Orientador: Ana Cláudia Pavarina / Banca: Carlos Eduardo Vergani / Banca: Vanderlei Salvador Bagnato / Resumo: O surgimento da resistência antifúngica aos tratamentos convencionais tem proporcionado o desenvolvimento de novas modalidades terapêuticas para o tratamento da candidose bucal. Nesse contexto, a utilização da PDT vem sendo sugerida como método alternativo para a inativação de microrganismos patogênicos. Este estudo avaliou a efetividade da PDT na inativação de C. albicans e C. glabrata, ATCC e resistente a fluconazol, por meio da utilização do e da iluminação com LEDs de agente fotossensibilizador Photogem® comprimento de onda azul. Inicialmente, os microrganismos avaliados foram inoculados em tubos de ensaio contendo meio de cultura líquido e incubados overnight a 37ºC. Em seguida, foram obtidas suspensões celulares das espécies de Candida avaliadas. Essas suspensões foram transferidas para placas de 96 orifícios, tratadas com cinco diferentes concentrações de Photogem® (2,5; 5; 10; 25 e 50mg/L) e expostas a quatro doses de luz (10,5; 18; 25,5 e 37,5J/cmø). Suspensões adicionais foram tratadas somente com as cinco concentrações do fotossensibilizador ou apenas com as quatro doses de luz. Cada condição experimental foi realizada três vezes. Após a realização desses experimentos, foram obtidas diluições seriadas de cada amostra (10-1 a 10-3 ), e alíquotas de 25 æL dessas diluições foram plaqueadas, em triplicatas, em Sabouraud Dextrose Agar. Adicionalmente, alíquotas de 25 æL foram removidas das cavidades das placas de orifícios e transferidas diretamente para um quadrante da placa de Petri, sem a realização de diluição. As placas foram incubadas a 37ºC por 48 horas. Após a incubação, foi realizada a contagem das colônias viáveis (ufc/mL), e os valores obtidos foram analisados com o teste t de Student (p < 0,05). (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Oral candidosis is an opportunistic infection that affects a significant percentage of the population. The oral infection caused by Candida spp. is usually treated with topical and systemic antifungal drugs. However, the widespread use of these agents has resulted in an alarming increase in the rate of antifungal resistance. Recently, the photodynamic therapy (PDT) has been studied as an alternative modality of killing microorganisms, including viruses, fungi and bacteria. The aim of this study was to determine whether Candida albicans and C. glabrata, ATCC and fluconazole-resistant strains, could be photosensitized by Photogem® in combination with blue Light Emitted Diode (LED). Suspensions of each Candida strain, containing 106 viable cells per milliliter, were treated with five concentrations of Photogem® (2.5, 5, 10, 25 and 50 mg/l), followed by LED irradiation in four light doses (10.5, 18, 25.5 and 37.5 J/cm2). Each experimental condition was carried out in triplicate and repeated tree times. From each sample, serial dilutions were obtained and aliquots of 25 æl of each dilution were plated on Sabouraud Dextrose Agar. All plates were incubated at 37°C for 48 hours. After incubation, colonies were counted (CFU/ml) and the data were statistically analyzed by the Student's t test (p < 0.05). The results demonstrated a concentration/dose-dependent pattern of inactivation, that resulted in complete elimination of all Candida evaluated. The minimal light dose for the complete inactivation of both C. albicans source was 18 J/cmø, in conjunction with 50 mg/l of Photogem®. After 25.5 and 37.5 J/cmø, a lower concentration of Photogem® was required to totally inactivate C. albicans ATCC (5 and 2.5 mg/l) in comparison with C. albicans luconazole-resistant (10 and 5 mg/l). (Complete abstract, clik electronic access below) / Mestre

Made available in DSpace on 2017-03-14T14:10:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-07-29. Added 1 bitstream(s) on 2017-03-14T14:42:45Z : No. of bitstreams: 1 000874567_20180729.pdf: 739207 bytes, checksum: 096d06efa6c645b9cd46c526ab46691f (MD5) Bitstreams deleted on 2018-07-31T11:21:36Z: 000874567_20180729.pdf,. Added 1 bitstream(s) on 2018-07-31T11:22:29Z : No. of bitstreams: 1 000874567.pdf: 2611072 bytes, checksum: c96b34ee605e5658c74f4eafc173b3c1 (MD5) / O presente estudo avaliou a eficácia da terapia fotodinâmica (PDT) na inativação de Candida albicans em modelos murino de candidíase oral. Adicionalmente, este estudo avaliou se a PDT causa danos ao núcleo da célula fúngica. Dessa forma, foram conduzidos três estudos: 1. Avaliou a eficácia da cloro-alumínio-ftalocianina encapsulado em nanoemulsão catiônica (ClAlP NE) na inativação de Candida albicans presente em modelo murino de