Abstarct Analysis of the Therapy in Patients with Juvenile Idiopathic Arthritis Author: Martina Procházková Tutor: PharmDr. Josef Malý, Ph.D. Department of Social and Clinical Pharmacy, Faculty of Pharmacy in Hradec Králové, Charles University Introduction and aims: Methotrexate (MTX) is an effective and safe drug used for the treatment of juvenile idiopathic arthritis (JIA) and is well-known like a golden standart in the treatment of this serious illness. The aim of this study was analysis of therapy in patients with JIA focusing on MTX and faktors affecting its intolerance. Methods: The study was undertaken from January 2017 to February 2018 at the Department of Paediatrics and Adolescent Medicine, 1st Faculty of Medicine, Charles University in Prague. Patients younger than 18 years old, who have become ill with JIA for the first time or repeatedly between years 2015-2017 and were treated by MTX were included in this study. The prevalence of MTX intolerance was measured by "Methotrexate Intolerance Severity Score" questionaire every 3 months during regular rheumatological examinations. Simultaneously, medical records were reviewed. Data analysis was performed by means of descriptive statistics, correlation dependence and statistical signifikance of dependence was assessed with statistical two-choice...

Rheumatoid arthritis (RA), the most common autoimmune disease, affects billions of people worldwide. Conventional therapeutics do not provide satisfactory efficacy and even cause severe adverse reactions. Researchers are seeking new approaches for RA management. Toll-like receptor 4 (TLR4) signalling plays a pivotal role in the pathogenesis of RA, and has been proposed as a potential therapeutic target for RA. Chinese medicines are believed to be alternative options for conventional RA therapeutics. A herbal formula RL, consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos, has traditionally been used in treating various inflammatory disorders including RA. In this study we assessed the anti-arthritic efficacy of RL in animals, and investigated the involvement of TLR4 signalling in RL's effects in vivo and in vitro. In vivo anti-arthritic efficacy of RL was evaluated using CIA (collagen-induced arthritis) rats, a model that is well established for studying human RA. Articular disease manifestations were investigated grossly, radiographically, and histologically. Isolated splenocytes were used to determine the effects of RL on immune responses. Molecular events in the TLR4 pathways upon RL treatment were examined in sera and joint tissues of CIA rats as well as in cultured lipopolysaccharide (LPS)-stimulated murine RAW264.7 and human THP-1 cells.

Made available in DSpace on 2015-07-22T20:49:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-04-27 / Objetivo: Determinar as concentrações séricas de 25-hidroxicolecalciferol (25(OH)D), cálcio sérico total, fósforo sérico, fosfatase alcalina e paratormônio (PTH) em pacientes com artrite idiopática juvenil (AIJ) poliarticular e associá-las com a atividade e duração da doença, com a densidade mineral óssea e com o uso de medicamentos. Método: Por meio de um estudo transversal e controlado, foram avaliados 30 pacientes com AIJ poliarticular, pareados por sexo e idade com 30 controles saudáveis. Realizou-se avaliação clínica, antropométrica e laboratorial nos pacientes e controles e densitometria óssea nos pacientes. Resultados: Dos 30 pacientes incluídos no estudo, 23 (76,7%) eram meninas, 16 (53,3%) não caucasóides e com média de idade de 14 anos (4 a 20 anos). O tempo médio de doença foi 5 anos. Concentrações médias de cálcio sérico total e fosfatase alcalina foram significantemente mais baixas nos pacientes com AIJ comparativamente aos controles (p< 0,0001 e p= 0,001, respectivamente). Em relação a 25(OH)D, PTH e fósforo sérico não se observaram diferenças entre os grupos AIJ e controle. Quanto às concentrações séricas de 25(OH)D, 8 pacientes (26,7%) e 5 controles (16,7%) apresentaram valores compatíveis com deficiência (inferiores a 20 ng/ml) e 14 pacientes (46,7%) e 18 controles (60%) apresentaram valores compatíveis com insuficiência (entre 20–32 ng/ml). Estes valores não se associaram com a atividade de doença, com o uso de medicamentos ou densidade mineral óssea. Conclusão: Observamos elevada frequência de insuficiência e deficiência de 25(OH)D nas amostras estudadas. O comprometimento do metabolismo ósseo enfatiza a importância do acompanhamento evolutivo dos pacientes com AIJ. / Objective: To determine serum concentrations of 25-hydroxycholecalciferol (25(OH)D) and biochemical markers of bone turnover