Thèses sur le sujet « Radiation-induced fibrosi »

La fibrose radio-induite (FRI) est une complication tardive de la radiothérapie souvent associée à une réponse inflammatoire chronique et à un infiltrat de macrophages. Aujourd’hui, les macrophages sont pressentis comme des médiateurs cellulaires important dans le processus de fibrose mais leur rôle n’a jamais été étudié dans le contexte de la FRI. Dans une précédente étude nous avions montré que l’irradiation (IR) induit une polarisation M1 des macrophages cardiaques après irradiation de souris ApoE-/- et est associée un score de fibrose élevé, ce qui suggérait que la polarisation des macrophages pourrait contribuer à la fibrogénèse radio-induite. Afin de valider cette hypothèse, nous avons cherché à caractériser le rôle des macrophages dans la FRI en utilisant un modèle classique de fibrose pulmonaire chez la souris C57Bl/6 induit après IR thoracique à 16Gy. Nous avons caractérisé les phénotypes et la fonction des macrophages alvéolaires (MA) et interstitiels (MI). Durant la phase précoce, les résultats montrent une déplétion des MA accompagnée de la sécrétion de CXCL1, MCP-1 et de MCSF. Cette déplétion est suivie d’une repopulation suite au recrutement et à la prolifération des monocytes/macrophages d’origine médullaire. La nouvelle population de MA présente une polarisation hybride accompagnée d’une augmentation simultanée de la sécrétion de cytokines Th1 et Th2. Durant la phase tardive les MI présentent une polarisation de type M2 accompagnée d’une diminution des cytokines Th1 et d’une augmentation de cytokines Th2 dans le lysat tissulaire. Nous avons ensuite cherché à caractériser la contribution des MA hybrides vs MI M2 dans le processus de fibrose. Nous avons montré que contrairement au MA hybrides, les MI M2 étaient capables d’induire l’activation des fibroblastes in vitro et l’expression de TGF-β1. De plus, la déplétion des MA hybrides avec une administration intranasale de clodronate exacerbe la FRI et induit l’augmentation de l’infiltrat de MI M2. Ensuite, nous nous somme interrogés à la contribution du processus de fibrose dans la polarisation des macrophages. Après 24h de coculture entre fibroblastes irradiés et macrophages pulmonaires non irradiés, une sécrétion de cytokines telles que M-CSF et TIMP-1 qui pourraient stimuler l’activation des fibroblastes est observée. De plus, l’inhibition de la FRI avec de la pravastatine montre que l’inhibition de la fibrose est accompagnée d’une augmentation des MI M1 et d’une diminution des MI M2 dans le poumon. En résumé, nos résultats montrent une contribution opposée des Macrophages Alvéolaires et des Macrophages Interstitiels dans le processus de fibrose radio-induite ainsi qu’une contribution du processus de fibrose dans le type d’activation des Macrophages interstitiels formant ainsi une boucle d’activation fibrogénique chronique
Radiation-induced fibrosis (RIF) is a delayed complication of radiotherapy often associated with chronic inflammatory process and macrophage infiltration. Nowadays, macrophages are suggested to be important cellular contributors to fibrogenic process, but their implication in the context of RIF has never been investigated. In a previous study we have shown that irradiation (IR) induced the polarization of cardiac macrophages into M1 in ApoE-/- mice and was associated with a high fibrosis score in ApoE-/- mice, suggesting that macrophage polarization could drive tissue sensitivity to ionizing radiation. This observation prompted us to investigate the role of macrophages in RIF using a classical experimental model of lung fibrosis developed in C57Bl/6 mice after 16Gy thorax-IR. We profiled both alveolar macrophages (AM) and interstitial macrophages (IM). During the acute phase we found AM depletion associated with CXCL1, MCP-1 and M-CSF secretion, followed by a repopulation phase mediated by recruitment and proliferation of monocytes/macrophages from the bone marrow. Interestingly, the newly recruited AM exhibited a yet never described hybrid polarization (M1/M2), associated with the up-regulation of both Th1 and Th2 cytokines. At delayed times points, IM were M2-polarized and associated with downregulation of Th1 cytokines and upregulation of Th2 cytokines in tissue lysates. These results suggest a differential contribution of hybrid AM vs M2 IM to fibrogenesis. Interestingly, in contrast to activated hybrid AM, activated M2 IM were able to induce fibroblast activation in vitro mediated by an enhanced TGF-β1 expression. Therefore, specific depletion of hybrid AM using intranasal administration of clodrosome increased RIF score and enhanced M2 IM infiltration. We next evaluated if the fibrogenic process can in turn affect macrophage polarization. Interestingly, after coculture of irradiated fibroblast with non-irradiated pulmonary macrophages, secretion of cytokines such as M-CSF and TIMP-1, which can stimulate macrophage activation, was observed. Furthermore, RIF inhibition using pravastatin treatment showed that fibrosis inhibition was associated with a decrease in M2 IM accompanied by an increase in M1 IM, but had no effect on polarization of AM. These present study shows a dual and opposite contribution of alevolar versus intertitial macrophages in RIF and the contribution of the fibrogenic process to IM polarization, resulting thereby in a chronical fibrogenic loop

