345 research outputs found

    Neutrino Physics

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    This is the writeup of the lectures on neutrino physics delivered at various schools: TASI and Trieste in 2013 and the CERN-Latin American School in 2015. The topics discussed in this lecture include: general properties of neutrinos in the SM, the theory of neutrino masses and mixings (Dirac and Majorana), neutrino oscillations both in vacuum and in matter, as well as an overview of the experimental evidence for neutrino masses and of the prospects in neutrino oscillation physics. We also briefly review the relevance of neutrinos in leptogenesis and in beyond-the-Standard-Model physics.Comment: 58 pages, contribution to the CERN in the Proceedings of the 2015 CERN-Latin-American School of High-Energy Physics, Ibarra, Ecuador, 4 - 17 March 2015. arXiv admin note: text overlap with arXiv:1010.413

    On the structure of natural human movement

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    Understanding of human motor control is central to neuroscience with strong implications in the fields of medicine, robotics and evolution. It is thus surprising that the vast majority of motor control studies have focussed on human movement in the laboratory while neglecting behaviour in natural environments. We developed an experimental paradigm to quantify human behaviour in high resolution over extended periods of time in ecologically relevant environments. This allows us to discover novel insights and contradictory evidence to well-established findings obtained in controlled laboratory conditions. Using our data, we map the statistics of natural human movement and their variability between people. The variability and complexity of the data recorded in these settings required us to develop new tools to extract meaningful information in an objective, data-driven fashion. Moving from descriptive statistics to structure, we identify stable structures of movement coordination, particularly within the arm-hand area. Combining our data with numerous published findings, we argue that current hypotheses that the brain simplifies motor control problems by dimensionality reduction are too reductionist. We propose an alternative hypothesis derived from sparse coding theory, a concept which has been successfully applied to the sensory system. To investigate this idea, we develop an algorithm for unsupervised identification of sparse structures in natural movement data. Our method outperforms state-of-the-art algorithms for accuracy and data-efficiency. Applying this method to hand data reveals a dictionary of \emph{sparse eigenmotions} (SEMs) which are well preserved across multiple subjects. These are highly efficient and invariant representation of natural movement, and suggest a potential higher-order grammatical structure or ``movement language''. Our findings make a number of testable predictions about neural coding of movement in the cortex. This has direct consequences for advancing research on dextrous prosthetics and robotics, and has profound implications for our understanding of how the brain controls our body.Open Acces

    Shape Memory Polymers as 2D Substrates and 3D Scaffolds for the Study of Cell Mechanobiology and Tissue Engineering

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    Tissue engineering is a promising, fast-growing field that combines cells, signals, and scaffolds to regenerate damaged tissues. To develop new, functional, engineered tissues, it is becoming increasingly important to understand how cell-material interactions affect the cell mechanobiological response. As a result, recent efforts have focused on developing complex synthetic materials that can mimic the dynamic in vivo cell environment. In this work, shape memory polymers (SMPs) were employed to develop dynamic 2D substrates and 3D scaffolds that undergo programmed changes in shape under cell compatible conditions. These substrates and scaffolds were applied in vitro and in vivo to demonstrate their potential as platforms to study cell mechanobiology and as functional tissue engineered constructs. The first part of this dissertation describes the fabrication and application of an SMP bilayer system capable of forming nano-scale wrinkles under cytocompatible conditions. Wrinkled substrates with easily tunable characteristics were employed to control the degree of cell alignment, with increased wrinkle amplitude and wrinkle orientation resulting in increased cell alignment until reaching a point of saturation. Active wrinkling with attached and viable cells was found to enable cell alignment to be “turned-on” on command. Additionally, cell migration on wrinkled substrates was assessed using quantitative, statistical-physics-based metrics which revealed cell motility atop anisotropic wrinkled substrates and which was more oriented and persistent than cell motility atop flat isotropic controls The second part of this dissertation describes the fabrication and application of porous 3D SMPs capable of expanding under physiological temperatures. A modified porogen-leaching approached was employed to fabricate highly porous, interconnected SMP scaffolds with tunable properties. The potential of SMP foams for use as synthetic bone substitutes was demonstrated in a mouse segmental defect model, where expanding foams were deployed intraoperatively to fill and conform to a critical size defect. Stiff SMP foams were able to maintain defect stability in a load-bearing application and integrated with the native bone after 12 weeks. Furthermore, deployable SMP foams showed potential for use as deployable cell-based therapies to facilitate bone repair, as expanding foams were able to support osteogenic differentiation of attached stem cells. This work demonstrates the potential of SMPs to be employed as dynamic materials to study cell-material interactions in dynamic environments and to aid in the development of functional tissue engineered constructs

    Image processing in medicine advances for phenotype characterization, computer-assisted diagnosis and surgical planning

