Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2018-11-16T19:33:03Z No. of bitstreams: 1 ClaudiaSofiadeFreitasCorreiaPopovTeseParcial2017.pdf: 709320 bytes, checksum: 74c3fb3e34fa9828bc490124e9d19b7b (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2018-11-16T19:33:23Z (GMT) No. of bitstreams: 1 ClaudiaSofiadeFreitasCorreiaPopovTeseParcial2017.pdf: 709320 bytes, checksum: 74c3fb3e34fa9828bc490124e9d19b7b (MD5) / Made available in DSpace on 2018-11-16T19:33:23Z (GMT). No. of bitstreams: 1 ClaudiaSofiadeFreitasCorreiaPopovTeseParcial2017.pdf: 709320 bytes, checksum: 74c3fb3e34fa9828bc490124e9d19b7b (MD5) Previous issue date: 2018-12-13 / Anurans represent a valuable source of bioactive molecules. These substances constitute a primary line of defense against predators and pathogenic microorganisms such as bacteria and fungi. Secreted peptides by granular glands represent a source of potential pharmacological agents with diverse functions. The structural and functional characterization of these biomolecules allows a better understanding of their function and biotechnological applications. Thus, in this work, the new peptides were isolated from H. raniceps with variable size between 11 and 16 amino acid residues, all of which contain two cysteines in their primary structure and with immunomodulatory activity: AC12, DK16 and RC11. A nuclear magnetic resonance analysis allowed to determinate a structure of these β-sheet molecules, as well as the presence of disulfide bridges that gives stability to the peptides. Regarding the structural analysis, none of the samples showed effective antimicrobial action. The immunomodulatory action profile of the samples was also checked. For this, a cytotoxicity essay of the molecules was initially tested. AC12 and DK16 did not reduce the viability of RAW 264.7 cells, decreasing inflammatory markers such as NO, IL-12 and TNF-α. They also decreased IL-10 production, a pro-inflammatory cytokine, but associated with several types of neoplasms. The peptide RC11 was shown to be toxic in RAW 264.7 strain. However, all samples were shown to be safe when tested at different concentrations in erythrocytes of Balb/c mice, presenting low hemolysis rates. In the in vivo tests, the three peptides reduced the paw edema caused by carrageenan, standing out the peptide AC12. Such data were corroborated by histological analysis of paw tissue. Regarding the immunomodulatory action with carrageenan-induced peritonitis assay, peptide AC12 was again the most significant, reducing the TNF-α dosed from the peritoneal lavage and increasing IL-10. In conclusion, AC12 has shown to be a good candidate as an anti-inflammatory agent, and could be used in the treatment of several inflammatory conditions. / Os anuros representam uma fonte valiosa de moléculas bioativas. Essas substâncias constituem a linha de defesa primária contra predadores e microrganismos patogênicos como bactérias e fungos. Peptídeos secretados pelas glândulas granulares representam uma fonte de potenciais agentes farmacológicos com funções diversas. A caracterização estrutural e funcional dessas biomoléculas permite um melhor entendimento da sua função e aplicações biotecnológicas. Assim, neste trabalho foram descritos três novos peptídeos isolados de H. raniceps com tamanho variável entre 11 e 16 resíduos de aminoácidos, todos eles contendo duas cisteínas na sua estrutura primária e com propriedades imunomoduladoras: AC12, DK16 e RC11. A análise por ressonância magnética nuclear permitiu determinar a estrutura dessas moléculas em folha-β, bem como a presença de pontes de dissulfeto que fornecem resistência e estabilidade às moléculas. Referente à análise estrutural, nenhuma das amostras mostrou ter ação antimicrobiana efetiva. Foi também verificado o perfil de ação imunomoduladora das amostras. Para tal, inicialmente foi testada a citotoxicidade das moléculas. AC12 e DK16 não alteraram a viabilidade das células RAW 264.7, diminuindo marcadores inflamatórios como NO, IL-12 e TNF-α. Também diminuíram a produção de IL-10, uma citocina pró-inflamatória, porém, associada a vários tipos de neoplasias. O peptídeo RC11 mostrou ser tóxico na linhagem RAW 264.7. Porém, todas as amostras mostraram-se seguras quando testadas em diferentes concentrações em eritrócitos de camundongos Balb/c, apresentando taxas de hemólise pouco significativas. Nos testes in vivo, os três peptídeos reduziram o edema da pata provocado por carragenina, sobressaindo o peptídeo AC12. Tais dados foram corroborados pela análise histológica do tecido da pata. Relativamente à ação imunomoduladora no ensaio de peritonite provocada por carragenina, novamente, o peptídeo AC12 teve destaque, reduzindo o TNF-α dosado do lavado peritoneal e aumentando a IL-10. Deste modo, o peptídeo AC12 mostrou ser um bom candidato a agente anti-inflamatório, podendo ser utilizado no tratamento de diversos quadros inflamatórios.