La fibrose radio-induite est une complication sévère et tardive de la radiothérapie. Plusieurs études cliniques ont montré que la combinaison pentoxifylline-vitamine E est un traitement sûr et efficace contre la fibrose. Cependant, les mécanismes moléculaires de son efficacité restent inexplorés. Nous avons montré l’efficacité de la combinaison pentoxifylline-vitamine E dans l’entéropathie radique dans une faisabilité clinique. En parallèle, en utilisant un modèle unique, in vitro, de cellules musculaires lisses intestinales primaires isolées des personnes atteintes de l’entéropathie radique, nous avons montré une synergie entre la pentoxifylline et l’analogue hydrophile de vitamine E (trolox) qui permet à l’association d’inhiber l’expression de TGF-β1 au niveau de l’ARN messager et de la protéine. Cette action inhibitrice intervient au niveau transcriptionnel et conduit à une inhibition conséquente des cibles de la voie de signalisation TGF-β1/Smad (Col Iα1, FN1, PAI-1, CTGF), alors qu’elle semble sans effet sur la voie de signalisation Rho/ROCK. Pour la première fois, dans ces cellules issues de l’entéropathie radique, nous avons montré une surexpression de miR-210 ; microRNA induit par l’hypoxie. L’association pentoxifylline-trolox inverse la surexpression de miR-210 aussi bien dans les conditions normoxique que dans les conditions hypoxiques. L’implication de miR-210 dans l’entéropathie radique n’a pas été préalablement étudiée, néanmoins nous avons montré qu’un inhibiteur de miR-210 diminue l’expression de Col Iα1 dans ce modèle. L’effet anti-fibrosant exercé par l’association pentoxifylline-vitamine E est partiellement induit par l’inhibition de la cascade TGF-β1. L’inhibition de miR-210 est un deuxième mécanisme potentiel nécessitant d’autres investigations. Cette étude renforce les essais clinique antérieurs en montrant in vitro une synergie entre pentoxifylline et vitamine E et permettant de proposer cette association en première ligne thérapeutique dans la fibrose radio-induite. De plus, miR-210 est proposé comme une possible cible thérapeutique pour traiter la fibrose radio-induite
Radiation-induced fibrosis is a serious late complication of radiotherapy. Pentoxifylline-vitamin E has proven effective and safe in clinical trials as treatment of fibrosis, while the molecular mechanism of its activity is yet unexplored. We showed efficacy of Pentoxifylline-vitamin E combination in radiation-induced enteropathy in a small clinical study. In parallel, using a unique in vitro model of primary smooth muscle cells isolated from intestinal samples isolated from humans with radiation enteropathy we showed that pentoxifylline and the hydrophilic analogous of vitamin E (trolox) synergize to inhibit TGF-β1 protein and mRNA expression. This inhibitory action is mediated at the transcriptional level and leads to subsequent inhibition of TGF-β1/Smad targets (Col Iα1, FN1, PAI-1, CTGF), while it has no effect on the Rho/Rock pathway. We have also demonstrated, for the first time, an overexpression of the hypoxia-induced microRNA miR-210 in the fibrotic cells. Pentoxifylline-trolox combination could reverse this miR-210 overexpression in normoxic and hypoxic conditions. While miR-210 has not been previously shown to be involved in radiation-induced enteropathy, we showed that miR-210 inhibitor could reduce mRNA expression of Col Iα1. The anti-fibrotic effect of combined pentoxifylline-vitamin E is at least in part mediated by inhibition of the TGF-β1 cascade. MiR-210 inhibition is another mechanism which needs further investigations. This study strengthens previous clinical data showing pentoxifylline-vitamin E synergy and supports its use as a first-line treatment of radiation-induced fibrosis. Also, it suggests miR-210 as a new potential therapeutic target for the treatment of this complication