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    En esta Tesis presentamos nuestras contribuciones al estado del arte en procesamiento digital de imágenes médicas, articulando nuestra exposición en torno a los tres principales objetivos de la adquisición de imágenes en medicina: la prevención, el diagnóstico y el tratamiento de las enfermedades. La prevención de la enfermedad se puede conseguir a veces mediante una caracterización cuidadosa de los fenotipos propios de la misma. Tal caracterización a menudo se alcanza a partir de imágenes. Presentamos nuestro trabajo en caracterización del enfisema pulmonar a partir de imágenes TAC (Tomografía Axial Computerizada) de tórax en alta resolución, a través del análisis de las texturas locales de la imagen. Nos proponemos llenar el vacío existente entre la práctica clínica actual, y las sofisticadas pero costosas técnicas de caracterización de regiones texturadas, disponibles en la literatura. Lo hacemos utilizando la distribución local de intensidades como un descriptor adecuado para determinar el grado de destrucción de tejido en pulmones enfisematosos. Se presentan interesantes resultados derivados del análisis de varios cientos de imágenes para niveles variables de severidad de la enfermedad, sugiriendo tanto la validez de nuestras hipótesis, como la pertinencia de este tipo de análisis para la comprensión de la enfermedad pulmonar obstructiva crónica. El procesado de imágenes médicas también puede asistir en el diagnóstico y detección de enfermedades. Presentamos nuestras contribuciones a este campo, que consisten en técnicas de segmentación y cuantificación de imágenes dermatoscópicas de lesiones de la piel. La segmentación se obtiene mediante un novedoso algoritmo basado en contornos activos que explota al máximo el contenido cromático de las imágenes, gracias a la maximización de la discrepancia mediante comparaciones cross-bin. La cuantificación de texturas en lesiones melanocíticas se lleva a cabo utilizando un modelado de los patrones de pigmentación basado en campos aleatorios de Markov, en un esfuerzo por adoptar la tendencia emergente en dermatología: la detección de la malignidad mediante el análisis de la irregularidad de la textura. Los resultados para ambas técnicas son validados con un conjunto significativo de imágenes dermatológicas, sugiriendo líneas interesantes para la detección automática del melanoma maligno. Cuando la enfermedad ya está presente, el tratamiento digital de imágenes puede asistir en la planificación quirúrgica y la intervención guiada por imagen. La planificación terapeútica, ejemplicada por la planificación de cirugía plástica usando realidad virtual, se aborda en nuestro trabajo en segmentación de hueso/grasa/músculo en imágenes TAC. Usando un abordaje interactivo e incremental, nuestro sistema permite obtener segmentaciones precisas a partir de unos cuantos clics de ratón para una gran variedad de condiciones de adquisición y frente a anatomícas anormales. Presentamos nuestra metodología, y nuestra validación experimental profusa basada tanto en segmentaciones manuales como en valoraciones subjetivas de los usuarios, e indicamos referencias al lector que detallan los beneficios obtenidos con el uso de la plataforma de planifificación que utiliza nuestro algoritmo. Como conclusión presentamos una disertación final sobre la importancia de nuestros resultados y las líneas probables de trabajo futuro hacía el objetivo último de mejorar el cuidado de la salud mediante técnicas de tratamiento digital de imágenes médicas.In this Thesis we present our contributions to the state-of-the-art in medical image processing, articulating our exposition around the three main roles of medical imaging: disease prevention, diagnosis and treatment. Disease prevention can sometimes be achieved by proper characterization of disease phenotypes. Such characterization is often attained from the standpoint of imaging. We present our work in characterization of emphysema from highresolution computed-tomography images via quanti_cation of local texture. We propose to _ll the gap between current clinical practice and sophisticated texture approaches by the use of local intensity distributions as an adequate descriptor for the degree of tissue destruction in the emphysematous lung. Interesting results are presented from the analysis of several hundred datasets of lung CT for varying disease severity, suggesting both the correctness of our hypotheses and the pertinence of _ne emphysema quanti_cation for understanding of chronic obstructive pulmonary disease. Medical image processing can also assist in the diagnosis and detection of disease. We introduce our contributions to this_eld, consisting of segmentation and quanti_cation techniques in application to dermatoscopy images of skin lesions. Segmentation is achieved via a novel active contour algorithm that fully exploits the color content of the images, via cross-bin histogram dissimilarity maximization. Texture quanti_cation in the context of melanocytic lesions is performed using modelization of the pigmentation patterns via Markov random elds, in an e_ort to embrace the emerging trend in dermatology: malignancy assessment based on texture irregularity analysis. Experimental results for both, the segmentation and quanti_cation proposed techniques, will be validated on a signi_cant set of dermatoscopy images, suggesting interesting pathways towards automatic detection and diagnosis of malignant melanoma. Once disease has occurred, image processing can assist in therapeutical planning and image-guided intervention. Therapeutical planning, exempli_ed by virtual reality surgical planning, is tackled by our work in segmentation of bone/fat/muscle in CT images for plastic surgery planning. Using an interactive, incremental approach, our system is able to provide accurate segmentations based on a couple of mouse-clicks for a wide variety of imaging conditions and abnormal anatomies. We present our methodology, and provide profuse experimental validation based on manual segmentations and subjective assessment, and refer the reader to related work reporting on the clinical bene_ts obtained using the virtual reality platform hosting our algorithm. As a conclusion we present a _nal dissertation on the signi_cance of our results and the probable lines of future work towards fully bene_tting healthcare using medical image processing