candidíase oral; 2. Avaliou a eficácia da PDT mediada pelo Photodithazine®(PDZ) como tratamento para candidíase oral induzida; 3. Avaliou o potencial da inativação fotodinâmica (PDT) mediado pelo fotossensibilizador (PS) Curcumina (CUR) associado ao LED azul em causar danos no DNA de C albicans. No estudo 1, 75 camundongos fêmeas com 6 semanas foram imunossuprimidos e inoculados com C. albicans para induzir candidíase oral. A PDT foi realizada sobre a língua pela aplicação dos fotossensibilizadores ClAlP NE ou ClAlP DMSO e luz LED (100 J/cm2 - 660 nm). Em seguida, a avaliação microbiológica foi realizada por meio da recuperação de C. albicans da língua dos animais. Diluições seriadas foram realizadas, as colônias viáveis foram quantificado e o número de UFC mL determinado. Os animais foram sacrificados 24 horas após a PDT e as línguas foram removidas cirurgicamente para análise histológica e para análise de expressão de citocinas pró e antiinflamatórias (IL-1, TNF-α e IL-6) por ensaio de RT-qPCR. Os dados foram analisados por One-Way ANOVA. A PDT mediada por ClAIP-NE reduziu 2,26 log10 da viabilidade celular de C. albicans em comparação ao grupo controle (p <0,05). A PDT mediada por ClAlP DMSO não apresentou atividade antifúngica. A análise histológica revelou que a PDT não promoveu efeitos adversos sobre o tecido língual dos animais. A terapia mediada por ambos os fotossensibilizadores induziu a expressão... (Resumo completo, clicar acesso eletrônico abaixo) / This study evaluated the efficacy of photodynamic therapy (PDT) in the inactivation of C. albicans in a murine models of oral candidosis. Additionally, this study evaluated the poetencial of PDT to promote DNA damage of fungal cell. For this purpose, three studies were conducted: 1. Evaluated the photodynamic effect of chloro-aluminum phthalocyanine encapsulated in cationic nanoemulsions (ClAlP NE) to inactivate Candida albicans in a murine model of oral candidiasis. 2. Evaluated the photoinactivation (PDT) of Candida albicans in a murine model of oral candidosis, mediated by Photodithazine® (PDZ). 3. Evaluated the potential of PDT mediated by Curcumin (CUR) associated with the blue LED in causing DNA damage of C. albicans. In the first study, 75 6-week-old female Swiss mice were immunosuppressed and inoculated with C. albicans to induce oral candidiasis. PDT was performed on the tongue by the application of the photosensitizers and LED light (100 J cm-² - 660 nm). Next, microbiological evaluation was carried out by recovering C. albicans from the tongue via colony forming units (CFU mL-1). Then, the animals were sacrificed and the tongues were surgically removed for histological analysis and analysis of inflammatory cytokines expression (IL-1, TNF-α and IL-6) by RT-qPCR. Data were analyzed by ANOVA test. ClAlP NE-mediated PDT was able to reduce C. albicans recovered from the tongue (2.26 log10) when compared with control (p<0.05). ClAlP DMSO-mediated PDT did not present antifungal activity. Histological analysis revealed that PDT did not promote adverse effects on the tongue tissue. PDT mediated by both photosensitizers induced the TNF-α expression when compared with control (P<0.05). ClAlP mediated by PDT induced the expression of inflammatory cytokines, TNF-α, irrespective of the drug system delivery used. In the second study, a new murine model of oral...(Complete abstract electronic access below)

Made available in DSpace on 2014-06-11T19:33:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-12-03Bitstream added on 2014-06-13T18:45:12Z : No. of bitstreams: 1 almeida_jm_dr_araca.pdf: 895420 bytes, checksum: 5e46165cde5411fa955922cec8421c68 (MD5) / O objetivo do presente estudo foi avaliar histológica e histometricamente a influência da terapia fotodinâmica (PDT) no tratamento coadjuvante da doença periodontal (DP) experimental induzida em ratos diabéticos e não-diabéticos. Foram utilizados 180 ratos divididos em 2 grupos: Grupo A: 90 animais não-diabéticos e Grupo B: 90 animais diabéticos induzidos pela aloxana a 2% (42 mg/Kg). Nos Grupos A e B a DP foi induzida no primeiro molar inferior. Após 7 dias de evolução da DP, em todos animais de ambos os Grupos, a ligadura foi removida seguida de raspagem e alisamento radicular (RAR) e divididos conforme os tratamentos: I (RAR) - RAR e irrigação com 1 ml de soro fisiológico; II (TBO) - RAR e irrigação com 1 ml de azul de toluidina O (TBO 100 μg/ml); III (PDT) - RAR, irrigação com 1 ml de TBO e após 1 minuto irradiação com o laser em baixa intensidade. O laser (AsGaAl, 660 nm) foi aplicado na vestibular e lingual em 3 pontos eqüidistantes em cada região, com dose energética de 4 J/ponto. Dez