La radiothérapie thoracique peut induire le développement de pneumopathies aiguës et de fibroses. La dysfonction du système vasculaire participe au développement de lésions radiques. Dans l'intestin, un KO endothélial de PAI-1 protège les souris de la fibrose radique. Le premier objectif de ce projet est d'explorer le rôle de PAI-1 dans l'apparition de la fibrose radique pulmonaire. L'irradiation thoracique de souris à 17 Gy altère sévèrement le parenchyme pulmonaire et l'analyse histologique révèle que l'invalidation de PAI-1 aggrave les lésions à 2 et 13 semaines. Cette invalidation ne protège donc pas les animaux des dommages radiques pulmonaires. L'organisation en parallèle du poumon permet d'envisager une tolérance à de fortes doses par fraction sur des petits volumes. Des irradiations en conditions stéréotaxiques ont donc été réalisées chez la souris. Les analyses histologiques montrent une déstructuration alvéolaire et un fort infiltrat inflammatoire au niveau de la zone cible. Un œdème est observable dans l'ensemble du poumon ipsilatéral deux semaines après irradiation. Le poumon ipsilatéral est également affecté par des altérations de structure, tel un épaississement des septa alvéolaires. Ces bouleversements se traduisent également au niveau transcriptomique. A la vue de l'ensemble de ces altérations, un test à l'effort a été réalisé pour évaluer l'impact potentiel sur la fonction pulmonaire. Les résultats mettent en évidence une diminution des performances des animaux. Les analyses sont à approfondir mais elles démontrent l'importance de s'intéresser aux tissus sains situés hors du volume cible mais recevant des fractions variables de la dose délivrée
Radiation-induced endothelial dysfunction is known to participate to the development of normal tissue damage. PAI- is implicated in the phenotypic changes of irradiated endothelial cells and KOendo mice are protected from radiation damage to the gut. Whole thorax of PAI-1 KOendo and floxed mice were exposed to 17 Gy. Histological analyzes showed that PAI-1 KOendo induces a worsening of injuries at 2 and 13 weeks. Consequently, contrary to the gut no protection from radiation-induced lung damage is observed in PAI-1 KOendo mice. Our second aim was to study the effects of a single high dose stereotactic irradiation on pulmonary tissues. Histological analyzes and scanner imaging show important injuries on the targeted volume. An ipsilateral edema can also be observed 2 weeks after irradiation. Ipsilateral lung is moreover importantly damaged. A thickening of alveolar septa is notably observable. A transcriptomic analysis show important similarities between tissues from the ipsilateral lung and the focal lesion. As really highly damages have been observed in both scanner and histological analyzes, we decided to perform forced physical activity test on treadmill. A drastic decrease of maximal distance traveled has been observed from two weeks. These experiments highlighted a deficiency in respiratory function and all of these results show the importance of non-targeted irradiated pulmonary volume in the development of radiation-induced fibrosis. Effect of an endothelium-specific deletion of HIF-1α has been investigated in this model of stereotactic irradiation. Only few differences have been observed between KOendo and control mice. Experiments are still ongoing

Les fibres dopées erbium couvrent de nombreuses applications, particulièrement dans le domaine des télécommunications terrestres et sous marines, avec les amplificateurs optiques. Aujourd’hui, il existe un réel intérêt pour l’industrie spatiale d’utiliser ces fibres dans les satellites. Cependant, pour utiliser leur potentiel, une qualification en milieu radiatif doit être effectuée, c'est justement l'objet principal de ce travail de thèse. Grâce au partenariat industriel avec Draka-Prysmian, nous avons accès à une grande diversité de fibres en termes de compositions chimiques : ceci nous permet d’étudier leur sensibilité aux radiations, et de comprendre le rôle essentiel des dopants et des codopants dans cette sensibilité. Une étude de celle-ci en temps réel, associée à une caractérisation pré et post-irradiation des fibres optiques, rend possible l'identification fine des défauts induits sous irradiation, et la compréhension de leur mécanisme de formation, en fonction de la composition de ces fibres. Cette étude permet ainsi de proposer un modèle physique de leur dégradation, et aussi de leur guérison, complété par un modèle d'amplificateur. Il permet de prédire, en fonction de la composition des fibres, le comportement quantitatif des amplificateurs optiques associés, en termes de gain et et de bande passante, versus un dépôt de dose typique d'une mission spatiale ; il répond ainsi aux attentes des principaux acteurs du domaine. En outre, le bénéfice de ce travail ouvre des portes dans le domaine de la dosimétrie par fibre optique active, dans différents environnements radiatifs autres que le domaine spatial, tels que le milieu médical ou l'environnement nucléaire