    Nanofibrous membranes obtained by electrospinning for bone tissue engineering and wound dressing applications

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    Esta tesis doctoral se ha realizado dentro del marco de un acuerdo de co-tutela entre la Universidad de Zaragoza (Universidad de origen), la Universidad de Calabria (Universidad anfitriona) y la Facultad de Ciencias y Tecnología de la Universidad NOVA de Lisboa (FCT NOVA) (Universidad anfitriona). El trabajo de investigación se ha llevado a cabo dentro del programa de Doctorado en Ingeniería de Membranas Erasmus Mundus (EUDIME), (FPA 2011-0014), financiado por la Unión Europea. La tesis se centró principalmente en el uso de la técnica de electrohilado para producir diferentes tipos de membranas que puedan ser utilizadas en distintas aplicaciones biomédicas. Se sintetizaron y produjeron nanopartículas orgánicas e inorgánicas para ser utilizadas como rellenos o como portadores (sistema de administración de fármacos), así como membranas nanofibrosas electrohiladas. Este trabajo se llevó a cabo en el Instituto de Nanociencia de Aragón (INA), específicamente en el grupo de Nanostructured Films and Particles (NFP) bajo la supervisión de la profesora Silvia Irusta y la Dra. Gracia Mendoza. Una parte importante de la caracterización físico-química se realizó en el INA. En la Universidad de Calabria se trabajó bajo la supervisión de la Dra. Loredana de Bartolo en el Instituto de Tecnología de Membranas (ITM). Allí se utilizaron técnicas específicas tanto para la caracterización como para estudiar diferentes señales biológicas producidas por las membranas sintetizadas, bajo la supervisión. Por otro lado, la movilidad llevada a cabo en la Facultad de Ciencias y Tecnología (FCT NOVA) de la Universidade NOVA (FCT NOVA) bajo la supervisión de la profesora Ana Isabel Aguiar-Ricardo, permitió realizar una caracterización completa de dos membranas asimétricas siguiendo diferentes Normas Internacionales que establecen diferentes ensayos a realizar en apósitos primarios utilizados en heridas. El desarrollo de nuevos scaffolds cargados con proteínas morfogenéticas o antibióticos es de gran interés en el campo de la ingeniería de tejidos óseos. Scaffolds electrohilados con una microporosidad mejorada puede ser beneficioso para mejorar la viabilidad celular debido a que una alta porosidad junto a la presencia de microporos puede proporcionar un entorno tridimensional (3D) que no solamente facilita la siembra y difusión celular sino también proporciona una mejor difusión de los nutrientes y residuos a través del scaffolds. La adición de cerámica de fosfato de calcio ha sido ampliamente investigada para fabricar scaffolds altamente porosos para la ingeniería de tejidos óseos debido a que presentan una composición muy similar al hueso, incluyendo excelentes propiedades de biocompatibilidad, osteoinductivas y osteoconductoras. Partículas cargadas con proteínas morfogenéticas de hueso distribuidas homogéneamente en el scaffolds podrían asegurar una liberación continua del factor de crecimiento proporcionando de esta forma las señales bioquímicas necesarias para la reparación y regeneración ósea. Los scaffolds cargados con antibióticos pueden proporcionar una liberación sostenida del fármaco en el sitio de interés, así como el mantenimiento de propiedades osteogénicas mejoradas para la regeneración exitosa del hueso. Evitando de esta forma que se alcancen niveles de toxicidad o niveles ineficaces en la zona de interés, así como la aparición de efectos secundarios indeseados en los pacientes que provocan un rechazo a los tratamientos prolongados de fármacos por vía sistemática (vía oral e intravenosa). Otra aplicación biomédica interesante de las membranas electrohiladas es la fabricación de apósitos inteligentes eficientes para el tratamiento de heridas. Para lograr una curación rápida de la herida es necesario desarrollar membranas apropiadas con poros interconectados capaces de prevenir la deshidratación rápida y la penetración de bacterias. Para mantener un ambiente húmedo en el lecho de la herida se necesita una alta capacidad de absorción y una adecuada transmisión de vapor de agua. Además, si la membrana electrohilada presenta propiedades bactericidas facilitará el proceso de curación. El objetivo principal de esta tesis fue el desarrollo mediante electrohilado de membranas fibrosas con las características apropiadas para ser utilizadas en la ingeniería de tejidos óseos o como apósito para heridas. En los Capítulos II al V se plantean una serie de objetivos específicos con el fin de cumplir el objetivo principal. Este documento de tesis se dividió en las siguientes secciones: CAPÍTULO I, corresponde a la introducción general donde se describen los conceptos de biomateriales, scaffolds, ingeniería de tejidos y el objetivo principal de los sistemas de liberación de fármacos. Así como, la clasificación de los biomateriales y la ingeniería de tejidos según el origen de los materiales. Además se ponen de manifiesto todos los factores que deben tenerse en cuenta para desarrollar y aplicar adecuadamente los apósitos para heridas. Se mencionaron las diferentes técnicas utilizadas en la literatura haciendo énfasis en el uso de electrohilado y electropulverización para producir scaffolds o membranas para su uso en la ingeniería del tejido óseo y como apósitos para heridas. CAPÍTULO II, se enfoca en el desarrollo y mejora de andamios 3D capaces de promover una eficiente regeneración ósea junto con la liberación de antibióticos dirigidos para prevenir la colonización de bacterias. El objetivo de este trabajo fue sintetizar y caracterizar un sistema de liberación de fármacos que consiste en nanofibras electrohiladas de policaprolactona (PCL) decoradas con partículas de poli (ácido láctico-coglicólico) (PLGA) cargadas con rifampicina (RFP). Este material debe promover la reparación ósea evitando el deterioro del scaffolds provocado por una infección. Se realizó la evaluación in vitro de la capacidad bactericida del material electrohilado sintetizado contra bacterias Gram positivas (Staphylococcus aureus) y Gram negativas (Escherichia coli), así como su citocompatibilidad en cultivos 3D con osteoblastos humanos. Estos resultados se enviaron a la Revista de farmacia “International Journal of Pharmaceuitics” para su publicación en formato de artículo y está bajo revisión. CAPÍTULO III, se describe la síntesis y caracterización de membranas con estructura de núcleo-envoltura de PCL y acetato de polivinilo (PVAc) obtenidas por electrohilado. Las fibras se cargaron con nanopartículas de hidroxiapatita sintética (HAn) para aumentar la bioactividad de los materiales. Los scaffolds desarrollados se trataron con ablación láser para crear características topográficas deseadas a nivel micrométrico con el objetivo de favorecer la adhesión y crecimiento celular. Todas