animais de cada grupo experimental e tratamento foram sacrificados nos períodos de 7, 15 e 30 dias para a análise histológica e histométrica. Os dados histométricos foram submetidos à análise estatística (p<0,05). Os animais tratados com a PDT (Grupo A e B), apresentaram histologicamente tecido conjuntivo (TC) bem organizado e trabéculas ósseas presentes em toda a extensão da furca. Nos animais tratados com RAR nos dois grupos, o TC estava desorganizado, com intenso infiltrado inflamatório e áreas de reabsorções ósseas e cementárias. As alterações mais severas foram observadas no Grupo B. Histometricamente, no Grupo A, o tratamento com a PDT (0,33±0,05 mm2; 0,35±0,06 mm2; 0,27±0,07 mm2), promoveu menor perda óssea (p<0,05) na região de furca quando comparado com os animais tratados... / The aim of this study was to evaluate histological and histometrically the influence of photodynamic therapy (PDT) as an adjuvant treatment on the experimentally induced periodontitis in diabetic and non-diabetic rats. One-hundred eighty rats were assigned to 2 groups: Group A: Ninety non-diabetic and Group B: Ninety alloxan-diabetic. In Group B, after 7 days of alloxan-diabetic induction (42 mg/Kg), the periodontal disease (PD) was induced at the first mandibular molar. Seven days after the establishment of PD, the ligatures were removed. Thereafter, all animals were submitted to scaling and root planing (SRP) and then divided according with the following protocols treatment: I (SRP) – SRP and irrigation with saline solution (1 mL); II (TBO) – SRP and irrigation with 1 mL of Toluidine Blue (TBO) (100 μg/mL); III (PDT) – SRP, irrigation with 1mL of TBO and after 1 minute, low-level laser therapy (LLLT). The LLLT GaAlAs (4 J) was performed in 3 equidistant points at the each buccal and lingual aspect of the first mandibular molar. Ten animals in each experimental group and treatment were sacrificed at 7, 15 and 30 days. Histologic and histometric analyses were performed. The histometric values were statistical analyzed ( p<0.05). The animals treated by PDT in Group A and B showed the furcation area fulfilled with a well-organized connective tissue (CT) and with bone trabeculae in all extension of furcation. In animals treated with SRP, the CT was poorly organized, showing an intense inflammatory infiltrate and areas with bone and cement resorption. The worst tissue response was found in Group B. Histometrically, in Group A, the animals treated by PDT (0.33±0.05 mm2; 0.35±0.06 mm2; 0.27±0.07 mm2) showed a lesser bone loss (p<0.05) in all experimental... (Complete abstract click electronic access below)

Made available in DSpace on 2015-05-14T16:53:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-04-29Bitstream added on 2015-05-14T16:58:44Z : No. of bitstreams: 1 000821856.pdf: 381272 bytes, checksum: def92d15fe2de7528a92101a30b132f1 (MD5) / Introdução: A terapia fotodinâmica (TFD) se baseia em um conjunto de processos biológicos, físicos e químicos que ocorre por meio da ativação de um fotossensibilizador (FS) com luz (laser ou LED) para destruir as células alvo. O objetivo deste estudo foi avaliar a citotoxicidade e a resposta tecidual da TFD incluindo a produção de citocinas (IL - 1β e IL - 6). Métodos: 1) Para o teste de citotoxicidade os grupos foram divididos em: hipoclorito de sódio 5%; hipoclorito de sódio 2,5%; clorexidina 2%; solução salina; TFD (curcumina + luz Led azul); e controle (meio de cultura). As soluções foram diluídas em meio de cultura DMEM (1x104 células) e colocadas em placas de 24 poços, com linhagem de fibroblastos de camundongos (L-929). Depois de 6, 24 e 48 horas, o ensaio de MTT foi utilizado para avaliar a viabilidade celular e o ensaio de ELISA foi utilizado para avaliação de citocinas no sobrenadante. 2) Para o teste de resposta tecidual, os grupos foram divididos em: controle (solução salina); hipoclorito de sódio 2,5%; hipoclorito de sódio 5%; clorexidina 2%; TFD (curcumina + luz Led Azul). As soluções foram colocadas em tubos de polietileno e foram implantadas no tecido conjuntivo dorsal de ratos Wistar por 7, 15, 30, 60, e 90 dias. Os tubos com tecido circundante foram removidos e divididos ao meio e uma das metades foi corada com hematoxilina e eosina e a outra metade utilizada para avaliação das citocinas por ELISA. A análise estatística foi realizada utilizando ANOVA, Kruskal Wallis ou teste de Tukey (p < 0,05). Resultados: Quanto ao estudo in vitro, a TFD e a solução salina apresentaram baixa citotoxicidade semelhante ao do grupo controle (p> 0,05), menor que o hipoclorito de sódio (2,5% e 5%) e a clorexidina 2% (p < 0,05) em todos os períodos de tempo. Todas as soluções