Erbium-doped optical fibers open up many applications, especially in the field of terrestrial and underwater telecommunications, with optical amplifiers. Nowadays, there is a real interest for the space industry to use these fibers in satellites. However, in order to use their full potential, qualification in radiative environments is to be carried out, this is the main focus of this PhD work. Thanks to the partnership with Draka-Prysmian group, we have a full access to a large diversity of specialty fibers, in terms of chemical compositions : this allows us to study their sensitivity to radiations, and to determine the important role of dopants and co-dopants in this sensitivity. A real-time study of it, associated with a qualification of pristine and irradiated optical samples, enables the detection of radiation-induced defects, and the understanding of their creation process, as a function of the fiber structure. This study provides a physical model describing the degradation and the recovery of these fibers, enhanced with an amplifier modeling. It allows the prediction of the quantitative behavior of specialty fiber-based amplifiers, in terms of gain and bandwidth, versus the chemical composition of the fibers used, for a typical space mission dose ; thus this modeling meets the needs of the spatial market key actors. Furthermore, the benefit of this work opens up another avenues for some larger opportunities, in various radiative environments, such as the medical field or the areas of nuclear facilities

Dans ce travail de thèse, nous avons étudié l'effet des rayonnements ionisants (rayons X et γ) jusqu'à une dose maximale de 1 Grad sur différents types de fibres multimodes (dopées -P, -P-Ce, -Ge, -Ge-F, -Ge-Ce et -N). Les caractérisations ont été réalisées principalement avec trois techniques expérimentales : online Atténuation Induite par Radiation en temps réel (RIA), Résonance Paramagnétique Electronique (EPR), Micro-Luminescence (ML). Dans la première partie du travail de cette thèse, nous avons étudié la réponse aux radiations de différents types de fibres optiques. L'absorption liée aux défauts du phosphore induits par irradiation a été étudiée par des mesures RIA dans le domaine spectral UV-Visible. Les mesures EPR nous permis de détecter les défauts POHC, P1 et P2. En particulier, pour la détection de P1 et P2, nous avons utilisé le mode de détection de la seconde harmonique pour déterminer la cinétique de croissance des P1 et P2 en fonction de la dose. Nous avons également étudié les effets dus au changement des conditions de fibrage et ceux liés à la variation de la température d'irradiation (25-280 ° C). Nous avons aussi étudié l’effet du codopage du coeur de la fibre avec du Cérium. Dans ce cas, nous avons observé une production moins importante de centres POHC et P2 sous irradiation. De plus, les mesures EPR ont montré que la génération des défauts P1 n’est pas sensiblement influencée par le codopage avec le Cérium. En ce qui concerne les fibres optiques dopées Ge, on a étudié trois types de dopage : Ge seul, co-dopage Ge-F et Ge-Ce. Pour chaque type, nous avons examiné trois conditions de fibrage. La réponse à l’irradiation de ces fibres a été étudiée par les trois techniques utilisées. Plus particulièrement la ML, nous a permis d'obtenir une vision plus complète du rôle du codopant et des précurseurs dans la formation des défauts induits par l'irradiation. Nous avons également étudié la réponse au rayonnement de la fibre dopée N avec trois différentes conditions de fibrage. Les réponses à l’irradiation dans les régions spectrales UV-visible ont été obtenues par des mesures RIA. Par EPR, nous avons pu détecter deux défauts liés à l'azote pour les doses élevées de radiation. Enfin, les mesures ML sur les fibres irradiées ont montré trois bandes d'émission dans le visible qui ont été attribuées clairement à des centres émetteurs liés à l'azote. Dans la deuxième partie de la thèse, nous avons étudié les effets liés au chargement en oxygène des fibres étudiées. Par des mesures en microspectroscopie Raman, nous démontrons qu'un traitement à haute température et haute pression peut favoriser l’introduction d’une grande quantité de O2 dans les fibres optiques à cœur de silice pure (PSC) ou dopées F, Ge ou P. Les réponses à l’irradiation de certaines des fibres optiques chargées en O2 ont été étudiés (et en particulier PSC et celle dopée F. Sur la base des données de la littérature, nous avons effectué les décompositions des spectres RIA en fonction de la dose. De plus, l'étude EPR des fibres optiques dopées P et chargées en O2 a montré une forte réduction des défauts P1 et P2 comparées aux fibres non traitées. Dans cette partie de la thèse, j’ai également présenté les résultats concernant la radioluminescence infrarouge (1272 nm) des molécules O2 dans la fibre optique. La faisabilité d'un capteur de radiation pour des environnements sous fortes doses et forts débits de dose a été discutée