in patients with polyarticular juvenile idiopathic arthritis (JIA) and to associate them with disease activity and duration, with bone mineral density and with use of medications. Methods: In a cross-sectional and controlled study, 30 patients with polyarticular JIA were assessed, compared with 30 age and sex-matched healthy subjects. We evaluated clinical, anthropometric, laboratory parameters and dual-energy x-ray absorptiometry (DEXA). Results: Out of 30 patients included in the study, 23 (76.7%) were girls, 16 (53.3%) no caucasian and the mean age was 14 years old (4 to 20 years). The mean time of disease was 5 years. The mean serum calcium and phosphatase alkaline were significantly lower in patients with JIA compared to controls (p<0.0001 and p=0.001, respectively). There was no difference between the serum concentrations of 25(OH)D, PTH and phosphorus between patients and controls. Based on serum 25(OH)D concentrations, 8 patients (26.7%) and 5 controls (16.7%) were vitamin D deficient (< 20 ng/ml) and 14 patients (46.7%) and 18 controls (60%) were vitamin D insufficient (between 20 and 32 ng/ml). These values were not associated with disease activity, use of medications or bone mineral density. Conclusions: A high frequency of 25(OH)D insufficiency and defficiency was observed in the study population. The involvement of biochemical bone markers emphasizes the importance of the bone metabolism follow-up in patients with JIA. / TEDE / BV UNIFESP: Teses e dissertações

A área das Ciências Farmacêuticas busca constantemente por tratamentos mais eficientes, direcionados para alvos específicos, com diminuição da dose necessária e com a minimização dos efeitos adversos. Neste contexto, a área de Nanotecnologia Farmacêutica apresenta grande potencial de aplicabilidade, com resultados bastante promissores para o tratamento de diversas doenças. Os sistemas nanoestruturados têm sido avaliados para a incorporação de fármacos já utilizados em tratamentos administrados formas farmacêuticas convencionais que apresentam problemas farmacocinéticos ou farmacodinâmicos quando administrados. E, também, para a incorporação de novas moléculas com potencial para o tratamento de determinada doença. Neste trabalho de tese, nanocápsulas de núcleo lipídico versáteis contendo metotrexato na forma ácida e éster, bromelina, etanercept e infliximab foram desenvolvidas buscando contornar as limitações e aumentar a eficácia terapêutica desses fármacos. Inicialmente, as propriedades anti-inflamatórias de nanocápsulas de núcleo lipídico revestidas por micelas de polissorbato 80 contendo metotrexato encapsulado foram avaliadas em experimentos in vitro e in vivo, em células mononucleares obtidas a partir do líquido sinovial de pacientes com artrite reumatoide e em ratos Lewis com artrite induzida por adjuvante completo de Freund, respectivamente. As nanocápsulas de núcleo lipídico demonstraram serem altamente eficazes no controle da inflamação, sendo que os efeitos anti-inflamatórios in vivo foram alcançados em doses 75% menores que o metotrexato em solução. Na sequência, o tratamento in vitro da linhagem de células de carcinoma de mama humano, MCF-7, com nanocápsulas de núcleo lipídico multiparede funcionalizadas com bromelina demonstrou uma redução de 160 vezes na concentração necessária para obter o mesmo efeito quando comparada a uma solução de bromelina. A influência das pseudofases aniônicas e catiônicas no mecanismo de distribuição da indometacina, tacrolimus, aciclovir, metotrexato e éster etílico de metotrexato, foram avaliadas aplicando um algoritmo desenvolvido para nanocápsulas de núcleo lipídico. Verificou-se que somente a indometacina sofreu influência da presença de cargas, aumentando a afinidade pela fase dispersa das formulações. Formulações de nanocápsulas de núcleo lipídico multiparede contendo metotrexato na forma ácida e éster encapsulados e/ou funcionalizando a superfície das nanocápsulas foram desenvolvidas e testadas in vitro em linhagens de células tumorais (MCF-7) e em linhagens de células sadias (HaCaT). Essas formulações demonstraram atividade antiproliferativa maior para as MCF-7 (com redução em mais de 50% na viabilidade celular) em comparação com as soluções de metotrexato e éster etílico de metotrexato e esta atividade foi maior para as formulações em que as moléculas foram funcionalizadas na superfície das nanopartículas. A captação das nanopartículas pelas células também foi maior para as formulações funcionalizadas com metotrexato ou éster etílico de metotrexato em comparação com a formulação em que o éster de metotrexato está encapsulado. As três