liberaram citocinas em quantidade similar... / Introduction: Photodynamic therapy (PDT) is based on a set of biological, physical and chemical processes that occurs through the activation of a photosensitizer (PS) with light (laser or LED) to destroy target cells. The aim of this study was to evaluate the cytotoxicity and tissue response of PDT including the production of cytokines (IL - 1β and IL - 6). Methods: 1) For the cytotoxicity test groups were divided into: 5% sodium hypochlorite; 2.5% sodium hypochlorite; 2% chlorhexidine; saline; PDT (curcumin + Led blue light); and control (culture medium). The solutions were diluted in DMEM culture medium (1x104 cells) and placed in 24 -well plates with mouse fibroblast line (L929). After 6, 24 and 48 hours, the MTT assay was used to assess cell viability and ELISA assay was used to assess cytokine in the supernatant. 2) To test the tissue response, the groups were divided into: control (saline); 2.5%; sodium hypochlorite, 5% sodium hypochlorite; 2% chlorhexidine; TFD (curcumin + Led blue light). The solutions were placed into polyethylene tubes and implanted in the dorsal tissue of Wistar rats for 7, 15, 30, 60, and 90 days. The tubes were removed with surrounding tissue and divided in half and one half was stained with hematoxylin and eosin and the other half used to assess cytokine by ELISA. Statistical analysis was performed using ANOVA, Kruskal Wallis or Tukey test (p < 0.05). Results: For the in vitro study, PDT and saline showed low cytotoxicity similar to the control group (p > 0.05) , less than sodium hypochlorite (2.5% and 5%) and 2% chlorhexidine (p < 0.05) at all time periods. All solutions released cytokines similar to control (p > 0.05) amount. As for the in vivo study, all solutions caused severe inflammatory reaction after 7 days, which decreased with time. No inflammation was observed at 90 days in the PDT group similar to the control (p > 0.05) . All materials...

Made available in DSpace on 2015-07-13T12:10:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-10. Added 1 bitstream(s) on 2015-07-13T12:25:45Z : No. of bitstreams: 1 000838217.pdf: 2068210 bytes, checksum: 7cc196b4afd9c4a009cb398bc82c6339 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O câncer de colo uterino (ou câncer cervical) é uma neoplasia maligna que tem a infecção pelo HPV como o principal fator de risco para o seu surgimento. As terapias atuais para o tratamento do câncer cervical têm como maior desvantagem a excisão substancial do estroma cervical e muitos efeitos adversos associados aos quimioterápicos, principalmente a cisplatina. Nesse cenário, a terapia fotodinâmica (TFD) vem se mostrando uma alternativa promissora, especialmente se combinada a tratamentos convencionais. A técnica é baseada na administração de um agente fotossensibilizador não tóxico ao paciente com posterior exposição a uma fonte de luz de comprimento de onda específico, resultando em resposta citotóxica via dano oxidativo. Diante disso, o presente estudo teve como objetivo avaliar, in vitro, a citotoxicidade, o tipo de morte induzido e os efeitos genotóxicos e mutagênicos da TFD em monoterapia e associada à cisplatina sobre as linhagens SiHa, C-33 A e HaCaT, utilizando os fotossensibilizadores Photogem e azul de metileno. Observou-se que tanto a terapia fotodinâmica mediada por azul de metileno (AMTFD) quanto a terapia mediada pelo Photogem (PGTFD) foram eficazes em reduzir o número de células viáveis, com a citotoxicidade sendo dependente da dose de luz. A cisplatina teve menor eficácia sobre as três linhagens quando comparada à TFD. Porém, a combinação cisplatina + TFD apresentou efeito sinérgico intenso e provocou maior morte celular em todas as condições testadas. A sequência de aplicação dos tratamentos (cisplatina + TFD ou TFD + cisplatina) demonstrou-se importante e dependente do fotossensibilizador. Com relação ao tipo de morte celular provocado, tanto a AMTFD quanto a PGTFD induziram morte independente de caspases, mas a AMTFD induziu morfologia típica de necrose, enquanto a PGTFD induziu morfologia mais similar à apoptose. A cisplatina induziu predominantemente apoptose e as terapias... / Cervical cancer is a malignant tumor that has HPV infection as the main risk factor for its emergence. Current therapies for the treatment of cervical cancer have as major disadvantage substantial excision of cervical stroma and many adverse effects associated with chemotherapy, especially cisplatin. In this scenario, photodynamic therapy (PDT) has proven to be a promising alternative, specially if combined with conventional treatments. The technique is based on the administration