Submitted by Nadir Basilio (nadirsb@uninove.br) on 2016-05-19T18:09:00Z No. of bitstreams: 1 Rubia Garcia Lopes.pdf: 6178642 bytes, checksum: 787a2ca51a971a3c09bda8ef659636e1 (MD5) / Made available in DSpace on 2016-05-19T18:09:00Z (GMT). No. of bitstreams: 1 Rubia Garcia Lopes.pdf: 6178642 bytes, checksum: 787a2ca51a971a3c09bda8ef659636e1 (MD5) Previous issue date: 2014-12-11 / Halitosis is a term used to define the unpleasant breath that may have a systemic or oral origin. Volatile sulfur compounds produced by the Gramnegative bacteria mainly cause the bad breath. Using light - along with by chemical agents or not - is common to induce therapeutic and antimicrobial effects in the photodynamic therapy, and the antimicrobial effect happens only in the areas covered by the dye and irradiated by light. The aim of this study was to evaluate the antimicrobial effect of the photodynamic therapy in adolescent halitosis, analyzing the volatile sulfur compounds concentration, measured by gas chromatography (OralChromaTM). 45 adolescents were assessed and randomly divided (through controlled clinical study) into 3 groups that received different treatments: group 1 treatment with photodynamic therapy applied on the back (dorsum) region and on the middle third of the tongue, group 2 tongue scraper and group 3 treatment with tongue scraper and photodynamic therapy. The halitosis diagnosis was performed before and after the OralChroma treatment. The Kruskal-Wallis test was applied and compared with the Student-Newman-Keuls test. The α = 0.05 significance level was considered for all analysis. After the treatment, there was a statistically significant decline on all groups (p <0.001); however, the photodynamic therapy and tongue scraper treatment proved to be more efficient to fully reduce the hydrogen sulfides (median = 0). This study provides a new option for treating adolescent halitosis with immediate effects without mechanical aggression to the lingual papillae, which is common in the conventional treatment with tongue scrapers. / A halitose é um termo utilizado para definir o odor desagradável e fétido que emana da boca, podendo apresentar origem sistêmica (10%) ou oral (90%). O mau odor é provocado principalmente por compostos sulforados voláteis, produzido pela ação de bactérias Gram-negativas. A luz acompanhada ou não de agentes químicos tem sido usada para induzir efeitos terapêuticos e antimicrobianos na terapia fotodinâmica (TFD), sendo que o efeito antimicrobiano fica confinado apenas às áreas cobertas pelo corante e irradiadas pela luz. O objetivo deste estudo foi avaliar o efeito antimicrobiano da TFD em adolescentes com halitose, pela análise da concentração de compostos sulforados voláteis, mensurado por cromatografia gasosa (OralChroma TM). Por meio de estudo clínico controlado, 45 adolescentes foram avaliados e divididos aleatoriamente em 3 grupos que receberam tratamentos distintos: grupo 1 tratamento com TFD aplicada na região de dorso e terço médio da língua, grupo 2 raspador lingual e grupo 3 tratamento combinado de raspador lingual e TFD. O diagnóstico de halitose foi realizado antes e depois do tratamento pela cromatorgrafia gasosa. Foi aplicado o teste de Kruskal-Wallis para comparação seguido do teste Student-Newman-Keuls. Para todas as análises foi considerado um nível de significância de α=0,05. Após o tratamento houve redução estatisticamente significante para todos os grupos (p < 0,001), contudo a associação da terapia fotodinâmica ao raspador lingual mostrou ser mais eficiente na redução total de sulfidretos (mediana=0). Conclui-se portanto, que esse estudo traz uma nova opção de tratamento para halitose, com efeito imediato e sem agressão mecânica as papilas linguais comum ao tratamento convencional com raspadores.