formulações contendo metotrexato na forma ácida ou éster não demonstraram ação antiproliferativa em linhagens de células sadias (HaCaT). Devido à baixa expressão de receptores de folato nessas células, não houve aumento da captação celular em comparação à formulação sem fármaco. Por último, foram desenvolvidas satisfatoriamente formulações de nanocápsulas de núcleo lipídico multiparede funcionalizadas com os anticorpos monoclonais infliximab e etanercept, e contendo éster etílico de metotrexato encapsulado, demonstrando que são adequadas para futuros estudos visando o tratamento da artrite reumatoide. Esse conjunto de resultados demonstra que as nanocápsulas de núcleo lipídico com funcionalização de superfície versátil, sejam revestidas com polissorbato 80 ou multiparede funcionalizadas são um sistema bastante promissor para a administração de fármacos de modo a aumentar sua especificidade e eficácia. / The Pharmaceutical Sciences field is constantly searching for more effective treatments, aiming specific targets, with dose reduction and minimization of side effects. In this context, the Pharmaceutical Nanotechnology field presents great applicability potential, with highly promising results for the treatment of several diseases. Nanostructured systems have been evaluated for the encapsulation of drugs approved for use in conventional pharmaceutical dosage forms that, however, exhibit pharmacokinetic or pharmacodynamics problems when administered, and for the encapsulation of novel molecules with potential to treat a determined disease. In the present thesis, versatile lipid-core nanocapsules containing methotrexate in the acid and ester forms, bromelain, etanercept and infliximab were developed, seeking to circumvent the limitations and increase the therapeutic efficacy of these drugs. Initially, the anti-inflammatory properties of methotrexate-loaded lipid-core nanocapsules coated with polysorbate 80 micelles were evaluated in in vitro and in vivo experiments, using mononuclear cells obtained from the synovial fluid of rheumatoid arthritis patients and Lewis rats with Freund complete adjuvant-induced arthritis. Lipid-core nanocapsules demonstrated to be highly effective in the control of inflammation, and the in vivo anti-inflammatory effects were reached in a dose 75% lower than the methotrexate in solution. In the sequence, the in vitro treatment of a human breast cancer cell line, MCF-7, with bromelina-functionalized multiple-wall lipid-core nanocapsules demonstrated a 160-fold reduction of the concentration required to obtain the same effect when compared with a bromelain solution. The influence of the anionic and cationic pseudo-phases in the distribution mechanism of indomethacin, tacrolimus, acyclovir, methotrexate and methotrexate ethyl ester was evaluated through an algorithm developed for lipid-core nanocapsules. It was verified that only indomethacin underwent influence in the presence of charge, increasing the affinity by the disperse phase of the formulations. Multiple-wall lipid-core nanocapsules formulations containing methotrexate in the acid and ester forms encapsulated and/or functionalizing the surface of the nanoparticles were developed and tested in vitro in tumour MCF-7 cells and in a healthy cell line (HaCaT). These formulations demonstrated higher anti-proliferative activity for the MCF-7 cells (reduction of over 50 % in cellular viability) in comparison with the methotrexate and methotrexate ethyl ester solutions and this activity was higher for the formulations in which the molecules were functionalized in the surface of the nanoparticles. A higher cellular uptake was observed for the formulations functionalized with methotrexate or methotrexate ethyl ester in comparison with the formulations in which the methotrexate ester is encapsulated. The three formulations containing methotrexate in the acid or ester form did not demonstrate anti-proliferative activity in non-tumour cell lines (HaCaT). Since these cells have a small expression of folate receptors, the uptake was not increased in comparison with the formulation without drug. Lastly, formulations of methotrexate ethyl ester-loaded multiwall lipid core nanocapsules functionalized with monoclonal antibodies infliximab and etanercept were successfully developed demonstrating suitability for future studies aiming the treatment of rheumatoid arthritis. These groups of results demonstrate that versatile lipid core nanocapsules, either coated with polysorbate 80 or multiwalled functionalized are a very promising system for the administration of drugs aiming their specificity and efficacy.