O óleo de copaíba é um produto natural encontrado principalmente na região amazônica, onde é utilizado na medicina popular como tratamento para inflamações e como cicatrizante. A espécie Copaifera multijuga Hayne demonstrou um potencial efeito anti-inflamatório em relação a outras espécies de Copaífera L., tendo como principal responsável o composto majoritário β-cariofileno. Nosso grupo de pesquisa vem estudando a veiculação deste óleo em nanoemulsões e desenvolveu uma formulação que contém uma elevada proporção de óleo de copaíba no núcleo oleoso (20 % w/v), sem prejuízo da estabilidade do sistema. A partir desta formulação, seguiram-se os estudos de avaliação da permeação cutânea, de otimização da formulação e de avaliação de atividade anti-inflamatória, apresentados neste trabalho. Um método em cromatógrafo a gás acoplado à espectrômetro de massas no modo headspace (HS-CG/EM) foi validado a fim de analisar β-cariofileno em mostras de pele provenientes do teste de permeação cutânea com nanoemulsões de óleo de copaíba. O método mostrou-se específico, linear, preciso e exato. O teste de permeação cutânea demonstrou que apenas com a nanoemulsão é possível detectar β-cariofileno na camada da derme, enquanto que, com o óleo, somente no estrato córneo. A seguir, foram testados dois tensoativos catiônicos na formulação da nanoemulsão para verificar se a carga positiva na interface da gotícula poderia promover a permeação cutânea do β-cariofileno. O tensoativo brometo de cetiltrimetilamônio provou ser mais eficiente em concentrações consideradas seguras para o uso tópico, revertendo o potencial zeta para valores adequados, sem interferir no tamanho de gotícula e índice de polidispersão. O tensoativo oleilamina também reverteu o potencial zeta, porém somente em concentrações muito elevadas, podendo ser consideradas tóxicas. O teste de permeação cutânea demonstrou que a incorporação de tensoativos catiônicos aumenta a retenção de β-cariofileno na epiderme em três vezes, enquanto que, na derme, não há diferença estatística entre as formulações aniônica e catiônica. Após, as nanoemulsões escolhidas foram incorporadas em hidrogéis de Carbopol®, Natrosol® e quitosana com vistas ao espessamento e adequação da viscosidade ao uso tópico. O hidrogel com quitosana 8 apresentou-se instável, com aumento de tamanho de gotícula e índice de polidispersão, apesar de não ter afetado o potencial zeta. Os hidrogéis de Carbopol® e Natrosol® apresentaram bons resultados de caracterização físico-química, porém somente o hidrogel de Natrosol® foi escolhido para os estudos de permeação cutânea e atividade anti-inflamatória in vivo, devido ao seu caráter neutro. No estudo de permeação cutânea das nanoemulsões incorporadas em hidrogel, houve um aumento na retenção de β-cariofileno na derme em relação às nanoemulsões, e houve a facilitação de permeação até o fluido receptor. Por fim, o estudo da atividade in vivo das formulações selecionadas demonstrou que o óleo de copaíba apresenta atividade anti-inflamatória e que a sua incorporação em nanoemulsões aumenta este efeito. No entanto, ambas nanoemulsões, tanto negativamente quanto positivamente carregadas, apresentaram resultado semelhante para a inibição do edema. Quando compara-se a permeação cutânea das formulações verifica-se que na derme não há diferença estatística, o que pode justificar a semelhança do grau de inibição da inflamação no teste in vivo. Em relação à atividade das nanoemulsões incorporadas em hidrogéis, pode-se verificar que no teste de edema de pata apenas a formulação carregada positivamente teve um efeito mais pronunciado, enquanto no edema de orelha as formulações obtiveram um perfil equivalente ao controle cetoprofeno, porém não potencializaram o efeito do óleo. Quanto ao estudo com os parasitos causadores da Leishmaniose, os testes in vitro mostraram que os tratamentos (óleo de copaíba, β-cariofileno e suas nanoemulsões) foram mais eficazes contra as espécies L. major e L. donovani em comparação às espécies L. amazonensis e L. braziliensis. O teste in vivo mostrou que todos os tratamentos foram capazes de reduzir a área da ferida dos camundongos infectados com L. major. No entanto, eles não conseguiram recuperar totalmente os animais. / Copaiba oil is a natural product found especially in the Amazon region, where it is used as treatment for inflammations and as wound healer in the popular medicin. The species Copaifera multijuga Hayne showed a potential anti-inflammatory effect in relation to other Copaifera L. species, mainly due to its major compound, β-caryophyllene. Our research group studied the oil incorporation in nanoemulsions and developed a formulation containing a high copaiba oil proportion in the oil core (20% w/v) without loss of system stability. Based in this nanoemulsion, we present in this study the skin permeation, the formulation optimization and the anti-inflammatory activity. A method in gas chromatograph coupled with mass spectrometer in headspace mode (HS-GC/MS) was validated to analyze β-caryophyllene in skin samples from the skin permeation assay. The