Objetivo: Evaluar el efecto inhibitorio de la terapia fotodinámica (TPD) con Diodo Emisor de Luz (LED) azul sobre Streptococcus mutans y Streptococcus sanguinis en presencia y ausencia de riboflavina (E - 101). Materiales y métodos: Se realizaron cuatro tratamientos en presencia y ausencia de la exposición de luz LED azul y riboflavina al 0.5% sobre Streptococcus mutans y Streptococcus sanguinis. Las bacterias fueron cultivadas en medio BHI y la unidad de medida utilizada fue las unidades formadoras de colonias (UFC/ml). Resultados: La fotoactivación con luz LED azul a 40 segundos no tuvo efecto inhibitorio sobre S. mutans y S. sanguinis. Sin embargo, al realizar la terapia fotodinámica en presencia de riboflavina, se observó que el crecimiento bacteriano fue menor (p<0.05). Asimismo, se identificó que la viabilidad bacteriana de S. sanguinis es menor que la de S. mutans, con un 40% y 66% respectivamente. Conclusiones: Se concluye que la riboflavina tiene un efecto inhibitorio significativo sobre la viabilidad bacteriana de S. mutans y S. sanguinis. / Objective: To evaluate the inhibitory effect of photodynamic therapy (TPD) with blue Light Emitting Diode (LED) on Streptococcus mutans and Streptococcus sanguinis in presence and absence of riboflavin (E-101). Materials and methods: Four treatments were performed in presence and absence of blue LED and riboflavin (0.5%) exposure on Streptococcus mutans and Streptococcus sanguinis. The bacteria were grown in BHI medium and the unit of measurement used was the colony forming units (CFU / ml). Results: Photoactivation with blue LED light at 40 seconds had no inhibitory effect on S. mutans and S. sanguinis. However, when performing photodynamic therapy in presence of riboflavin, it was observed that bacterial growth was lower (p <0.05). Likewise, it was identified that bacterial viability of S. sanguinis is lower than S. mutans, with 40% and 66% respectively. Conclusions: It is concluded that riboflavin has a significant inhibitory effect on the bacterial viability of S. mutans and S. sanguinis. / Tesis

Le rétinoblastome est la tumeur intraoculaire primitive la plus fréquente de l’enfant. Les traitements actuels du rétinoblastome sont associés à de nombreux effets secondaires. De nouvelles approches thérapeutiques (telles que la photothérapie dynamique [PDT] ou les injections intra-vitréennes [IVT] de chimiothérapies) doivent donc être évaluées sur des modèles animaux, en vue d’une éventuelle application clinique.Dans cette thèse nous avons tout d’abord caractérisé un modèle murin obtenu par xénogreffe orthotopique de cellules issues de rétinoblastomes humains. Nous avons montré que la croissance tumorale intraoculaire est possible dans des lignées de souris immunodéficientes (Swiss-nude et SCID [severe combined immunodeficiency]) et dans une lignée immunocompétente (B6Albino). En raison du taux de prise tumorale insuffisant (entre 28.4% et 68.8% selon les lignées de souris utilisées) et des complications oculaires liées à l’injection orthotopique de cellules tumorales (cataracte, décollement de rétine chronique), les tests thérapeutiques (PDT et IVT de chimiothérapies) ont ensuite été réalisés sur un modèle murin transgénique de rétinoblastome (LHBetaTag).En vue du traitement par PDT, une étude de biodistribution par IRM (imagerie par résonance magnétique) du photosensibilisateur (PS, DEG-mannose) couplé au manganèse et une étude par dosage du PS ont été réalisées. Elles ont toutes les deux montré que l’illumination de la tumeur doit être réalisée 24 à 48h après l’administration intra-péritonéale du PS (ce qui correspond au « drug-to-light interval » du traitement par PDT). En utilisant ces paramètres, le traitement par PDT a été efficace sur les tumeurs rétiniennes des souris LHBetaTag. Au niveau de la zone traitée par PDT, il y a ainsi eu 91.7% de cicatrices choriorétiniennes en OCT (optical coherence tomography) pour un « drug-to-light interval » de 24h et 100% de cicatrices choriorétiniennes pour un « drug-to-light interval » de 48h. La rétine non tumorale située en dehors de la zone traitée par PDT avait un aspect normal en histologie, ce qui est en faveur d’une absence