In this Thesis work we have investigated the effect of ionizing irradiation (X and γ rays) up to 1 Grad on different types of multimode optical fibers (P-doped, P-Ce-doped , Ge-doped, Ge-F-doped, Ge-Ce-doped, and N-doped). The experiments were carried out by three main experimental techniques: online Radiation Induced Attenuation (RIA), Electron Paramagnetic Resonance (EPR) and Confocal Micro-Luminescence (CML). In the first part of the Thesis work we report on the radiation response of several types of optical fibers. The absorption due to radiation induced P-related defects was studied by RIA in the UV-Visible domain. Moreover, by EPR measurements we were able to detect POHC, P1 and P2 defects. In particular, for the detection of P1 and P2 defects we have validated the use of EPR second-harmonic detection mode which allowed us to obtain the growth kinetics of P1 and P2 with the dose. The effects due to the variation of the drawing conditions of the fibers were investigated as well as the ones due to the change of the temperature of irradiation (from 25 to 280 °C). Finally, concerning the P-doped OFs, we report on the effects due to the Cerium codoping of the core of the optical fiber. We observed a reduced generation of POHC and P2 centers under irradiation. However, EPR investigation has shown that the generation of P1 defects is essentially unaffected by the Ce-codoping. Regarding Ge-doped optical fibers we report on three basic typologies: Ge-doped, Ge-F-doped and Ge-Ce-doped. For each fiber typology we investigated three drawing conditions. The radiation responses of these fibers were characterized by RIA and EPR measurements. Furthermore, performing CML measurements we were able to obtain further insight on the role of the co-dopants and of the defect precursors in determining the radiation induced defects. We have also investigated the radiation response of N-doped OFs (three drawing conditions). The radiation responses in the UV-Visible domains were obtained by RIA, and by EPR measurements we were able to detect the signals of two N-related defects at high radiation doses. Finally, CML measurements on irradiated samples have shown three emission bands in the visible domain which are tentatively assigned to N-related centers. In the second part of the Thesis we report on the effects of an O2 loading treatment produces on some of the investigated samples. By micro-Raman measurements we demonstrate that a high pressure high temperature treatment can incorporate high quantity of O2 into Pure-Silica-Core (PSC), F, Ge and P doped optical fibers. The radiation responses of some of the O2-loaded optical fibers were investigated with particular regard to the fluorine doped and pure-silica-core optical fibers. On the basis of literature data we performed band decompositions of the RIA spectra as a function of the dose. Moreover, the EPR study of the O2 loaded P-doped optical fiber have shown a strong reduction of the signals associated to the P1 and P2 defects as compared to the untreated fibers. In this part of the thesis we also report on the characterization of the near infrared radioluminescence (1272 nm) of O2 molecules embedded in the optical fiber matrix and the feasibility of a radiation sensor based on this phenomenon for environments characterized by high radiation doses and high dose-rates

EDF souhaite utiliser la technologie de mesure de température répartie par capteur à fibre optique utilisant l’effet Raman pour réaliser des cartographies de température de certains composants de centrales nucléaires. Les conditions environnementales auxquelles le capteur à fibre optique est soumis sont particulièrement agressives (température de 350 °C et rayonnements gamma ionisants). Les rayonnements ionisants sont responsables de la création de défauts structurels au cœur de la fibre, qui atténuent sa transmission lumineuse, et dont les effets engendrent une erreur de mesure de température pouvant aller jusqu'à l’interruption totale de la mesure. La haute température, quant à elle, dégrade le revêtement protecteur de la fibre optique, ce qui la fragilise mécaniquement. Des irradiations gamma in situ sur des fibres optiques multimodes commerciales à revêtement or protégées par une gaine en acier inoxydable ont été réalisées, à l’aide de deux sources de rayonnements différentes, pour observer l'atténuation radio-induite du capteur à fibre optique en fonction du débit de dose et de la dose cumulée. Les effets du rayonnement à température ambiante, puis à haute température, ont été observés. Ce travail expérimental démontre que la haute température peut être maîtrisée grâce à une fibre à revêtement or, et que la haute température est bénéfique contre l’atténuation de la fibre engendrée par l’irradiation. La mise en œuvre de capteur de température à fibre optique en environnement sévère devient possible, ainsi que l’estimation des incertitudes sur la mesure associée