method proved to be specific, linear, precise and accurate. Skin permeation test showed that only with the nanoemulsion is possible to detect β-caryophyllene in the dermis layer, while with the oil, only in the stratum corneum. After, two cationic surfactants were tested in the nanoemulsion to prove if the positive charge on the droplet interface could promote β-caryophyllene permeation. Cetyltrimethylammonium bromide proved to be more effective at concentrations considered safe for topical use, reversing the zeta potential to suitable values, without interfering with droplet size and polydispersity index. Oleylamine also reversed the zeta potential, but only at very high concentrations, that may be considered toxic. Skin permeation test showed that the incorporation of cationic surfactants increases β-caryophyllene retention in the epidermis by three fold, whereas in the dermis, there is no statistical difference between the cationic and anionic nanoemulsions. Afterward, the chosen nanoemulsions were incorporated in Carbopol®, Natrosol® and chitosan hydrogels in order to adjust the viscosity for topical use. Chitosan hydrogel presented instability, with an increase in droplet size and polydispersity index, although it has not affected the zeta potential. Carbopol® and Natrosol® hydrogels showed good results in physicochemical characterization, but only Natrosol® hydrogel was chosen for the following skin permeation 10 and anti-inflammatory activity in vivo studies due to its neutral character. In nanoemulsions thickened-hydrogel skin permeation study, there was an increase in the β-caryophyllene retention in the dermis compared to nanoemulsions, and promoted permeation to the receptor fluid. Finally, in vivo anti-inflammatory activity from selected formulations showed that the copaiba oil has anti-inflammatory activity and that its incorporation into nanoemulsions increases this effect. However, both negative and positively charged nanoemulsions, showed similar results for inhibition of edema. When the nanoemulsions’ skin permeation is compared, it is found that in the dermis there is no statistical difference, which may explain the similarity degree of inflammation inhibition in the in vivo test. Regarding to the activity of the nanoemulsions incorporated in hydrogels, it can be verified that in paw edema assay, only the positively charged formulation had a more pronounced effect, whereas in the ear edema the formulations obtained a profile equivalent to the control, ketoprofen, but did not potentiate the effect of the oil. As for the study with the parasites causing Leishmaniasis, in vitro tests showed that the treatments (copaiba oil, β-caryophyllene and their nanoemulsions) were more effective against L. major and L. donovani compared to L. amazonensis and L. braziliensis. In vivo assay showed that all treatments were able to reduce the wound area of mice infected with L. major. However, they were unable to fully recover the animals from the disease.

Ho Toi Sze Joyce. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 65-74). / Abstracts in English and Chinese. / Acknowledgement --- p.ii / Contents --- p.iii / Abstract --- p.viii / List of Abbreviations --- p.xvii / List of Tables --- p.xix / List of Figures --- p.xx / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- The role of Non-steroidal anti-inflammatory drugs (NSAIDs) --- p.1 / Chapter 1.2 --- NSAID-induced gastrointestinal (GI) toxicity --- p.1 / Chapter 1.2.1 --- Pathogenesis of NSAID-induced GI toxicity --- p.2 / Chapter 1.2.2 --- GI symptoms --- p.4 / Chapter 1.2.3 --- GI ulcers --- p.4 / Chapter 1.2.4 --- GI complications --- p.5 / Chapter 1.2.5 --- Risk factor for GI complications --- p.6 / Chapter 1.2.6 --- Ulcerogenicity of different NSAIDs in upper GI events --- p.6 / Chapter 1.3 --- Prevention of NSAID-induced GI toxicity --- p.7 / Chapter 1.3.1 --- H2-receptor antagonists --- p.8 / Chapter 1.3.2 --- Misoprostol --- p.8 / Chapter 1.3.3 --- Proton Pump Inhibitor (PPI) --- p.9 / Chapter 1.3.4 --- Selective COX-2 Inhibitors --- p.10 / Chapter 1.3.4.1 --- GI safety of selective COX-2 inhibitors --- p.11 / Chapter 1.3.4.1.1 --- Gastrointestinal outcomes research of rofecoxib --- p.13 / Chapter 1.3.4.1.2 --- Celecoxib Long term Arthritis Safety Study --- p.14 / Chapter 1.3.4.2 --- Cardiovascular toxicity of NSAIDs --- p.15 / Chapter 1.3.4.2.1 --- Cardiovascular toxicity of non-selective NSAIDs --- p.15 / Chapter 1.3.4.2.2 --- Cardiovascular toxicity of selective COX-2 inhibitors --- p.16 / Chapter 1.4 --- Guidelines on the management of osteoarthritis (OA) and rheumatoid arthritis (RA) --- p.21 / Chapter 1.4.1 --- American College of Rheumatology (ACR) Subcommittee --- p.22 / Chapter 1.4.2 --- National Institute for Clinical Excellence (NICE) --- p.23 / Chapter 1.4.3 --- Hong Kong Hospital Authority (HA) --- p.23 / Chapter 1.5 --- Cost of illness of upper GI events in the setting of an emergency room of a regional hospital in Hong Kong and cost analysis of selective COX-2 inhibitor with non-selective NSAID plus gastroprotective agent --- p.24 / Chapter 