de toxicité rétinienne de la PDT sur les tissus sains. Le traitement par laser seul n’a pas eu d’effet anti-tumoral.Des traitements par IVT de chimiothérapies ont aussi été évalués sur les tumeurs rétiniennes des souris LHBetaTag. Les molécules utilisées ont été le melphalan, le carboplatine et le topotecan, administrées en mono ou en bithérapie. Nous avons montré que 4 IVT hebdomadaires de carboplatine à la dose de 1.5 µg ont la meilleure efficacité anti-tumorale (83.3% d’yeux sans masse tumorale en histologie) pour une toxicité rétinienne faible (21.4% d’yeux où il y a eu une diminution de l’épaisseur de la rétine non tumorale en OCT au cours du suivi in vivo). Le carboplatine semble donc être une alternative intéressante au melphalan, qui est actuellement la molécule la plus utilisée en clinique pour les IVT dans le rétinoblastome mais qui est associé à une toxicité rétinienne importante.En conclusion, ces études précliniques réalisées sur un modèle murin de rétinoblastome (LHBetaTag) montrent que la PDT est envisageable pour le traitement des tumeurs rétiniennes dans le rétinoblastome humain. Les IVT de carboplatine sont une perspective pour le traitement des flocons intra-vitréens dans cette maladie. Des évaluations fonctionnelles (électrorétinogramme, étude du réflexe optocinétique) devront cependant être réalisées chez la souris avant un éventuel passage en clinique afin de mieux caractériser une éventuelle toxicité rétinienne de ces traitements. / Retinoblastoma is the most common primary intraocular malignancy in children. Current retinoblastoma treatments have many adverse effects. New therapeutic approaches (like photodynamic therapy [PDT] or intravitreal injections [IVT] of chemotherapy) must therefore be evaluated on animal models, before a clinical application.In this thesis we first characterized an orthotopic xenograft murine model obtained with human retinoblastoma cells. We showed that intraocular tumor growth can be achieved in immunodeficient mouse strains (Swiss-nude and SCID [severe combined immunodeficiency]) and in an immunocompetent strain (B6Albino). Due to insufficient tumor engraftment rates (between 28.4 and 68.8% depending on the mouse strains) and to ocular complications after the injection of tumor cells (cataract, chronic retinal detachment) the treatments (PDT and IVT of chemotherapy) were performed on a transgenic retinoblastoma mouse model (LHBetaTag).In order to perform PDT, an MRI study (magnetic resonance imaging) of the photosensitizer (PS, DEG-mannose) coupled with manganese and a biodistribution study based on the dosage of the PS were performed. Both studies showed that the illumination of the tumor should be performed between 24 and 48h after the intraperitoneal injection of the PS (which corresponds to the “drug-to-light interval” of PDT). Using these parameters, PDT was effective on the retinal tumors of LHBetaTag mice. In the area treated with PDT we found 91.7% chorioretinal scars on OCT (optical coherence tomography) with a “drug-to-light interval” of 24h, and 100% chorioretinal scars with a “drug-to-light interval” of 48h. The retina outside the treated area had a normal aspect on histology, showing that PDT is not toxic on healthy tissues. Laser treatment alone had no anti-tumor effect.IVT of chemotherapy were also performed in LHBetaTag mice. We used melphalan, carboplatin and topotecan, alone or in association. We showed that 4 weekly IVT of carboplatin at the dose of 1.5 µg had the best anti-tumor effect (83.3% of eyes had no tumor mass on histology) and little retinal toxicity (21.4% of eyes had diminished retinal thickness on OCT). Carboplatin seems an interesting alternative to melphalan which is currently the most commonly used chemotherapy for IVT (but has a retinal toxicity).In conclusion, these preclinical studies on a retinoblastoma mouse model (LHBetaTag) show that PDT could be used to treat retinal tumors in human retinoblastoma. IVT of carboplatin could be used to treat vitreous seeds in this disease. Functional tests (electroretinogram, optokinetic reflex) should be performed in mice in order to evaluate more precisely the retinal toxicity of these treatments.