EDF is working on Raman distributed temperature sensing using optical fiber sensors in order to map temperature of nuclear power plants big components. The sensor has to sustain harsh environmental conditions (temperatures up to 350 °C and gamma ionizing radiations). Ionizing radiations can create structural defects inside the fiber’s core, which attenuate the light transmission. This phenomenon can lead to temperature measurements errors until no measurement is possible. As for high temperature, it can affect the fiber coating, which mitigate the fiber mechanical resistance.Gamma rays in situ irradiations have been carried out over commercial off-the-shelf multimode gold coated fibers protected with a stainless steel metal tubing, with two different radiation sources, in order to observe radiation-induced attenuation over dose rate or cumulated dose. Effects of gamma rays over gold coated optical fiber sensors have been observed at both room anhigh temperature.This experimental work enlightens that high temperature can be controlled with gold coated fibers, and that the radiation-induced attenuation downsides can efficiently be balanced with high temperature. Implementation of a Raman distributed temperature optical fiber sensor in such harsh environments becomes possible, as well as the associated estimation of measurement uncertainty

Cette thèse est consacrée à l'étude de la réponse aux radiations d'amplificateurs à fibre optiques dopées Er 3+ et Yb3+. Ces dispositifs fonctionnant à 1,5 µm ont été conçus pour des applications spatiales et l'évaluation de leurs performances revêt d’une importance capitale dans un tel environnement hostile. Deux traitements, le chargement en H2 et le co-dopage au Ce du cœur de la fibre, ont été étudiés comme solutions de durcissement aux radiations. Une étude spectroscopique a permis d’approfondir la connaissance des mécanismes physiques de base responsables de la dégradation de ces composants et par conséquent de proposer des solutions de durcissement. La thèse est organisée en trois parties. La Partie I présente une description générale des fibres dopées aux ions de Terres Rares (TR), avec l'introduction des concepts de base de la physique de tels éléments et leur interaction avec la matrice hôte (verre phosphosilicate). L'état de l'art concernant les effets des rayonnements sur les fibres dopées aux TR est également présenté. La Partie II décrit les échantillons et les techniques expérimentales utilisées. La Partie III décrit les principaux résultats dont les tests, en configuration active, démontrent que le co-dopage au Ce ainsi que le chargement en H2 ont un rôle-clé dans la limitation des pertes induites par rayonnement. L'analyse spectroscopique de la matrice vitreuse (Raman) et des ions TR (par mesures de luminescence stationnaire et résolue en temps) mettent en exergue un fort effet de durcissement, conduisant à une préservation de l'efficacité du système physique en opération
This thesis is devoted to the study of the radiation response of optical amplifiers based on Er/Yb doped fibers. These devices operating at 1.5 µm are conceived for space applications and contextually the evaluation of their performance in such harsh environment becomes of crucial importance. Two treatments, the H2-loading and the Ce-doping of the fiber core, are investigated as radiation hardening solutions. A spectroscopic study has been associated, in order to improve the knowledge of the physical mechanisms responsible for the signal degradation and the action of the hardening solutions. The thesis is organized in three parts. Part I deals with a general description of the Rare-Earth (RE)-doped fibers, with the introduction of some basic concepts of the RE-ion physics and their interaction with the host matrix material (phosphosilicate glass). The state-of-art of the radiation effects on the optical fibers, particularly the RE- doped fibers, is also overviewed. Part II describes the samples (fiber fabrication, geometry and chemical compositions), and the used experimental techniques, including a short discussion on the related theoretical background. Part III describes the main results; firstly, the active tests, performed on the RE-doped fiber as part of an optical amplifier, demonstrate that the Ce-codoping and H2-load have a key-role in the limitation of the radiation induced losses. Then, the spectroscopic analysis of the phosphosilicate glass (Raman study) and of the RE-ions (stationary and time-resolved luminescence) show a stabilization effect due to the two treatments, leading to a preservation of the high efficiency of the physical system under study