1.6 --- Objectives --- p.25 / Chapter Chapter 2 --- Cost of illness of upper GI events in the setting of an emergency room of a regional hospital in Hong Kong --- p.26 / Chapter 2.1 --- Methods --- p.28 / Chapter 2.1.1 --- Study site --- p.28 / Chapter 2.1.2 --- Cohort participants --- p.28 / Chapter 2.1.3 --- Resource data collection --- p.29 / Chapter 2.1.4 --- Cost data --- p.30 / Chapter 2.1.5 --- Statistical Methods --- p.31 / Chapter 2.1.6 --- Study perspective --- p.31 / Chapter 2.2 --- Results --- p.31 / Chapter 2.2.1 --- Demographic data --- p.31 / Chapter 2.2.2 --- Total direct medical cost of upper GI complaints in UCH --- p.33 / Chapter 2.3 --- Discussion --- p.35 / Chapter 2.3.1 --- Total direct medical cost of upper GI events --- p.35 / Chapter 2.3.2 --- Cost of upper GI events associated with NSAID usage --- p.38 / Chapter 2.3.3 --- Low dose aspirin on NSAID-induced GI toxicity --- p.38 / Chapter 2.3.4 --- Limitation --- p.39 / Chapter 2.3.5 --- Future study --- p.41 / Chapter 2.4 --- Conclusion --- p.41 / Chapter Chapter 3 --- Cost analysis of selective COX-2 inhibitor versus non-selective NSAID with gastroprotective agent --- p.43 / Chapter 3.1 --- Methods --- p.46 / Chapter 3.1.1 --- Local randomized clinical trial --- p.46 / Chapter 3.1.1.1 --- Study population --- p.46 / Chapter 3.1.1.2 --- Cost data --- p.47 / Chapter 3.1.1.3 --- Statistical Methods --- p.48 / Chapter 3.1.1.4 --- Sensitivity analysis --- p.49 / Chapter 3.1.2 --- Large randomized clinical trial --- p.49 / Chapter 3.1.2.1 --- Study population --- p.49 / Chapter 3.1.2.2 --- Cost data --- p.50 / Chapter 3.2 --- Results --- p.50 / Chapter 3.2.1 --- Local randomized clinical trial --- p.51 / Chapter 3.2.1.1 --- Demographic data --- p.51 / Chapter 3.2.1.2 --- Cost analysis --- p.52 / Chapter 3.2.1.3 --- Sensitivity analysis --- p.53 / Chapter 3.2.2 --- Large randomized clinical trial --- p.54 / Chapter 3.2.2.1 --- Demographic data --- p.54 / Chapter 3.2.2.2 --- Cost analysis --- p.55 / Chapter 3.3 --- Discussion --- p.55 / Chapter 3.3.1 --- Cost analysis --- p.55 / Chapter 3.3.2 --- Sensitivity analysis --- p.59 / Chapter 3.3.3 --- Low dose aspirin on NSAID-induced GI toxicity --- p.59 / Chapter 3.3.4 --- Limitation --- p.60 / Chapter 3.4 --- Future study --- p.62 / Chapter 3.5 --- Conclusion --- p.62 / Chapter Chapter 4 --- Conclusion --- p.63 / Chapter Chapter 5 --- Reference --- p.65 / Appendix Data collection form --- p.75

Celecoxib is a COX-2 selective non-steroidal anti-inflammatory drug which has been shown to inhibit growth and induce apoptosis in various cancer cell lines. Using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and an apoptosis detection kit, we demonstrated that celecoxib was able to induce growth inhibition and apoptosis in a dose-dependent manner in 3 NPC cell lines: HK-1, Hone-1, and C666-1. Afterwards, a proteomic approach was used to study the underlying mechanisms involved in celecoxib-mediated effects on two COX-2 positive NPC cell lines (HK-1 and C666-1). Results showed that a total of 18 protein spots were differentially expressed in the HK-1 and C666-1 cells. On the other hand, we also compared the proteomic expression profile between an NPC cell line (C666-1) and a normal nasopharynx cell line (NP69) in order to study whether those differentially expressed proteins after celecoxib treatment were also involved in NPC carcinogenesis. Proteomics results with confirmation using Western blotting discovered that HSP27 phosphorylated of serine 82 (HSP27-pSer82) protein was up-regulated in C666-1 cells when compared with that in NP69 cells. After treatment with celecoxib, expression of HSP27-pSer82 protein was down-regulated in both HK-1 and C666-1 cells. These findings suggest that down-regulation of HSP27-pSer82 protein expression may have mediated the growth-inhibitory effects of celecoxib in HK-1 and C666-1 cells. Finally, other differential expressed proteins identified from proteomics with confirmation by immunocytochemical staining in the 2 NPC cell lines and 40 NPC patient specimens showed that down-regulation of annexin 2 and beta2-tubulin may be important in NPC formation. / COX-2 over-expression has been found in various cancers such as colorectal cancer, liver cancer and lung cancer. In vivo studies have shown that mice overexpressing COX-2 developed breast cancer whereas COX-2 knockout mice had reduced rates of cancer formation in the intestines and skin. In the present study, COX-2 expression in NPC patient biopsies was examined and correlated with the clinicopathological data of the patients. Immunocytochemical staining showed that COX-2 protein was over-expressed in 84.6% (66/78) of non-metastatic NPC patients and was associated with an advanced nodal stage (P<0.05). All these data support an important role for COX-2 in NPC pathogenesis. / In summary, this study is the first to identify HSP27-pSer82 protein as a potential target of celecoxib in NPC cells. Detailed investigations of the functional role of molecular targets identified in this study would improve our understanding of the chemotherapeutic effects of celecoxib and, in the long run, may lead to a more effective chemotherapeutic treatment to this common cancer. / Nasopharyngeal carcinoma (NPC) is prevalent in southern China. Although early stage patients have a high rate of cure with radiotherapy alone, the prognosis for those with stage III or IV disease remains poor due to subsequent development of distant metastases. Therefore there is an urgent need to develop novel biologic agents to improve treatment outcomes. / Chan, Ming Lok. / Adviser: Anthony T.C. Chan. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3418. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 141-171). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