las membranas obtenidas presentaron una estructura de poros tridimensionalmente interconectados y el tratamiento con láser provocó un aumento en la viabilidad y densidad celular. Además, el aumento en la biocompatibilidad de los scaffolds sugiere que los microporos pequeños favorecen la adhesión y proliferación celular. Estos resultados fueron publicados en el artículo titulado “Laser-treated electrospun fibers loaded with nano-hydroxyapatite for bone tissue engineering”. Javier Aragon, Nuria Navascues, Gracia Mendoza, Silvia Irusta. International Journal of Pharmaceutics 525,112–122, 2017. DOI:10.1016/j.ijpharm.2017.04.022. CAPÍTULO IV, se refiere al desarrollo de un scaffold electrohilado compuesto por fibras con estructura de núcleo-cubierta de PCL o PCL/PVAc cargado con HAn sintética. Estas fibras se decoraron con partículas de PLGA cargadas con proteína morfogenética ósea 2 (BMP2) mediante el uso simultaneo de electrohilado coaxial y electropulverización. El objetivo de este trabajo fue evaluar las propiedades estructurales y físico-químicas así como el proceso de biodegradación de los nuevos scaffolds desarrollados y su capacidad para abordar las características arquitectónicas, bioquímicas y funcionales del tejido óseo. Para esto, se probó la bioactividad del scaffold mediante el cultivo de osteoblastos humanos sobre ellos y se monitoreo de la viabilidad celular durante 4 semanas. Se evaluó la actividad osteogénica in vitro de las células sembradas sobre los scaffolds determinando la actividad de la fosfatasa alcalina (ALP) y la expresión de osteocalcina (OCN) y osteopontina (OPN) como proteínas osteogénicas. Estos resultados fueron publicados en el artículo titulado “Polymeric electrospun scaffolds for bone morphogenetic protein 2 delivery in bone tissue engineering”. Javier Aragón, Simona Salerno, Loredana De Bartolo, Silvia Irusta and Gracia Mendoza. Journal of Colloid and Interface Science, 531 (2018) 126–137. DOI:10.1016/j.jcis.2018.07.029. El CAPÍTULO V, describe la síntesis de un apósito antimicrobiano para heridas, con una resistencia mecánica adecuada que es capaz de absorber exudados y evitar la deshidratación rápida de una herida. Se prepararon membranas asimétricas de PCL/PVAc cargadas con carvacrol (CRV) mediante el uso simultáneo de electrohilado y electropulverización. Las membranas constan de dos capas; la primera es una capa de PCL electrohilado; la segunda, una lámina de PVAc que estaría en contacto con la piel liberando a su vez el compuesto antimicrobiano. Se demostró que el uso de diferentes disolventes pueden dar lugar a la obtención de diferentes morfologías de la capa PVAc-CRV. Los valores obtenidos de elongación máxima de las membranas antes de romperse son adecuados para ser utilizados como apósitos para heridas ya que están en el mismo rango reportado de elongaciones en la piel humana. Las membranas presentan una tasa óptima de Transmisión de vapor de agua (WVTR) con valores que se encuentran en el rango requerido para mantener un buen balance entre humedad y pérdida de agua en la herida. En la primera semana, se liberó más del 60 % del CRV cargado, mientras que después de tres semanas, las membranas liberaron entre el 85 y el 100 % del CRV cargado mediante la contribución de un proceso de difusión de tipo Fickiano y la relajación delas cadenas poliméricas. Las membranas sintetizadas son candidatas potenciales para ser utilizadas como apósitos para heridas. El manuscrito que resume estos resultados se envió a la revista “Materials Science and Engineering C” y está bajo revisión (MSEC_2018_3013). CAPÍTULO VI, resume las conclusiones generales del trabajo de tesis. APÉNDICE 1, describe las principales técnicas de caracterización y los métodos para evaluar diferentes propiedades en función de las posibles aplicaciones. APÉNDICE 2, resume los artículos publicados y la participación en foros científicos durante el período de tesis. 1The current Doctoral Thesis work has been performed under a co-supervision agreement between University of Zaragoza (Home University), University of Calabria (Host University) and Faculty of Sciences and Technology of the NOVA University of Lisbon (FCT NOVA) (Host University). This research has been carried out inside the Erasmus Mundus Doctorate in Membrane Engineering program (EUDIME), (FPA 2011-0014), funded by the European Union. This thesis focused mainly on the use of the electrospinning technique to produce different kind of membranes for biomedical applications. In particular, it described the synthesis and production of inorganic and organic nanoparticles to be used as fillers or as carriers (drug delivery system) as well as the production of electrospun nanofibrous membranes. This work was carried out within the Institute of Nanoscience of Aragon (INA), specifically in the Nanostructured Films and Particles (NFP) group under the supervision of the Professor Silvia Irusta and Dr Gracia Mendoza. Also an important part of the physico-chemical characterization was done at INA.The study of different biological signals and the use of specific techniques for membrane characterization were acquired at the University of Calabria under the supervision of Dr. Loredana De Bartolo in the Institute on Membrane Technology of the National Research Council of Italy (ITM-CNR). On the other hand, the mobility carried out at the Faculty of Sciences and Technology (FCT NOVA) of Universidade NOVA (FCT NOVA) under the supervision of Professor Ana Isabel Aguiar-Ricardo, allowed a total characterization of two asymmetric membranes following different International Standards to accomplish testing for primary wound dressing.The development of novel membranes loaded with morphogenetic proteins or antibiotic are of great interest in the field of bone tissue engineering. To promote the cellular viability and extracellular matrix production, electrospun membranes with enhanced porosity and micro-scale pores could be beneficial since increased porosity and pore size can provide a three-dimensional (3D) environment that not only facilitates cell seeding/diffusion but also provides better diffusion of nutrients and waste throughout the membranes. The addition of calcium phosphate ceramics has been extensively investigated to fabricate highly porous membranes to bone tissue engineering due to their close similar composition of bone, including excellent biocompatibility, osteoinductive and osteoconductive properties. A homogeneous distribution of the bone morphogenetic protein-loaded particles along the entire membrane could be ensuring a continuous release of the growth factor to provide the necessary biochemical cues for bone repair and regeneration.Antibiotic-loaded membranes may provide drug targeted and sustained release, avoiding the long-term oral and intravenous systematic multidrug administration, which implies toxic side effects, low delivery to the