Radiation-induced fibrosis (RIF) is one of the common adverse effects of radiotherapy (overall incidence between 10-20%) and it results in a multitude of symptoms that can significantly impact the quality of life of cancer patients. RIF is a complex multi-factorial process that ultimately results in an excess accumulation of collagen and other extracellular matrix components in many organs, especially in the lung, skin, small bowel, breast, liver, and kidney, that in turn impairs their own functions. RIF is initiated by direct radiation-induced cell damage and by indirect mechanisms which, in turn, involve the generation of reactive oxygen and nitrogen species (ROS and RNS, respectively) through water radiolysis and activation of nitric oxide synthase. Central to RIF is the role of tissue-resident leukocytes, such as macrophages, which, in response to ROS accumulation and tissue damage, activate the expression of pro-inflammatory cytokines, such as transforming growth factor beta (TGF-β). TGF-β may, in turn, increase ROS production through suppression of antioxidant enzymes, thus leading to a positive feedback that sustains oxidative stress, inflammation and tissue damage. f-β, finally, promotes fibroblasts recruitment and local deposition of extracellular matrix components. Our understanding of the processes involved in the pathology of RIF have improved. However, to date no successful therapeutic approaches are available in the clinical setting. Importantly, it has been reported that post- radiation antioxidant therapy significantly reduces RIF in animal models. This shows that it is indeed possible to halt the positive feedback between ROS accumulation and inflammation at the basis of the disease. Astaxanthin (3,3′-dihydroxy-β-β′-carotene-4,4′-dione; ASX) is a xantophyll ketocarotenoid present in several marine and freshwater organisms, including microorganisms, crustaceans and fishes. ASX is a potent antioxidant, with rate constants for radical scavenging approximately 10 times higher than other carotenoids, such as lutein and β-carotene, and 100 times more than that of α- tocopherol. ASX might, therefore, be used to reduce or prevent RIF. ASX, however, is not soluble in water-based biological fluids and is sensitive to temperature, light and oxygen, all factors that limit its stability and bioavailability and that can be controlled by ASX encapsulation into appropriate chemical matrices. Encapsulation of ASX into micrometer-sized particles might also represent an excellent strategy to deliver ASX specifically to macrophages and other innate immune cells that are characterized by their unique ability to engulf, -5- uptake and degrade particles of this size through phagocytosis. In contrast, tumor cells do not phagocytose and hence will not be provided direct protection against antioxidants conferred by ASX. This project aims at demonstrating the feasibility of this possible therapeutic approach against RIF, albeit in a cell-based in vitro setting. The results clearly indicate that ASX microparticles interfere with the positive feedback between ROS and TGF-β in phagocytic cells, and that pentoxifylline, a drug currently investigated for its ability to reduce RIF, amplifies the effects of astaxanthin microparticles on macrophages. Treatment strategies involving ASX microparticles, eventually in combination with other drugs such as pentoxifylline, should be further studied to examine their potential to reduce inflammation and inhibit radiation-induced fibrosis.

碩士
國立臺灣大學
臨床醫學研究所
104
Radiation-induced pulmonary fibrosis (RIPF) is a late side effect of thoracic radiotherapy with few effective treatments available. Fibrosis progression not only impedes further radiation treatment but also brings life-threatening condition. The pathophysiology of RIPF is not well understood, but elevated serum levels of IL-6 is a reliable marker for disease severity. Inhibition of IL-6 results in attenuation of pulmonary fibrosis in mice. However, it is unclear whether this is due to blockade of classical signaling, mediated by membrane-bound IL-6Rα(mIL-6Rα), or trans-signaling, mediated by soluble IL-6Rα (sIL-6Rα). Here, we assessed the role of sIL-6Rα in RIPF. We demonstrated activation of IL-6 trans-signaling in mice during the onset and progression of lung fibrosis. Using a mouse model of RIPF, we demonstrated that sgp130Tg mice had a remarkable reduction in RIPF and a lower death rate compared with wild type C57BL/6 mice. This observation was associated with an attenuation of pulmonary epithelial-to-mesenchymal transition (EMT) in sgp130Tg mice. In vitro, IL-6 trans-signaling stimulated phosphorylation of STAT3 and suppressed of E-cadherin expression in lung epithelial cells, effects relevant in the progression of pulmonary fibrosis. Taken together, these results provide the first evidence that IL-6 trans-signaling mediated signaling cascade plays an essential role in the pathogenesis of RIPF, and suggest that selective inhibition of IL-6 trans-signaling may be a novel therapeutic strategy for the management of RIPF.

Radiation-induced lung injury (RILI) is a common condition in the setting of lung and breast cancer. Often, patients who suffer from RILI experience pneumonitis and pulmonary fibrosis months after treatment. These pathologies have commonly been modeled using mice and observing their deterioration until mortality and quantifying pathology on histological sections.