of a non-toxic photosensitizing agent to the patient with subsequent exposure to a light source of a specific wavelength, resulting in cytotoxic response by oxidative damage. Thus, the present study intended to evaluate in vitro cytotoxicity, the type of induced death and the genotoxic and mutagenic effects of PDT alone and associated with cisplatin on the cell lines SiHa, C-33 A and HaCaT, using the photosensitizers Photogem and methylene blue. It was observed that both the photodynamic therapy mediated by methylene blue (MBPDT) and Photogem mediated therapy (PGPDT) were effective in reducing cell viability with cytotoxicity being dependent on the light dose. Cisplatin was less effective over the three lines compared to PDT. However, the combination cisplatin + PDT had a greater synergistic effect and caused greater cell death in all conditions tested. The sequence of treatment application (PDT + cisplatin or cisplatin + PDT) was important and depended on the photosensitizer. Regarding the type of induced cell death, both AMPDT and PGPDT induced caspase-independent death, but AMPDT induced the typical morphology of necrosis, while PGPDT induced morphology was most similar to apoptosis. Cisplatin predominantly induced apoptosis; and combination therapy induced greater apoptosis- or necrosis-like rate according to the cell line, but always with a higher percentage of dead cells than monotherapies. MBPDT, either as monotherapy or in combination with cisplatin was ... / FAPESP: 12/25414-5

Made available in DSpace on 2015-08-20T17:10:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-04-14. Added 1 bitstream(s) on 2015-08-20T17:26:57Z : No. of bitstreams: 1 000840266_20160429.pdf: 112142 bytes, checksum: f753980f523bb8e5e7108026940280ab (MD5) Bitstreams deleted on 2016-05-04T13:08:27Z: 000840266_20160429.pdf,. Added 1 bitstream(s) on 2016-05-04T13:09:31Z : No. of bitstreams: 1 000840266.pdf: 2388303 bytes, checksum: 0cd278104ebd428e40cc96127a077555 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A ocorrência de uma variedade de agentes patogênicos resistentes aos antibióticos atuais continua a ser um problema, especialmente quando as infecções bacterianas estão crescendo em biofilme. Neste estudo, propomos o uso da terapia fotodinâmica (TFD) como monoterapia e também associada à antibioticoterapia como um tratamento alternativo. O objetivo deste estudo foi analisar os efeitos da TFD mediada pelo azul de metileno (AM) em Staphylococcus aureus (ATCC 25923) e Escherichia coli (ATCC 25922), como biofilme e em fase planctônica. Diferentes concentrações de fotossensibilizador (400; 200; 100; 50; 25; 12,5; 6,25 μg/mL) e fluência (2,8; 5,6; 11.2 22,4J/cm²) foram testadas. Também realizamos experimentos que avaliaram o efeito sinérgico da terapia fotodinâmica e o antibiótico ciprofloxacina. Os efeitos bactericidas da TFD como monoterapia não foram estatisticamente aumentados com a concentração de fotossensibilizador, e bactérias em biofilmes foram menos afetadas que as da fase planctônicas. Entretanto, o efeito sinérgico da terapia fotodinâmica seguida de ciprofloxacina no biofilme aumentou a redução bacteriana. Em biofilme para S. aureus, utilizando 50 ug/mL de AM, irradiação 22,4 J/cm² e a concentração de 15.625 ug/mL de antibiótico, tivemos uma redução de 61,80%. Para E. coli, houve uma erradicação bacteriana em todas as concentrações testadas na TFD combinada com antibiótico, tanto a irradiação de 11,2 J/cm², como a irradiação 22,4 J/cm². O crescimento bacteriano foi observado apenas na concentração mais baixa do antibiótico testada, de 0,125 ug/mL, em concentrações de 50 e 25 ug/mL de AM. / The occurrence of a variety of pathogens resistant to current antibiotics remains the major problem especially when bacterial infections are growing in biofilm. In this study, we propose the use of photodynamic therapy (PDT) as monotherapy and also associated with antibiotic therapy as an alternative treatment. The aim of this study was to analyze the effects of PDT mediated by methylene blue (MB) on Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) as biofilm and planktonic phase. Different concentrations of photosensitizer (400; 200; 100; 50; 25; 12,5; 6,25g/mL), and fluency of 2.8; 5.6; 11.2 22,4J/cm2 were tested. We also carried out experiments that evaluated the synergistic effect of photodynamic therapy and antibiotic ciprofloxacin. The bactericidal effects with PDT as monotherapy were not statistically increased with the concentration of photosensitizer and bacteria in biofilms were less than affect in the planktonic phase. Although, the synergistic effect of photodynamic therapy on biofilms followed by ciprofloxacin amplified the bacterial reduction. In biofilm for S. aureus, using 50 μg/ml MB, irradiation 22,4 J/cm² and the concentration of 15,625 μg/mL of antibiotic, got a reduction of 61,80%. For E. coli, there was a bacterial eradication at all concentrations tested in PDT combined with antibiotic, both the irradiation 11,2 J/cm² as the irradiation 22,4 J/cm². Bacterial growth was observed only in the lowest tested concentration of the antibiotic, 0,125 μg/mL, at concentrations of 50 and 25 μg/mL MB.