INTRODUÇÃO Fibromialgia (FM) está presente em 20% dos pacientes com artrite reumatoide (AR), podendo superestimar escore de atividade de doença com 28 articulações (DAS28) e interferir no uso de medicamentos modificadores do curso da doença (MMCD). Escore ultrassonográfico com sete articulações (US7) para quantificação de proliferação/efusão sinovial por escala de cinza (US7-EC) e de vascularização sinovial por power Doppler (US7-PD) são medidas objetivas de sinovite. OBJETIVOS Avaliar o impacto da FM nos escores US7-EC e US7-PD em pacientes com AR. MÉTODOS Estudo de caso-controle pareado por duração da AR e positividade do fator reumatoide, aninhado a coorte de pacientes com AR. Pontuação de imagens feita de forma cegada. Coeficientes de concordância kappa intraobservador e interobservador com observador externo mais experiente e cegado foram calculados. Coeficientes de correlação de Spearman (rs) para DAS28, US7-EC e US7-PD entre grupos foram calculados. Valores de P<0,05 foram considerados estatisticamente significativos (bicaudal). RESULTADOS Setenta e duas mulheres com 57,5 (49,3-66,8) anos de idade, AR há 13 (6-19) anos e em uso de MMCD convencional (97%) e biológico (18%) foram incluídas: 36 com FM (casos) e 36 sem FM (controles). DAS28 foi maior entre casos: 5,2 (4,3-6,3) vs. 4,0 (3,3-4,6), P<0,001. US7-EC e US7-PD foram iguais entre casos e controles: 10,0 (7,0-11,0) vs. 9,0 (7,0-11,0), P=0,37 e 3,0 (1,0-5,8) vs. 4,0 (2,0-5,0), P=0,87, respectivamente. DAS28, US7-EC e US7-PD se correlacionaram em ambos os grupos (rs=0,35 a 0,39, P<0,05), exceto DAS28 e US7-PD na presença de FM (rs=0,12, P=0,50). CONCLUSÃO FM não impacta a avaliação de sinovite por US7-EC e US7-PD em pacientes com AR. US7-PD parece ser melhor do que US7-EC na identificação de pacientes com AR com DAS28 superestimado pela FM. Estudos longitudinais com alvos terapêuticos baseados em escores clínicos e ultrassonográficos permitirão comparar acurácia dos métodos, bem como conhecer os efeitos no valor do tratamento. / BACKGROUND Fibromyalgia (FM) is present in 20% of rheumatoid arthritis (RA) patients. FM may overestimate RA disease activity score with 28 joints (DAS28) and impact disease modifying antirheumatic drugs (DMARD) treatment decision. Gray-scale 7-joint ultrasound score (US7) for synovial proliferation/effusion (US7-GS) and power Dopper for synovial vascularization (US7-PD) are objective measures of synovitis. OBJECTIVE To study the impact of FM in US7-GS and US7-PD among RA patients. METHODS This is a case-control study matched by RA duration and rheumatoid factor positivity, nested by a cohort of RA patients. Image scoring was blinded. Intra- and inter-rater kappa agreement coefficients with an experienced blinded external assessor were calculated. Spearman´s rank test correlation coefficients (rs) among DAS28, US7-GS and US7-PD were calculated. P values<0.05 were considered statistically significant (two-tailed). RESULTS Seventy two women aged 57.5 (49.3-66.8) years, with RA for 13 (6-19) years, in use of conventional (97%) and biologic DMARD (18%) were included: 36 with FM (cases) and 36 without FM (controls). DAS28 was greater among cases: 5.2 (4.3-6.3) vs. 4.0 (3.3-4.6), P<0.001. US7-GS and US7-PD were similar among cases and controls: 10.0 (7.0-11.0) vs. 9.0 (7.0-11.0), P=0.37 and 3.0 (1.0-5.8) vs. 4.0 (2.0-5.0), P=0.87, respectively. DAS28, US7-GS and US7-PD correlated in both groups (rs=0.35 to 0.39, P<0.05), except for DAS28 and US7-PD in the presence of FM (rs=0.12, P=0.50). CONCLUSION FM does not impact synovitis appraisal by US7-GS and US7-PD in RA patients. US7-PD may be better than US7-GS to identify RA patients with DAS28 overestimated by FM. Longitudinal studies with clinical and ultrasound-based scores as treatment targets are necessary to compare both methods accuracy and their effects on treatment value.