target site and low patient adherence to the treatment. Therefore, membranes loaded with antibiotics can overcome the drawbacks of the traditional therapy sustaining enhanced osteogenic properties for the successful regeneration of the bone. Another interesting biomedical application of electrospun membranes is the fabrication of efficient smart dressings for the treatment of wounds. A rapid wound healing requires developing appropriate membranes with interconnected pores that allow the oxygen diffusion and transport of metabolic waste, as well as an adequate pore size to prevent rapid dehydration and bacteria penetration. A high absorption capacity and adequate water vapor transmission will be necessary to keep a moist environment in the wound bed. Besides, if the electrospun membrane has some bactericidal properties will be better for the healing process.The main goal of this thesis was the development of fibrous membranes by electrospinning with the appropriate characteristics to be used in bone tissue engineering or as wound dressing materials. To achieve this target, several specific objectives were defined, which are described in Chapters II to V.The thesis was divided in the following sections: CHAPTER I, is an introduction where the concepts of biomaterials, scaffolds and tissue engineering and the main target of drug delivery systems are described. The chapter includes the classification of biomaterials according to the origin of the materials and tissue engineering is also described as well as all the factors that must be taken into account to develop and properly apply a wound dressing are discussed. Different kind of techniques used in the literature to produce scaffolds or membranes for bone tissue engineering and wound dressings are mentioned, focusing on the use of electrospinning and electrospray to produce them. CHAPTER II, focuses on the development of enhanced 3D membranes able to promote efficient bone regeneration together with targeted antibiotic release to prevent bacteria colonization. The aim of this work was to synthesize and characterize a drug delivery system consisting of polycaprolactone (PCL) electrospun nanofibers decorated with rifampicin (RFP) loaded into poly(lactic-coglicolic acid) (PLGA) particles. This material would promote bone repair avoiding the impairment of the membrane mediated by infection. The bactericidal ability of the synthesized electrospun material was assessed In vitro against gram positive (Staphylococcus aureus) and gram negative (Escherichia coli) bacteria, as well as its cytocompatibility in human osteoblasts 3D cultures. These results are included in the accepted article entitled “Composite scaffold obtained by electro-hydrodynamic technique for infection prevention and treatment in bone repair”. Javier Aragon, Sergio Feoli, Gracia Mendoza, Silvia Irusta. International Journal of Pharmaceutics. CHAPTER III, describes the synthesis and characterization of core-shell membranes of PCL and polyvinyl acetate (PVAc) obtained by electrospinning. The fibers were loaded with synthetic hydroxyapatite nanoparticles (HAn) to increase the bioactivity of the materials. The prepared membranes were then treated by laser ablation to create desired microscale topographical features in order to favor cell adhesion and growth. All prepared membranes exhibited a three-dimensional network structure with interconnected pores; the laser treatment has modified the structural characteristics of the membrane causing an increase the cell viability and cell density. The materials biocompatibility is affected by the structural properties of the membranes, indeed smaller micropore sizes favor cell adhesion and proliferation. These results are published in the article entitled “Laser-treated electrospun fibers loaded with nano-hydroxyapatite for bone tissue engineering”. Javier Aragon, Nuria Navascues, Gracia Mendoza, Silvia Irusta. International Journal of Pharmaceutics 525,112–122, 2017. DOI:10.1016/j.ijpharm.2017.04.022. CHAPTER IV, refers to the development of a composite electrospun membrane of PCL or PCL/PVAc core–shell fibers loaded with synthetic HAn. These fibers were decorated with bone morphogenetic protein 2 (BMP2) loaded in/into PLGA particles via simultaneous electrospraying and coaxial electrospinning. The aim of this study was to evaluate the structural and physico-chemical properties and biodegradation processes of the newly developed membranes assessing their ability to address the architectural, biochemical, and functional features of bone tissue. For this purpose, the membrane bioactivity was tested by culturing human osteoblasts on the membranes and by monitoring cell viability up to 4 weeks. The In vitro osteogenic activity of cells seeded onto the membranes was evaluated by assessing alkaline phosphatase (ALP) activity and the expression of osteogenic proteins osteocalcin (OCN) and osteopontin (OPN). These results are published in the article “Polymeric electrospun scaffolds for bone morphogenetic protein 2 delivery in bone tissue engineering”. Javier Aragón, Simona Salerno, Loredana De Bartolo, Silvia Irusta and Gracia Mendoza. Journal of Colloid and Interface Science, 531 (2018) 126–137. DOI:10.1016/j.jcis.2018.07.029. CHAPTER V, describes the synthesis of an antimicrobial wound dressing material, with appropriate mechanical resistance avoiding rapid dehydration and absorbing exudates. PCL/PVAc asymmetric membranes loaded with carvacrol (CRV) were prepared by electrospinning and electrospraying simultaneously. The membranes consist of two layers: the first is an electrospun PCL sheet, the second a PVAc sheet that would be in contact with the skin releasing the antimicrobial compound. The use of different solvents results in different morphologies for the PVAc-CRV layer. The membranes exhibit mechanical properties with strain to failure values that are in the range of human skin, being adequate to be deposited over a wound surface. The samples present Water Vapor Transmission (WVTR) values in the required range to keep good moisture balance with water loss from the wound at the optimal rate. In the first week, more than 60 % of the loaded CRV was released while after three weeks membranes released between 85 to 100 % of the loaded CRV through a Fickian diffusion and diffusion due to polymer relaxation. The synthesized membranes are potential candidates to be used for wound dressing applications. The manuscript summing up these results has been submitted to a scientific journal and is currently under review. GENERAL CONCLUSIONS, summarizes the conclusions of the thesis work. APPENDIX 1, describes the main characterization techniques and the methods to evaluate different properties according to the possible applications. APPENDIX 2, summarizes the articles published and the participation in scientific forums during the thesis period.<br /