With this study, we used a longitudinal microCT and a 7T MRI to characterize male C57Bl/6 mice irradiated with a single dose of 20 Gy to the whole thoracic area delivered by an X-Rad cabinent irradiator. CT was performed with a respiratory gating sequence at 2 week timepoints to construct an RIPF model. The fraction of RIPF to total lung volume was calculated at each time point from images, and the data was anaylzed using one-way ANOVA Welch and Dunnett’s T3 multiple comparisons tests. Tidal lung volumes were also calculated and anlyazed in a simlar manner. Mice were then imaged using MRI and CT at 0, 5, and 8 week timepoints to compare results. These results were analyzed for comparison (ANOVA and Dunnett’s T3) and correlation (Pearson’s r) with each other. Histology was later performed using H&E and Trichrome stains to provide ex-vivo verification of pathology. At the 10-12 week time point ( ) significant RIPF formed. Weeks proceeding showed increased significance until the 22+ week timepoint, which showed less statistical significance ( ) due to increased variance at this timepoint. Dunnett’s T3 test showed no significant differences between tidal lung volumes over time. Tests also showed no significant differences between CT and MRI results with a correlation coefficient of . Early in the study, problems arose when pre-marture mortality was occurring to a significant portion of our subjects. Analysis later showed issues during irradiation that resulted in significant dose being absorbed by the stomach. Adjusting our shiedling lead to increased early survival of our subjects enabling us to contine our study. Significant RIPF development was not significant until 10-12 weeks post-irradiation, then RIPF became more severe at proceeding timepoints. Tidal lung volume showed no significant deviation over the development of RIPF. This result is most likely affected by the variation of results at later timepoints, since several mice with severe RIPF were significantly hindered in their ability to breathe during the study. MRI results showed close correlation with CT results and prodcued similar values at early timepoints. However, noticeable differences were seen at later timepoints when significant RIPF developed ( ).

碩士
國立臺灣海洋大學
食品科學系
106
Radiation-induced fibrosis (RIF) is one of the most common late complications of radiation therapy. This study was aimed to evaluate the possible protective effects of Low molecular weight fucoidan (LMF) against γ-radiation induced damage in vitro. It was observed that the exposure to γ-radiation dose-dependently caused a significant decrease in the cell viability. The pre-treatment and pos-treatment of LMF attenuated 1 Gy cytotoxicity. In addition, LMF also reduce γ-irradiation induced fibrosis responses, as indicated by the decreased production of TGF-β1, smad3, collagen-I, fibronectin and α-SMA signaling pathway has been reported to be a key mediator involved in γ-radiation-induced cellular fibrosis. TGF-β1 is a master regulator of γ-irradiation induced fibrosis. Thus, we use TGF-β1 proteins induced cellular fibrosis to determine the effects of LMF on fibrosis index and cellular morphology. LMF pre-treatment significantly reduced p-smad3 active and form heteromeric complexes with smad 4, collagen accumulation in cell fibrosis. The long/short axis ratio was evaluated to assess the morphological differences. It was observed that the exposure to TGF-β1 caused a significant decrease in the average ratio. The pre-treatment of LMF dose-dependently caused increase average ratio. We found that LMF effectively prevents RIF, which might be a new therapeutic methods that can be used in the prevention-treatment of RIF.

碩士
高雄醫學大學
牙醫學系碩士班
104
Oral squamous cell carcinoma (OSCC) is one of the most diagnosed malignancy of head and neck regions with growing morbidity, and mortality, while RIF (radiation induced fibrosis) is one of the most severe long-term effects of radiotherapy for oral cancer. Like other fibrotic diseases, which may be exaggerated for years even whole life. Present study was to investigate TGFβR1 (transforming growth factorβ receptor 1) and its’ potential downstream mediators in field of OSCC and fractionated RIF in DMBA (7, 12-dimetheyl benz[a]anthrance) induced hamster buccal pouch squamous cell carcinoma model. Thirty-six male golden Syrian hamsters were divided into five groups: DMBA + Radiation (DR), DMBA (D), Radiation (R), mineral oil (MO) and no treatment (N). Hamsters were induced buccal pouch cancer by topical application of DMBA for 12 weeks, and all the hamsters in the DR and RT groups received whole head irradiation by the linear accelerator with a total dose of 42Gy divided into 6 fractions, twice weekly. Immunohistochemistry and western blotting studies showed that TGFβR1 (transforming growth factorβ receptor 1), Smad3 (drosophila mothers against decapentaplegic protein 3), Erk1/2 (extracellular signal-regulated kinase 1/2), CCN2 (connective tissue growth factor) were highly expressed in cancer cells. The gross pouch tissues in the D & DR groups showed thickened and significant volume size shrinkage, histopathology/immunohistochemistry studies showed that D and DR groups were highly fibrotic, TGFβR1, Smad3, p-Smad3, Erk1/2, p-Erk1/2, CCN2 and α-SMA (α-smooth muscle actin) expressions were all statistically significantly increased when compared with N and R groups. Only Smad3, p-Smad3, and Erk1/2 were higher in DR group than in D group with marginally statistical significant (p=0.051). In conclusion, present study indicated TGFβR1, Smad3, Erk1/2, and CCN2 may be crucial in hamster buccal pouch OSCC and RIF. Though there was no direct evidence of association within upregulation of above proteins in our experiment, further study may be conducted toward these targets. DMBA-induced hamster buccal pouch cancer model could be employed to the study of OSCC and its radiation effects.