Made available in DSpace on 2014-08-13T14:51:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-02-20Bitstream added on 2014-08-13T17:59:39Z : No. of bitstreams: 1 000776228.pdf: 5343697 bytes, checksum: 6c774fdc58dfbb1de52eb34f163ddc1b (MD5) / O câncer de pele é o tipo mais frequente de câncer no Brasil, correspondendo a 25% de todos os cânceres registrados no País. Queratoses actínicas (QAs) são lesões pré-malignas comuns, com uma incidência estimada em 60% dos pacientes com idade acima de 60 anos, as QAs podem evoluir para carcinoma espinocelular (CEC) que é um tipo de câncer de pele não melanoma agressivo, podendo ser responsável por 34% das mortes por câncer de pele em todo mundo. A terapia fotodinâmica (TFD) em QAs com o ácido 5- aminolevulínico (ALA) ou aminolevulinato de metila (Metil-ALA) é uma opção de tratamento aceita para esta doença, devido à sua eficácia e excelentes resultados estéticos. O objetivo deste trabalho foi Avaliar e comparar a dor referida pelos pacientes, durante o tratamento de TFD utilizando o medicamento ALA e Metil-ALA a15%. Foram selecionados 40 pacientes com QAs em membros superiores, e realizado a divisão de dois grupos de 20 pacientes, grupo I administrado o medicamento ALA e no grupo II administrado Metil-ALA, em seguida os membros foram ocluídos permanecendo por 3 horas, após o curativo foi removido e realizado a iluminação da área por 40 minutos, com fluência de 36 Jzcm, no protótipo Kerato PDT a base de emissores de LED. Durante a iluminação e a cada 5 minuto foi aplicada uma escala visual numérica (EVN) para mensurar a dor referida. As lesões foram quantificadas e fotografadas antes e 30 dias após o tratamento. Os valores obtidos foram submetidos a análise estatística pelo teste t de Student, teste t pareado, teste de Friedman e teste de Tukey. Os resultados mostraram que não houve diferença estatisticamente significativa entre os dois grupos, e que ambos os medicamentos utilizados mostraram-se eficazes no tratamento de QAs. Comparando a dor referida pelos pacientes durante o tratamento, não houve diferença estatística entre o sexo feminino e o masculino, e nem entre os dois grupos, porém para o ... / Skin cancer is the most common type of cancer in Brazil, accounting for 25 % of all cancers registered in actinic keratoses country (AKs) are common precancerous lesions , with an estimated 60% of patients over the age incidence 60 years, AKs can progress to squamous cell carcinoma (SCC ) is a type of aggressive skin cancer melanoma, may be responsible for 34 % of deaths from skin cancer in the world. Photodynamic therapy (PDT) for AK with 5 - aminolevulinic acid ( ALA ) or methyl aminolevulinate ( Methyl- ALA ) is an accepted treatment option for this disease , due to its effectiveness and excellent cosmetic results . The aim of this study was to evaluate and compare the pain reported by the patients during the treatment of PDT using the medicine Methyl- ALA and ALA a15 %. AK 40 patients were selected for the upper limbs, the division performed and two groups of 20 patients, the drug administered group I and group II ALA administered -ALA methyl , then the remaining members were occluded for 3 hours, after curative was removed and held the lighting in the area for 40 minutes with a fluence of 36 J / em 2 , the prototype Kerato PDT -based LED emitters . During the lighting and every 5 minutes a numerical visual scale (NVS) was applied to measure referred pain . The lesions were imaged and quantified before and 30 days after treatment. Data were statistically analyzed by Student's paired t-test, Friedman test and Tukey 's t testo The results showed no statistically significant difference between the two groups, and that both drugs used were effective in the treatment of AKs . Comparing the pain reported by the patients during treatment , there was no statistical difference between the female and the male , nor between the two groups, although for males the pain scale was statistically higher when used Methyl- ALA . We conclude that both drugs may be indicated for the treatment of AKs scattered in the upper limb, and more studies focused ...