    Amélioration de l'image et la segmentation (applications en imagerie médicale)

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    Avancement dans l'acquisition d'image et le progrès dans les méthodes de traitement d'image ont apporté les mathématiciens et les informaticiens dans les domaines qui sont d'une importance énorme pour les médecins et les biologistes. Le diagnostic précoce de maladies (comme la cécité, le cancer et les problèmes digestifs) ont été des domaines d'intérêt en médecine. Développement des équipements comme microscope bi-photonique à balayage laser et microscope de fluorescence par réflexion totale interne fournit déjà une bonne idée des caractéristiques très intéressantes sur l'objet observé. Cependant, certaines images ne sont pas appropriés pour extraire suffisamment d'informations sur de cette image. Les méthodes de traitement d'image ont été fournit un bon soutien à extraire des informations utiles sur les objets d'intérêt dans ces images biologiques. Rapide méthodes de calcul permettent l'analyse complète, dans un temps très court, d'une série d'images, offrant une assez bonne idée sur les caractéristiques souhaitées. La thèse porte sur l'application de ces méthodes dans trois séries d'images destinées à trois différents types de diagnostic ou d'inférence. Tout d'abord, Images de RP-muté rétine ont été traités pour la détection des cônes, où il n'y avait pas de bâtonnets présents. Le logiciel a été capable de détecter et de compter le nombre de cônes dans chaque image. Deuxièmement, un processus de gastrulation chez la drosophile a été étudié pour observer toute la mitose et les résultats étaient cohérents avec les recherches récentes. Enfin, une autre série d'images ont été traités où la source était une vidéo à partir d'un microscopie photonique à balayage laser. Dans cette vidéo, des objets d'intérêt sont des cellules biologiques. L'idée était de suivre les cellules si elles subissent une mitose. La position de la cellule, la dispersion spatiale et parfois le contour de la membrane cellulaire sont globalement les facteurs limitant la précision dans cette vidéo. Des méthodes appropriées d'amélioration de l'image et de segmentation ont été choisies pour développer une méthode de calcul pour observer cette mitose. L'intervention humaine peut être requise pour éliminer toute inférence fausse.Advancement in Image Acquisition Equipment and progress in Image Processing Methods have brought the mathematicians and computer scientists into areas which are of huge importance for physicians and biologists. Early diagnosis of diseases like blindness, cancer and digestive problems have been areas of interest in medicine. Development of Laser Photon Microscopy and other advanced equipment already provides a good idea of very interesting characteristics of the object being viewed. Still certain images are not suitable to extract sufficient information out of that image. Image Processing methods have been providing good support to provide useful information about the objects of interest in these biological images. Fast computational methods allow complete analysis, in a very short time, of a series of images, providing a reasonably good idea about the desired characteristics. The thesis covers application of these methods in 3 series of images intended for 3 different types of diagnosis or inference. Firstly, Images of RP-mutated retina were treated for detection of rods, where there were no cones present. The software was able to detect and count the number of cones in each frame. Secondly, a gastrulation process in drosophila was studied to observe any mitosis and results were consistent with recent research. Finally, another series of images were treated where biological cells were observed to undergo mitosis. The source was a video from a photon laser microscope. In this video, objects of interest were biological cells. The idea was to track the cells if they undergo mitosis. Cell position, spacing and sometimes contour of the cell membrane are broadly the factors limiting the accuracy in this video. Appropriate method of image enhancement and segmentation were chosen to develop a computational method to observe this mitosis. Cases where human intervention may be required have been proposed to eliminate any false inference.SAVOIE-SCD - Bib.électronique (730659901) / SudocGRENOBLE1/INP-Bib.électronique (384210012) / SudocGRENOBLE2/3-Bib.électronique (384219901) / SudocSudocFranceF