De nos jours, de nombreuses maladies chroniques telles que le cancer ou l’arthrose nécessitent encore de nouvelles modalités de traitement. Des biomatériaux naturels capables de véhiculer des substances actives font partie des solutions à cette problématique. Récemment, des travaux ont été menés sur un nouveau type de biomatériau, les Complexes de Polyélectrolytes Compacts (CoPEC). Dans le cadre de cette thèse, des CoPEC à base de polyélectrolytes biosourcés, le chitosan et l’alginate, fonctionnalisés avec la β-cyclodextrine (βCD) ont été formulés. Le CoPEC βCD-chitosan/alginate, non-cytotoxique, a présenté des propriétés anti-inflammatoires intrinsèques dans le cadre d’un modèle in vitro d’inflammation. De plus, ce CoPEC a présenté une capacité à contenir et relarguer deux substances actives hydrophobes modèles, le piroxicam et la prednisolone. Enfin, une stratégie d’inclusion de substances actives hydrophiles au sein du matériau a été mise en œuvre. Le nouveau CoPEC est prometteur car il peut exposer un effet anti-inflammatoire intrinsèque et d’autres effets thérapeutiques via l’inclusion de substances actives au sein des cyclodextrines. / Nowadays, many chronic diseases, such as cancer or osteoarthritis, still need new modalities of treatment. Natural biomaterials able to convey active substances represent a solution to this problematic. Lately, several research works have been conducted on a new type of biomaterial named Compact Polyelectrolyte Complexes (CoPEC). As part of this thesis, CoPEC have been prepared from two biosourced polyelectrolytes, chitosan and alginate, functionalized with β-cyclodextrin (βCD). Through an in vitro inflammation model, the non-cytotoxic βCD-chitosan/alginate CoPEC has displayed intrinsic anti-inflammatory properties. Moreover, this CoPEC has demonstrated a capacity to host and release piroxicam and prednisolone, two model hydrophobic active substances. Finally, a strategy to include hydrophilic active substances into the material has been implemented.Thus, the newly CoPEC is promising because it can exhibit an intrinsic anti-inflammatory effect as well as other therapeutic effects through the inclusion of active substances into the cyclodextrins.

Resumo: Os metabólitos do AA exercem um reconhecido papel na patogênese da doença periodontal. O objetivo deste trabalho foi avaliar o efeito do celecoxib, um inibidor seletivo da enzima cicloxigenase-2 (COX-2), sobre o desenvolvimento de doença periodontal induzida por ligadura em ratos. Após a colocação de ligadura de algodão ao redor dos primeiros molares inferiores direitos, 180 ratos Holtzman foram aleatoriamente subdivididos em 3 grupos experimentais com 60 animais cada, os quais receberam diariamente dose oral de celecoxib 10 mg ou 20 mg/ kg de peso corporal (grupos Ce1 e Ce2, respectivamente) ou, dose oral de 10ml/kg de NaCl a 0,9% (grupo Controle). Aos 3, 5, 10, 18 e 30 dias após o início do experimento, 12 animais de cada grupo experimental foram sacrificados. O tratamento com celecoxib, em ambas as concentrações, reduziu significantemente (p<0.05) a perda óssea alveolar radiográfica aos 5 dias e, diminuiu a intensidade da reabsorção óssea, observada histologicamente, aos 30 dias. Ainda, o celecoxib atrasou o início e, diminui a magnitude, do processo inflamatório agudo. Estes resultados demonstram que a inibição seletiva da COX-2 com o celecoxib, pode interferir com a resposta do tecido periodontal frente à presença de ligadura em ratos. / Abstract: Arachidonic acid metabolites have a recognized role in the pathogenesis of periodontal disease. The purpose of this study was to evaluate the effect of a selective cyclooxygenase-2 (COX-2) inhibitor, celecoxib, on the progression of periodontal disease in a ligature-induced periodontitis model in rats. After ligature placement in the mandibular right first molars, 180, 6-week-old Holtzman rats were ramdomly assigned to one of the following groups of treatment that consisted in a daily oral dose of 10mg/kg body weight of celecoxib (Ce1), 20mg/kg body weight of celecoxib (Ce2) or 10ml/kg of 0,9%NaCl (Control). At 3, 5, 10, 18 and 30 days later, 12 animals of each group were sacrificed. Treatment with celecoxib significantly (p < 0.05) decreased the radiographic bone loss at 5 days of experiment and, decreased the bone loss activity, histologically observed at 30 days. In addition, celecoxib was shown to delay the onset and to suppress the magnitude of the acute inflammatory process. These results show that selective cyclooxygenase-2 (COX-2) inhibition with celecoxib, can interfer with the periodontal tissue response to ligature placement in rats. / Orientador: Luís Carlos Spolidorio / Coorientador: Elcio Marcantonio Junior / Banca: Joni Augusto Cirelli / Banca: Maria Angela Naval Machado / Mestre