PART A The syntheses, spectroscopic and electrochemical characterisation of a series of titanium and tantalum phthalocyanine complexes are reported. The complexes are unsubstituted and substituted at either the peripheral or non-peripheral positions with sulphonates, aryloxy, arylthio or amino groups. The complexes mostly exhibit Qbands in the near-infrared region as well as interesting properties in different solvents. The interaction of differently sulphonated titanium and tantalum phthalocyanine complexes with methyl viologen (MV[superscript 2+]), and hence the stoichiometry and association constants are evaluated. Detailed photophysicochemical properties of the complexes were investigated and are for the first time presented with fluorescence lifetimes easily obtained from fluorescence quenching studies. The transformation of 1-hexene photocatalysed by aryloxy- and arylthio-appended complexes is also presented for the first time. The electrochemical properties of the complexes are unknown and are thus presented. Cyclic (CV) and square wave (SWV) voltammetries, chronocoulometry and spectroelectrochemistry are employed in the study of the complexes. Two oneelectron reductions and a simultaneous 4-electron reduction are observed for the unsubstituted Cl[subscript 3]TaPc. Reduction occurs first at the metal followed by ring-based processes. The tetra- and octa-substituted derivatives however exhibit peculiar electrochemical behaviour where a multi-electron transfer process occurs for complexes bearing certain substituents. For all complexes, the first two reductions are metal-based followed by ring-based processes. A comparative study of the electrocatalytic activities of the complexes towards the oxidation of nitrite is also investigated. The complexes are immobilised onto a glassy carbon electrode either by drop-dry or electropolymerisation methods. All the modified electrodes exhibit improved electrocatalytic oxidation of nitrite than the unmodified electrodes by a twoelectron mechanism producing nitrate ions. Catalytic currents are enhanced and nitrite overpotential reduced to ~ 0.60 V. Kinetic parameters are determined for all complexes and a mechanism is proposed. PART B: The syntheses and electrochemical characterisation of chromium and titanium complexes for the selective trimerisation of ethylene to 1-hexene are presented. The synthesis of the chromium complex requires simple steps while tedious steps are used for the air-sensitive titanium complex. The spectroscopic interaction of the chromium complex with the co-catalyst methylaluminoxane is investigated. The complexes are characterised by electrochemical methods such as cyclic voltammetry and spectroelectrochemistry.

The synthesis and extensive spectroscopical characterization of novel phthalocyanines are reported. The new compounds were characterized by elemental analysis, FT-IR, ¹HNMR, mass spectrometry and UV–Vis spectroscopy. The new phthalocyanines showed remarkable photophysicochemical behaviour. The novel phthalocyanines were then conjugated to nanoparticles, silver and ZnO. The coupling of the novel Pcs to nanoparticles was through covalent bonding and ligand exchange. These conjugates were supported onto electrospun polystyrene fibers and chitosan microbeads for use as photocatalysts. The efficiency of the immobilized Pcs and Pc-nanoparticles was assessed by the phototrasfromation of organic pollutants, methyl orange and Rhodamine 6G as model dyes. Upon conjugating phthalocyanines to nanoparticles, there was a great increase in the rate of photodegradation of the model dyes. The photodynamic activity of the novel phthalocyanines upon conjugating to nanoparticles and selected targeting agents is also reported. The targeting agents employed in this study are folic acid and polylysine. Conjugating the phthalocyanines to folic acid or polylysine improved the solubility of the phthalocyanines in aqueous media. The potency of the conjugates was investigated on breast (MCF-7), prostate and melanoma cancer cell lines. The phthalocyanines showed no toxicity in the absence of light. However, upon illumination, a concentration dependent cellular decrease was observed.