Motoric system diseases belong to the most frequent ones in the present days. All age categories are affected. Congenital defects are included and also injuries, inflammations, degenerative illnesses and others. In the Czech republic are many centers gathering patients with the same diseases. However the centers for orthopedic patients are missing. The dissertation is divided into a theoretic and empiric parts. The theoretic one is focused generally on problems with joint replacements, rehabilitation and relation between a medic and a patient. The dissertation was targeted on: 1. To verify if the informing about the subsequent postop nursing care provided in the center for patients undergoing aloplasty comes up to a patient?s expectations and fulfils their needs. 2. To find out if hospital nurses perceive the difference in the patient engagement in the nursing care according to if he had visited the center before or not. 3. To reveal potential problems in the center running from the patient, nurse and doctor point of view. The research part of the dissertation was implemented with a combination of both qualitative and quantitative survey. For quantitative survey was chosen a method of questioning with a form technique for patients (after a hip or a knee aloplasty) in the Czech republic and Austria and also for nurses working in the orthopedic department and intensive care unit in the Pelhřimov hospital. Three hypotheses were established. 1. The given nursing care informations provided in the center for patients undergoing aloplasties fulfil the patient?s needs. 2. The hospital nurses assess a significant difference in the patient?s collaboration between those who had visited the center before and who had not. 3. In the center for patients waiting for aloplasty daily running there exist problems from the nurse?s point of view. As for the qualitative part of survey a method of questioning with the technique of a semistructured dialog with open questions was used. The research file was created by patients after principal joints aloplasties. In this part we have tried to answer three main questions. 1. What is the relationship between both hospital and the center nurse activities and the patient?s contentment in the sphere of his needs? 2. What sorts of informations for the patients in the center are missing? 3. The patients evaluate the informations provided in the center as orientational or as significant for easier postop care managing? The results of the research survey were analysed and taken in schemes. Patients suffer from disturbing their both physical and psychological needs and so the goal of collaboration is a satisfied and fully informed patient. From the results it is apparent that a nurse in the center closely cooperates with a hospital nurse. The survey revealed that the patients in the center are missing no informations. The patients are prepared for the operation, orient themselves in the related problems know what comes with the rehabilitation as an important part of the postop care. To be concluded: patients are prepared for easier postop care managing. On the basis of analysis of respondent?s answers it was found that the patients undergoing a principal joint aloplasty are sufficiently informed. Although hospital nurses and doctors sense the difference between a patient who had visited the center before and who had not the stimulus for subsequent survey could be how the patients perceive the differentness of info provided within the center and out of it. The survey results could be a stimulus for establishing new centers not only within Vysočina but also in Austria. In the relation with the dissertation results and on the basis of info provided in the center when the patients assessed the info as sufficient for them an informing prospectus was created. The dissertation could serve as an informational material for students not only in the nursing profession.