Horimatsu T, Muto M, Yoda Y, Yano T, Ezoe Y, et al. (2012) Tissue Damage in the Canine Normal Esophagus by Photoactivation with Talaporfin Sodium (Laserphyrin): A Preclinical Study. PLoS ONE 7(6): e38308. doi:10.1371/journal.pone.0038308 / 京都大学 / 0048 / 新制・論文博士 / 博士(医学) / 乙第12870号 / 論医博第2086号 / 新制||医||1006(附属図書館) / 31588 / (主査)教授 羽賀 博典, 教授 坂井 義治, 教授 平岡 眞寛 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・論文博士 / 博士(医学) / 乙第13132号 / 論医博第2136号 / 新制||医||1024(附属図書館) / (主査)教授 溝脇 尚志, 教授 坂井 義治, 教授 山田 泰広 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM

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Two-photon absorbing (2PA) materials has been widely studied for their highly localized excitation and nonlinear excitation efficiency. Application of 2PA materials includes fluorescence imaging, microfabrication, 3D data storage, photodynamic therapy, etc. Many materials have good 2PA photophysical properties, among which, the fluorenyl structure and its derivatives have attracted attention with their high 2PA cross-section and high fluorescence quantum yield. Herein, several compounds with 2PA properties are discussed. All of these compounds contain one or two fluorenyl core units as part of the conjugated system. The aim of this dissertation is to discuss the application of these compounds according to their photophysical properties. In chapters 2 to 4, compounds were investigated for cell imaging and tissue imaging. In chapter 5, compounds were evaluated for photodynamic therapy effects on cancer cells. Chapters 2 and 3 detail compounds with quinolizinium and pyran as core structures, respectively. Fluorene was introduced into structures as substituents. Quinolizinium structures exhibited a large increase in fluorescence when binding with Bovine Serum Albumin (BSA). Further experiments in cell imaging demonstrated a fluorescence turn-on effect in cell membranes, indicating the possibility for these novel compounds to be promising membrane probes. Pyran structures were conjugated with arginylglycylaspartic acid peptide (RGD) to recognize integrin and introduced in cells and an animal model with tumors. Both probes showed specific targeting of tumor vasculature. Imaging reached penetration as deep as 350 µm in solid tumors and exhibited good resolution. These results suggest the RGD-conjugated pyran structure should be a good candidate probe for live tissue imaging. Chapter 4 applied a fluorene core structure conjugated with RGD as well. Application of this fluorenyl probe compound is in wound healing animal models. Fluorescence was collected from vasculature and fibroblasts up to ≈ 1600 µm within wound tissue in lesions made on the skin of mice. The resolution of images is also high enough to recognize cell types by immunohistochemical staining. This technology can be applied for reliable quantification and illustration of key biological processes taking place during tissue regeneration in the skin. Chapter 5 describes three fluorenyl core structures with photoacid generation properties. One of the structures showed excellent photo-induced toxicity. Cancer cells underwent necrotic cell death due to pH decrease in lysosomes and endosomes, suggesting a new mechanism for photodynamic therapy.

Photodynamic therapy (PDT) has been investigated over the past three decades and is currently an approved therapeutic modality for skin cancer, the treatment of superficial bladder, early lung and advanced esophageal cancers, and age-related macular degeneration in a number of countries. In PDT, the absorption of light by a chromophore generates cytotoxic species such as reactive singlet oxygen, leading to irreversible destruction of the treated tissue. The measurement of the singlet oxygen quantum yield is an important determinant used to evaluate the efficiency of new photodynamic therapy agents developed in the laboratory, to screen potential photosensitizers in aqueous media.The singlet oxygen quantum yield is a quantitative measurement of the efficiency in which photosensitizers are able to use energy, in the form of light, to convert oxygen in the ground state to the reactive species singlet oxygen useful in photodynamic therapy. Singlet oxygen quantum yields of photosensitizers differ when measured in different solvents. The majority of the existing quantum yield values found in literature for various photosensitizers are documented with the sensitizers in organic solvents though values in aqueous media are more valuable for actual applications. Determination of accurate and precise quantum yield values in aqueous solution is a much more difficult problem than in organic media. Problems in aqueous solution arise primarily from the physicochemical properties of singlet oxygen in water. Singlet oxygen has a much shorter lifetime in water than it does in organic solvents, causing challenges with respect to quantitative detection of singlet oxygen.The ensuing pages are an attempt to explore the theory and document the procedures developed to provide the accurate measurement of singlet oxygen in aqueous media. Details of this experimental method and singlet oxygen quantum yield results of new compounds relative to established photosensitizers will be presented.