Orientador: Ana Cláudia Pavarina / Banca: Juliana Campos Junqueira / Banca: Gilberto Leite Braga Ubida / Banca: Lívia Nordi Dovigo / Banca: Marlise Inêz Klein Furlan / Resumo: O presente estudo avaliou a eficácia da terapia fotodinâmica (PDT) na inativação de Candida albicans em modelos murino de candidíase oral. Adicionalmente, este estudo avaliou se a PDT causa danos ao núcleo da célula fúngica. Dessa forma, foram conduzidos três estudos: 1. Avaliou a eficácia da cloro-alumínio-ftalocianina encapsulado em nanoemulsão catiônica (ClAlP NE) na inativação de Candida albicans presente em modelo murino de candidíase oral; 2. Avaliou a eficácia da PDT mediada pelo Photodithazine®(PDZ) como tratamento para candidíase oral induzida; 3. Avaliou o potencial da inativação fotodinâmica (PDT) mediado pelo fotossensibilizador (PS) Curcumina (CUR) associado ao LED azul em causar danos no DNA de C albicans. No estudo 1, 75 camundongos fêmeas com 6 semanas foram imunossuprimidos e inoculados com C. albicans para induzir candidíase oral. A PDT foi realizada sobre a língua pela aplicação dos fotossensibilizadores ClAlP NE ou ClAlP DMSO e luz LED (100 J/cm2 - 660 nm). Em seguida, a avaliação microbiológica foi realizada por meio da recuperação de C. albicans da língua dos animais. Diluições seriadas foram realizadas, as colônias viáveis foram quantificado e o número de UFC mL determinado. Os animais foram sacrificados 24 horas após a PDT e as línguas foram removidas cirurgicamente para análise histológica e para análise de expressão de citocinas pró e antiinflamatórias (IL-1, TNF-α e IL-6) por ensaio de RT-qPCR. Os dados foram analisados por One-Way ANOVA. A PDT mediada por ClAIP-NE reduziu 2,26 log10 da viabilidade celular de C. albicans em comparação ao grupo controle (p <0,05). A PDT mediada por ClAlP DMSO não apresentou atividade antifúngica. A análise histológica revelou que a PDT não promoveu efeitos adversos sobre o tecido língual dos animais. A terapia mediada por ambos os fotossensibilizadores induziu a expressão... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study evaluated the efficacy of photodynamic therapy (PDT) in the inactivation of C. albicans in a murine models of oral candidosis. Additionally, this study evaluated the poetencial of PDT to promote DNA damage of fungal cell. For this purpose, three studies were conducted: 1. Evaluated the photodynamic effect of chloro-aluminum phthalocyanine encapsulated in cationic nanoemulsions (ClAlP NE) to inactivate Candida albicans in a murine model of oral candidiasis. 2. Evaluated the photoinactivation (PDT) of Candida albicans in a murine model of oral candidosis, mediated by Photodithazine® (PDZ). 3. Evaluated the potential of PDT mediated by Curcumin (CUR) associated with the blue LED in causing DNA damage of C. albicans. In the first study, 75 6-week-old female Swiss mice were immunosuppressed and inoculated with C. albicans to induce oral candidiasis. PDT was performed on the tongue by the application of the photosensitizers and LED light (100 J cm-² - 660 nm). Next, microbiological evaluation was carried out by recovering C. albicans from the tongue via colony forming units (CFU mL-1). Then, the animals were sacrificed and the tongues were surgically removed for histological analysis and analysis of inflammatory cytokines expression (IL-1, TNF-α and IL-6) by RT-qPCR. Data were analyzed by ANOVA test. ClAlP NE-mediated PDT was able to reduce C. albicans recovered from the tongue (2.26 log10) when compared with control (p<0.05). ClAlP DMSO-mediated PDT did not present antifungal activity. Histological analysis revealed that PDT did not promote adverse effects on the tongue tissue. PDT mediated by both photosensitizers induced the TNF-α expression when compared with control (P<0.05). ClAlP mediated by PDT induced the expression of inflammatory cytokines, TNF-α, irrespective of the drug system delivery used. In the second study, a new murine model of oral...(Complete abstract electronic access below) / Doutor