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2016-03-02T17:03:56Z No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) PDF - Airlla Laana de Medeiros Cavalcanti.pdf: 2086060 bytes, checksum: b74e6eb69ebe6e0583efc0e2d23bedac (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2016-06-13T20:33:03Z (GMT) No. of bitstreams: 2 PDF - Airlla Laana de Medeiros Cavalcanti.pdf: 2086060 bytes, checksum: b74e6eb69ebe6e0583efc0e2d23bedac (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-06-13T20:33:10Z (GMT). No. of bitstreams: 2 PDF - Airlla Laana de Medeiros Cavalcanti.pdf: 2086060 bytes, checksum: b74e6eb69ebe6e0583efc0e2d23bedac (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-02-13 / Some inflammatory skin diseases are treated with drugs carried on conventional dosage forms that often fail to achieve adequate concentrations in the tissue to generate maximum pharmacological effect. Pentoxifylline (PTX) is one of these drugs widely studied due to its anti-inflammatory activity by inhibiting the production of TNFα and other proinflammatory cytokines. The use of PTX incorporated into microemulsions (ME) would be a novel alternative for the treatment of inflammatory skin disorders. This new drug delivery system can be used topically and can be able to increase the permeation through skin and the effectiveness of several drugs compared to conventional treatments. The aim of this work was to develop a microemulsion containing PTX (PTX-ME) for topical use. The formulation obtained from the pseudoternary phase diagrams was characterized and evaluated using methods such as polarized light microscopy (MLP), differential scanning calorimetry (DSC) and transmission electron microscopy (TEM). The in vitro release profile was determined using the model of Franz cells and antiedematogenic activity in vivo was determined using the technique of paw edema induced by carrageenan. The ME was composed of distilled water (5%), acid–capric/caprylic triglycerides (51%), Tween 80 (39.6%) and Brij TM 52 (4.4%). Analyzes of MLP, DSC, and TEM were able to confirm that the ME obtained was water-in-oil (W/O) type. The formulation was thermodynamically stable against thermic stress. Beside this, it had physicalchemical characteristics that allow its topical use. In vitro release of PTX-ME followed the Higuchi kinetic model. Additionally, it showed significant anti-inflammatory activity in paw edema induced by carrageenan in all stages of the assay. Consequently, the PTX-ME showed an interesting alternative for treatment of dermatological disorders. / Algumas doenças dermatológicas inflamatórias são tratadas com fármacos veiculados em formas farmacêuticas convencionais, que muitas vezes não atingem concentrações teciduais adequadas para gerar o efeito farmacológico máximo. A pentoxifilina (PTX) é um desses fármacos e tem sido amplamente estudada com relação a sua atividade anti-inflamatória por inibir a produção de TNF-α e outras citocinas pró-inflamatórias. O uso da PTX incorporada a uma microemulsão (ME) seria uma alternativa inédita para o tratamento de afecções dermatológicas inflamatórias. Esse novo sistema transportador de fármaco pode ser utilizado topicamente e é capaz de aumentar a permeação cutânea e a eficácia de vários fármacos em relação aos tratamentos convencionais. O objetivo deste trabalho foi desenvolver um sistema microemulsionado contendo PTX para aplicação tópica. A formulação obtida a partir de um diagrama de fase pseudoternário foi caracterizada e avaliada utilizando métodos como microscopia de luz polarizada (MLP), calorimetria exploratória diferencial (DSC) e microscopia eletrônica de transmissão (MET). O perfil de liberação in vitro foi determinado utilizando o modelo de células de Franz e a atividade antiedematogênica in vivo foi determinada através da técnica de edema de pata induzido por carragenina. A ME desenvolvida foi constituída por 5% de água destilada, 51% de triglicerídeos do ácido cáprico e caprílico, 39,6% de Tween ® 80 e 4,4% Brij ® 52. Através das análises de MLP, DSC e MET foi possível confirmar a estruturação do sistema como ME do tipo água em óleo (A/O). A formulação apresentou-se estável frente a estresses térmicos, além de possuir características físico-químicas que possibilitam seu uso por via tópica. A liberação in vitro da ME-PTX obedeceu ao modelo cinético de Higuchi e apresentou atividade anti-inflamatória significativa em edema de pata induzido por carragenina em todas as etapas do ensaio. Portanto, pôde-se concluir, que a veiculação da PTX através de um sistema microemulsionado mostrou-se uma alternativa interessante e inédita para o tratamento de afecções dermatológicas.