    Mechanochemical Control of Stem Cell Biology in Development and Disease: Experimental and Theoretical Models

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    Whether a stem cell remains or egresses away from its physiological niche is a function of mechanical and soluble factors in a time-dependent manner, which implicates a `memory\u27 of prior mechanochemical conditioning. Virtually every organ in the body contains resident stem or progenitor cells that contribute to organ homeostasis or repair. The wound healing process in higher vertebrate animals is spatiotemporally complex and usually leads to scarring. Limitations for the use of stem cells as regenerative therapy include the lack of expansion capabilities in vitro as well as materials issues that complicate traditional biochemical protocols. A minimal `scar in a dish\u27 model is developed to clarify the kinetics of tension-sensitive proteins in mesenchymal stem cells (MSCs), which possess plasticity to mechanochemical changes of the microenvironment that are typical of scars. The organization and expression of such proteins implicates transcription factors that ultimately steer cell fate. In contrast to classic mechano-transducers of matrix mechanics such as actin assembly-dependent serum response factor (SRF) signaling, a novel mechano-repressive role of NKX2.5 is implicated in maintaining intracellular tension in long-term stem cell cultures on stiff matrices via nucleo-cytoplasmic shuttling — ultimately setting up a \u27mechanical memory\u27. Core gene circuits with known roles in stem cell mechanobiology are modeled based on the \u27use it or lose it\u27 concept: tension inhibits turnover of structural proteins such as extracellular collagens, cytoskeletal myosins and nucleoskeletal lamins. This theoretical approach is tested in a variety of processes in vitro and in vivo that involve forces including cardiac development, osteogenic commitment of MSCs, and fibrosis therapy. With the sophistication of the science and technology of biomaterials relevant to stem cell biology and medicine, matrix mechanics can thus be rigorously combined with biochemical instructions in order to maximize therapeutic utility of stem cells

