, weighing 2.5 to 3.0 kg. After dissociation the cell content was centrifuged and obtained a pellet of urothelial cells. The pellet was ressuspended in culture medium DMEM F12 and cells were maintained in culture for 15 days. Immunohistochemical analysis characterized the urothelial culture. The cells were then implanted in the scaffold - platelet gel. In a second experiment using aortic porcine acellular matrix were implanted urothelial cells alone and urothelial cells on platelet gel, on the inner wall of the scaffold - aorta, with space for setting bordered by a urethral probe. The complex probe - cells - aorta and probe - cells in platelet gel - aorta, were sealed with suture material and culture were maintained in a humidified 37ºC incubator with 5% CO2 in air for 12 days to subsequent histological analysis of urothelium cell adhesion to the scaffolds. By observation under an optical microscope, we could see the growth of cells in the scaffold platelet gel, from a monolayer in to a three-dimensional structure. In the acellular porcine aortic matrix containing the platelet gel, we could observe a few quantity of urothelial cells adhered. However with the acellular porcine aortic matrix in which was implanted only the urothelial cells, we have obtained adhesion to the wall

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Articular cartilage is the structure that coats the bone ends in regions where two bones are articulated, allowing movement. It has inefficient intrinsic and extrinsic mechanisms of repair, usually resulting in fibrocartilage formation after injury. Such repair have lower strength, stiffness and usability features when compared to hyaline cartilage. The mesenchymal stem cells have the potential to regenerate tissue without the production of scar, and because of this feature it is well studied. But to have its maximum chondrogenic potential, it is necessary to use scaffolds and growth factors. Biomaterials play the role of scaffold for the cells allowing them to become attached, grow and produce extracellular matrix, leading to formation of repair with hyaline cartilage. In this sense, the purpose of this study is to provide information on the various studies using cell therapy and / or biomaterials to produce hyaline cartilage

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Liquid-liquid extraction is utilized for purification of biomolecules by Aqueous Biphasic Systems (ABS), since this process does not damage the biotechnological potential of these compounds. In this work, using the free software Scilab®, the fractionated liquid-liquid extraction was studied aiming a lipase (target enzyme) partition in a water + PEG + DEX system. Lisozime was considered as contaminant. As computer simulations has been extensively used when a first estimation of technical feasibility of process is desired, this work demonstred that the system is viable for recovery the 80 % till 90 % the target enzyme, but should be noted that the trial aimed only the fractionation of the enzyme target of a contaminant, not its concentration, which should be done by another process.

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This project developed microstructural characterization technics of commercial dualphase and multi-phasic (TRIP) steels that were provided by the automotive industry and are currently used as the raw material for the production of automobiles. Inserted in this context, there is the development of the advanced high strength steels in consonance with the ULSABAVC project, which aims the production of safe, economically viable and efficient in terms of fuel consumption vehicles for the 21st century. The micro-structural characterization of biphasic and multiphase steels was done by the identification and quantification of the coexistent phases. In this item, a special attention was given to the technics that were performed using optic microscopy and scanning electron microscopy. An important contribution to this work was the utilization of different alternative chemical reagents (Beraha, Heat-Tinting technics) in addition to the classical ones (Nital and LePera)already used conventionally by the UNESP's Group of Mechanical, Microstructural and Fractografic Characterization of Materials. The revealed microstructures were correlated with the materials' mechanical properties determined through traction tests, such as ultimate tensile strengths, yield strength and stretching important since the material has structural application in the automotive industry. As a result, it was observed the superiority concerning the studied mechanical properties for the biphasic and multiphasic steels when compared to the conventional carbon steels. Besides, it was perceived a large potential for the industrial scale utilization of the Heat Tinting technics in this field, seen its differentiation of the existent phases and easy reproducibility

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Pós-graduação em Química - IQ

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The purpose of this literature review is to present the state-of-the-art on the application of tissue engineering techniques in sinus lifts procedures, reporting the influence of these techniques in increasing bone height and volume in the maxillary sinus, and the osseointegration of the implants installed in the grafted areas. PubMed and Scopus databases were searched using the keywords “scaffold” OR “engineered tissue” AND “sinus augmention” OR “sinus floor elevation”. In the initial search 463 articles were selected, of which 19 were selected abstracts reading, with nine papers selected by the end, for evaluation of the application of tissue engineering techniques in sinus lift procedures. Despite the positive results of tissue engineering procedures in sinus floor elevation reported in the selected articles, further studies are still needed, for a better standardization of experimental models and materials used, leading to definitive conclusions about the effects of the application of tissue engineering procedures on bone formation in maxillary sinus lifting procedures.

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The association between tridimensional scaffolds to cells of interest has provided excellent perspectives for obtaining viable complex tissues in vitro, such as skin, resulting in impressive advances in the field of tissue engineering applied to regenerative therapies. The use of multipotent mesenchymal stromal cells in the treatment of dermo-epidermal wounds is particularly promising due to several relevant properties of these cells, such as high capacity of proliferation in culture, potential of differentiation in multiple skin cell types, important paracrine and immunomodulatory effects, among others. Membranes of chitosan complexed with xanthan may be potentially useful as scaffolds for multipotent mesenchymal stromal cells, given that they present suitable physico-chemical characteristics and have adequate tridimensional structure for the adhesion, growth, and maintenance of cell function. Therefore, the purpose of this work was to assess the applicability of bioactive dressings associating dense and porous chitosan-xanthan membranes to multipotent mesenchymal stromal cells for the treatment of skin wounds. The membranes showed to be non-mutagenic and allowed efficient adhesion and proliferation of the mesenchymal stromal cells in vitro. In vivo assays performed with mesenchymal stromal cells grown on the surface of the dense membranes showed acceleration of wound healing in Wistar rats, thus indicating that the use of this cell-scaffold association for tissue engineering purposes is feasible and attractive.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Introduction: Antibiotic-containing polymer-based nanofibers (hereafter referred to as scaffolds) have demonstrated great potential for their use in regenerative endodontics from both an antimicrobial and cytocompatibility perspective. This study sought to evaluate in vitro the effects of ciprofloxacin (CIP)-containing polymer scaffolds against Enterococcus faecalis biofilms. Methods: Human mandibular incisors were longitudinally sectioned to prepare radicular dentin specimens. Sterile dentin specimens were distributed in 24-well plates and inoculated with E. faecalisfor biofilm formation. Infected dentin specimens were exposed to 3 groups of scaffolds, namely polydioxanone (PDS) (control), PDS + 5 wt% CIP, and PDS + 25 wt% CIP for 2 days. Colony-forming units (CFU/mL) (n = 10) and scanning electron microscopy (SEM) (n -= 2) were performed to quantitatively and qualitatively assess the antimicrobial effectiveness, respectively. Results: PDS scaffold containing CIP at 25 wt% showed maximum bacteria elimination with no microbial growth, differing statistically (P < .05) from the control (PDS) and from PDS scaffold containing CIP at 5 wt%. Statistical differences (P < .05) were also seen for the CFU/mL data between pure PDS (5.92-6.02 log CFU/mL) and the PDS scaffold containing CIP at 5 wt% (5.39 5.87 log CFU/mL). SEM images revealed a greater concentration of bacteria on the middle third of the dentin specimen. after 5 days of biofilm formation. On scaffold exposures, SEM images showed similar results when compared with the CFU/mL data. Dentin specimens exposed to PDS + 25 wt% CIP scaffolds displayed a practically bacteria-free surface. Conclusions: On the basis of the data presented, newly developed antibiotic-containing electrospun scaffolds hold promise as an intracanal medicament to eliminate biofilm/infection before regenerative procedures.