Made available in DSpace on 2015-04-11T13:38:40Z (GMT). No. of bitstreams: 1 Dissertacao Final Alcineide Magalhaes 2010.pdf: 1214565 bytes, checksum: c15b666de091c06216476d80e7f8573f (MD5) Previous issue date: 2010-11-05 / Fundação de Amparo à Pesquisa do Estado do Amazonas / This study investigated the efficacy of Marsypianthes chamaedrys Vahl inflorescence and leaf extracts in inhibiting the inflammatory and coagulant actions of Bothrops atrox venom. M. chamaedrys, which is used in Brazil as a folk medicine to treat snakebites and local inflammatory reactions, was tested in vitro to determine its ability to block indirect phospholipase A2 and direct coagulant activities and in vivo to determine its ability to inhibit leukocyte migration, cytokine release and an increase in vascular permeability. In vitro, M. chamaedrys showed antiphospholipase A2 and anticoagulant activities; the latter activity was also confirmed by prothrombin time (PT) and activated partial thromboplastin time (aPTT) in the absence of venom. Of the extracts used, those obtained from the crushed plant had the greater inhibitory activity in in vitro tests, showing that biological activity is affected by the way extracts are obtained. In vivo, M. chamaedrys inhibited leukocyte migration and the release of the proinflammatory cytokines IL-6 and TNF- without altering the concentration of the anti-inflammatory cytokine IL-10; however, it did not inhibit an increase in vascular permeability. In preliminary tests, M. chamaedrys showed no acute toxicological or cytotoxic effects. As specific antivenoms are not effective in neutralizing the local action of Bothrops venoms, characterization of the anti-inflammatory mechanisms induced by M. chamaedrys is of vital importance if the extracts of this plant species are to be used in future as adjuvants in the treatment of snakebites. / Marsypianthes chamaedrys, espécie vegetal amplamente distribuída no Brasil, é utilizada popularmente em acidentes ofídicos e reações inflamatórias locais. No presente trabalho M.. chamaedrys foi testada frente às seguintes atividades do veneno de Bothrops atrox: inflamatória (fosfolipásica A2, migração de leucócitos, liberação de citocinas e aumento da permeabilidade vascular) e coagulante. In vitro, Marsypianthes chamaedrys foi eficaz em bloquear as atividades fosfolipásica A2 e coagulante, apresentando também atividade anticoagulante frente aos testes TAP e TTPA. Dos extratos obtidos, a forma de contuso apresentou melhor atividade inibitória nos ensaios in vitro, demonstrando que a atividade biológica é afetada pelo modo de obtenção dos extratos. In vivo, os extratos inibiram a migração de leucócitos e a liberação de citocinas pró-inflamatórias IL-6 e TNF-α, sem alterar significamente, a concentração da citocina anti-inflamatória IL-10; porém, não inibiram o aumento da permeabilidade vascular. Em ensaios preliminares, M. chamaedrys não demonstrou efeitos toxicológicos agudos ou efeitos citotóxicos. A caracterização dos mecanismos anti-inflamatórios induzidos por Marsypianthes chamaedrys é de extrema importância para que extratos obtidos dessa espécie vegetal possam, futuramente, ser utilizados como coadjuvante no tratamento, visto que os antivenenos específicos não são eficazes na neutralização das atividades locais, nos acidentes botrópicos.

Vários mecanismos foram propostos para reduzir a toxicidade gástrica dos fármacos antiinflamatórios. A complexação com zinco foi proposta considerando-se o fato de que o zinco possui ação protetora sobre a mucosa gástrica. O complexo piroxicam-zinco apresenta atividades antiinflamatória e analgésica semelhantes às do fármaco livre. No entanto, a modificação de propriedades físico-químicas acarretou alteração do perfil cinético do fármaco. A fim de investigar as diferenças farmacocinéticas , o complexo piroxicam-zinco foi estudado quanto à farmacocinética em sangue e linfa de ratos. Estudaram-se também os efeitos do complexo na mucosa gástrica em comparação com o piroxicam livre em modelos de úlcera induzida por ácido acetilsalicílico e por etanol. Estudos cinéticos foram conduzidos em ratos Wistar machos que receberam doses equivalentes de 10 mg/kg de piroxicam ou piroxicam-zinco por via oral através de sonda de polipropileno. O sangue e a linfa dos animais foram coletados em pequenos intervalos por período de 22 horas. O piroxicam foi extraído das amostras, utilizando-se mistura de cicloexano/éter e as determinações quantitativas feitas através de cromatografia líquida de alta eficiência. Além desses ensaios, testou-se a solubilidade do complexo e determinou-se o coeficiente de partição em noctanol-tampão fosfato utilizando o método de \"shake-flask\". Os resultados mostram que o perfil plasmático do complexo piroxicam-zinco é semelhante ao do fármaco livre tanto em sangue quanto em linfa, mostrando, no entanto, absorção mais lenta da forma complexada. Estes dados podem ser explicados pela menor solubilidade do complexo e pela diferença encontrada no coeficiente de partição de piroxicam e piroxicam-zinco. Nos modelos estudados, verificou-se que o piroxicam-zinco não apresenta vantagem sobre a forma livre, no que se refere ao agravamento de úlcera gástrica / Several procedures have been proposed for reducing the gastric toxicity of anti-inflammatory drugs. Among them, complexation with