    Framework for neurosphere growth modelling under phase-contrast microscopy

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    L'étude des cellules souches est l'un des champs de recherches les plus importants dans le domaine biomédical. La vision par ordinateur et le traitement d'images ont été fortement mis en avant dans ce domaine pour le développement de solutions automatiques de culture et d'observation de cellules. Ce travail de thèse propose une nouvelle méthodologie pour l'observation et la modélisation de la prolifération de cellule souche neuronale sous microscope à contraste de phase. À chaque observation réalisée par le microscope durant la prolifération, notre système extrait un modèle en trois dimensions de la structure de cellules observées. Cela est réalisé par une suite de processus d'analyse, synthèse et sélection. Premièrement, une analyse de la séquence d'images de contraste de phase permet la segmentation de la neurosphère et des cellules la constituant. À partir de ces informations, combinées avec des connaissances a priori sur les cellules et le protocole de culture, plusieurs modèles 3-D possibles sont générés. Ces modèles sont finalement évalués et sélectionnés par rapport à l¿image d¿observation, grâce à une méthode de recalage 3-D vers 2-D. A travers cette approche, nous présentons un outil automatique de visualisation et d'observation de la prolifération de cellule souche neuronale sous microscope à contraste de phase.The study of stem cells is one of the most important fields of research in the biomedical field. Computer vision and image processing have been greatly emphasized in this area for the development of automated solutions for culture and observation of cells. This work proposes a new methodology for observing and modelling the proliferation of neural stem cell under a phase contrast microscope. At each time lapse observation performed by the microscope during the proliferation, the system determines a three-dimensional model of the structure formed by the observed cells. This is achieved by a framework combining analysis, synthesis and selection process. First, an analysis of the images from the microscope segments the neurosphere and the constituent cells. With this analysis, combined with prior knowledge about the cells and their culture protocol, several 3-D possible models are generated through a synthesis process. These models are finally selected and evaluated according to their likelihood with the microscope image using a 3-D to 2-D registration method. Through this approach, we present an automatic visualisation tool and observation of the proliferation of neural stem cell under a phase contrast microscope.PARIS-JUSSIEU-Bib.électronique (751059901) / SudocSudocFranceF

    Uncertainty Minimization in Robotic 3D Mapping Systems Operating in Dynamic Large-Scale Environments

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    This dissertation research is motivated by the potential and promise of 3D sensing technologies in safety and security applications. With specific focus on unmanned robotic mapping to aid clean-up of hazardous environments, under-vehicle inspection, automatic runway/pavement inspection and modeling of urban environments, we develop modular, multi-sensor, multi-modality robotic 3D imaging prototypes using localization/navigation hardware, laser range scanners and video cameras. While deploying our multi-modality complementary approach to pose and structure recovery in dynamic real-world operating conditions, we observe several data fusion issues that state-of-the-art methodologies are not able to handle. Different bounds on the noise model of heterogeneous sensors, the dynamism of the operating conditions and the interaction of the sensing mechanisms with the environment introduce situations where sensors can intermittently degenerate to accuracy levels lower than their design specification. This observation necessitates the derivation of methods to integrate multi-sensor data considering sensor conflict, performance degradation and potential failure during operation. Our work in this dissertation contributes the derivation of a fault-diagnosis framework inspired by information complexity theory to the data fusion literature. We implement the framework as opportunistic sensing intelligence that is able to evolve a belief policy on the sensors within the multi-agent 3D mapping systems to survive and counter concerns of failure in challenging operating conditions. The implementation of the information-theoretic framework, in addition to eliminating failed/non-functional sensors and avoiding catastrophic fusion, is able to minimize uncertainty during autonomous operation by adaptively deciding to fuse or choose believable sensors. We demonstrate our framework through experiments in multi-sensor robot state localization in large scale dynamic environments and vision-based 3D inference. Our modular hardware and software design of robotic imaging prototypes along with the opportunistic sensing intelligence provides significant improvements towards autonomous accurate photo-realistic 3D mapping and remote visualization of scenes for the motivating applications
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