zinc has been recommended on the grounds that zinc exerts a protective action on the gastric mucosa. Indeed, zinc-piroxicam complex shows anti-inflammatory and analgesic activities similar to those provided by the free drug. Complexation, however, not only modifies the physico-chemicat properties of the free drug, but also alters its kinetic profile. In order to investigate such differences, the pharmacokinetics of zinc-piroxicam complex was studied in the blood and Iymph of rats. In addition, the effects of this drug complex on the gastric mucosa, as compared to those of free piroxicam, were studied in ASA- and ethanol-induced ulcer models, and through the determination of mucus in the gastric barrier. Kinetic studies were performed in mate Wistar rats receiving equivalent doses (10 mg/kg) of piroxicam or zinc-piroxicam orally by means of a polypropylene probe. Both blood and Iymph samples were collected at intervals during a period of 22 hours. Piroxicam was extracted from the samples employing a mixture of cyclohexane/ether, and determinations were performed through HPLC. Solubility of the drug complex was also assessed, and the partition coefficient was determined in n-octanolphosphate buffer employing the shake-flask method. Results showed the plasmatic profile of zinc-piroxicam complex to be similar to that of the free drug, both in blood and in Iymph, although absorption was slower in the complexed form These results can be explained by low solubility of the zinc-piroxicam and by the differences found for the partition coefficients of piroxicam and zinc-piroxicam. Overall, in the models employed, the use of zinc-piroxicam was not found to be advantageous over that of free piroxicam regarding the aggravation of gastric ulcers

Made available in DSpace on 2015-04-22T22:14:16Z (GMT). No. of bitstreams: 1 Fernanda Guilhon Simplicio.pdf: 4539844 bytes, checksum: 540712a8f10fe1279182d4c859c53c91 (MD5) Previous issue date: 2009-06-30 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The Byrsonima genus (Malpighiaceae) is widely used as a medicine in Brazil. The tea made of the stem bark of B. japurensis A. Juss., popularly know as ―sara-tudo‖, is indiscriminately used against many inflammatory disorders in Amazonas State, in spite of the fact that there is no scientific data to support this usage. Thus, in this work we performed a pharmacognostic characterization of the stem bark of this specie and a pharmacological study with an aqueous extract obtained from it. In the pharmacognostic study we applied pharmacopeic and no pharmacopeic methods to determinate quality parameters of the herbal drug. In the pharmacological study we evaluated its antioxidant, antiplatelet and antiedematogenic capacity, in order to investigate its anti-inflammatory potential and possible mechanisms. To evaluate the safety of usage of an aqueous extract with therapeutic purposes we also carried out an acute toxicity study. The pharmacognostic study of the stem bark of B. japurensis showed that this herbal drug has a variety of substances with pharmacological potential. This fact was observed in the pharmacological study, where the substances of extract acted by different mechanisms culminating in an appreciable anti-inflammatory activity. However, the same composition variety was responsible for the high toxicity presented by the extract. Nevertheless, the species proved to be promising as a source of prototypes for new medicines. However, the species proved to be promising, after further study of the dose-effect in vivo, for use in therapy, or even as a source of new prototypes of drugs / Espécies do gênero Byrsonima (Malpighiaceae) são amplamente empregadas como medicamento em todas as regiões do Brasil. O chá da casca do caule de Byrsonima japurensis A. Juss., conhecida popularmente como sara-tudo, é indiscriminadamente utilizado contra diversas doenças inflamatórias pela população do Estado do Amazonas. Porém, não havia estudos científicos que validassem esse emprego. Por esta razão, este trabalho fez uma caracterização farmacognóstica da casca do caule desta espécie e um estudo farmacológico de do extrato aquoso dessa droga vegetal. No estudo farmacognóstico foram utilizadas metodologias farmacopéicas e não farmacopéicas visando determinar alguns parâmetros de qualidade da droga vegetal. No estudo farmacológico foram realizadas avaliações da capacidade antioxidante, antiagregante plaquetária e antiedematogênica do extrato aquoso, visando investigar o potencial anti-inflamatório e possíveis mecanismos. Para avaliar a segurança do uso do extrato com finalidades terapêuticas, também foi realizada uma avaliação de toxicidade aguda. O estudo farmacognóstico da casca do caule de B. japurensis, entre outros resultados, mostrou que essa droga vegetal possui uma grande diversidade de substâncias com potencial farmacológico. Esse fato foi refletido no estudo farmacológico, onde as substâncias presentes no extrato atuaram por diferentes mecanismos culminando numa apreciável atividade anti-inflamatória. Entretanto, essa mesma diversidade parece ser responsável pela toxicidade relativamente alta apresentada pelo mesmo. Contudo, a espécie mostrou ser promissora, após mais estudos da relação dose-efeito in vivo, para aplicação na terapêutica, ou mesmo como fonte de novos protótipos de fármacos.