This diploma thesis called "Need of non-pharmacology pain relieving for patients afflicted by Arthritis rheumatic" has been created as a next step of a previous pilot research. Respondents of the previous investigation expressed their interest in non-pharmacology pain relieving. It is very important to relief pain caused by Arthritis rheumatic as chronic disease in balance manner by usage of both approaches pharmacology and non-pharmacology. The goal of this study has been to investigate possibilities of non-pharmacology pain relieving among patients afflicted by Arthritis rheumatic and how these patients cooperate with nurses. The research comprises following goals: 1. Ascertain, whether patients afflicted by Arthritis rheumatic prefer pharmacology to non-pharmacology pain relieving. 2 Ascertain, which method of non-pharmacology pain relieving is mostly used. 3 Ascertain the interest of patients to relief pain by usage of non-pharmacology methods. 4 Ascertain the nurses' influence on the non-pharmacology pain relieving. The theoretical part is focused on pathophysiology of the disease, the issue of chronic pain and quality of patient life. The second, empirical part, is dedicated not only to actions of the investigation itself, but also its characteristics and results. There were issued 130 questionnairs for qualitative analisis. 102 of 130 questionairs has been retrieved and filled in properlly. Data from questionairs was compiled and hypotheses were tested by usage of Chi-squared test. 3 hypotheses have been built: H1 Patients afflicted by Arthritis rheumatic prefer pharmacology pain relieving to non-pharmacology. This hypothesis H1 was affirmed only for patients in acute stage of Arthritis rheumatic. H2 Patients afflicted by Arthritis rheumatic prefer thermotherapy to rehabilitation when considering non-pharmacology pain relieving. This hypothesis H2 was not confirmed due to low number of respondents. H3 Patients achieved higher education degree express higher interest in non-farmacology pain relieving. Higher education degree of patients has only impact on method in acute stage of Arthritis rheumatic. Qualitative investigation was carried out by half-structured interview with 7 respondents working on inpatient department and also outpatient department. Several statements could be concluded out of qualitative investigation: Nurses have been persistently educating by passive or active participation on various seminars or congresses. They have been trying to convey information about non-pharmacology treatment to their patients. They cobnsider non-pharmacology treatment as a indiscerptible part of a treatment. Investigation results will ne provided to all collaborative departments to support improvement in cooperation between nurses and patients.

Rheumatoid Arthritis (RA) is a chronic auto-immune disease that causes in ammation in the joints. Left uncontrolled, this prolonged in ammation can lead to pain and structural damage, resulting in erosions to the bones and total breakdown of the surrounding cartilage. Structural joint damage, measured by plain radiographs, is an important outcome measure of RA. It provides an objective marker of disease activity to assess any improvements or failures of treatments in controlling for the disease. Increased long-term joint damage has been linked with increased functional disability and decreased quality of life for RA patients. While a range of studies have looked at radiographic outcomes from observational data, they tend to be restricted to historical cohorts, with little long-term data on how radiographic progression may have changed in line with changes in clinical management. Additionally, these studies have not used the appropriate statistical methods to account for non-normal data distributions and within-patient variation over time. As a result, the main aim of this thesis is to investigate the long-term progression of structural joint damage in patients with early RA. The speci c objectives were to; (1) investigate the current evidence base to identify common methods in measuring and analysing radiographic outcomes, (2) assess what statistical methods are most appropriate in modelling long-term radiographic data, (3) use these models to understand the natural progression of radiographic damage using data from two UK inception cohorts, and nally, (4) expand these models to investigate the long-term relationship of radiographic damage with two important clinical outcomes; disease activity and functional disability. The analysis is based on longitudinal data from two UK prospective, multi-centre, early RA observational cohorts. These cohorts represent two distinct eras in the management and treatment of RA, making them invaluable for investigating how key RA outcomes have progressed in clinical practice over time. Using multi-level count models, precise rates of radiographic progression for both cohorts are presented. The models look at how seropositive RA and increased disease activity are related to increased radiographic progression, and what impact this has on functional disability. The results show that rates of radiological damage have declined dramatically in recent years. Possible attributable factors to these declines include both milder disease and more e ective treatment strategies. Analysis of the earlier cohort (1986-2001) shows how seropositive RA and increased disease activity lead to clinically meaningful increases in radiological damage. Conversely, their impact on patients in the more recent cohort (2002-2011) suggest that their e ect on radiographic progression is reduced, where increases in radiological damage were not larger than clinically meaningful thresholds. This has large implications on the debate around the use of biologic therapies in patients with less severe RA. However more data is sorely needed, particularly long-term radiographic data from those patients on biologics treatments, before any de nitive conclusions can be made. The possible impact of these declines on functional disability appears to be relatively small. The analysis shows that radiographic damage is more strongly associated with functional disability in later disease, but there is little evidence to indicate that declines in radiographic damage has lead to large improvements in long-term functional disability. These ndings are explored within the framework of a dual-pathway model, which suggests that functional disability is caused by two distinct mechanisms, either structural joint damage, or through increased pain. Research so far has predominantly focused on pharmacological treatments in reducing in ammation. More research is needed to explore the role of psychosocial factors and pain perception in order to create a more holistic treatment programme for RA patients.

Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / O gênero Pterodon pertence à família das Fabaceae e inclui quatro espécies nativas do Brasil: P. emarginatus Vog., P. apparicioi Pedersoli, P. abruptus Benth. e a espécie objeto deste estudo P. polygalaeflorus Benth.. Seus frutos são utilizados pela medicina popular devido às propriedades antirreumática, analgésica, anti-inflamatória, dentre outros. O objetivo deste trabalho foi avaliar a espécie Pterodon polygalaeflorus quanto ao seu potencial anti-inflamatório, antiartrítico e toxicológico, através da análise de seus efeitos em modelos in vitro e in vivo. Os extratos EEPpg, EHPpg e EDPpg reduziram (p<0,01) a produção in vitro de NO, por macrófagos ativados por LPS, com baixa citotoxicidade e diminuíram a celularidade (p<0,05) no exsudato inflamatório no modelo de inflamação in vivo conhecido como air pouch. O extrato mais ativo (EHPpg) foi selecionado e submetido a fracionamento em coluna de sílica gel 60 gerando quatro frações. Todas as frações (Fr I-Fr IV) reduziram: a produção de NO (p<0,001) por macrófagos ativados, com baixa ou nenhuma citotoxicidade; a migração de macrófagos in vitro (p<0,01, ensaio de wound healing) e in vivo (p<0,05, peritonite induzida por tioglicolato); e a proliferação de esplenócitos estimulados com Con A (p<0,001). As frações III e IV, mais ativas nos ensaios anteriores, mostraram ação antiartrítica (com 0,02 mg/kg), utilizando o modelo in vivo de artrite induzida por adjuvante completo de Freund (AIA), demonstrada por redução: do índice de edema de pata em 23,7% (Fr III) e 43,95% (Fr IV); das lesões histopatológicas na região tíbio-tarsal típicas de articulações com AIA (p< 0,05); do peso e celularidade do linfonodo e do baço, mas não da celularidade da medula óssea; das subpopulações de linfócitos (CD4+, CD8+ e CD19+) nos linfonodos inguinais, porém com significativo aumento das subpopulações ativadas CD4+CD69+, redução da CD8+CD69+ e aumento de CD19+CD69+ (apenas na Fr III); e redução da população de macrófagos no baço (p<0,001). As frações III e IV mostraram ação imunossupressora em nível celular e molecular, reduzindo (p<0,001) os níveis do mRNA da iNOS, assim como as citocinas IL-1&#946;, TNF-&#945; e IL-10, em nível de mRNA e proteína, em cultura de macrófagos. A expressão do receptor CD14, em macrófagos estimulados por LPS (31,74%), foi inibida apenas pela Fr III. A osteoclastogênese foi inibida pelas frações III e IV, sugerindo uma ação antiartrítica das frações em nível de diferenciação celular para osteoclastos (inibição). Este efeito pode resultar da inibição da expressão do fator de transcrição NFATc1, no caso da Fr III. Por fim, as frações Fr III e Fr IV não apresentaram toxicidade subaguda, potencial mutagênico (teste de Ames) ou genotóxico (teste do micronúcleo). A ausência de toxicidade in vivo das frações ficou demonstrada pela ausência de alteração no peso corporal e de órgãos, nas concentrações séricas de creatinina, ácido úrico, triglicerídeos, colesterol, ALT e ALP, ou de CYP1A1 e GSTs no fígado. Análises fitoquímicas (GC-MS e TLC) mostraram uma grande variedade de terpenos nas frações III e IV, sendo majoritários os furano-diterpenos derivados do vouacapano. O diterpeno isolado, Ppg-01, presente nas frações III (5,12%) e IV (18,47%), reduziu a produção in vitro de NO e o edema de pata induzido por carragenina (p<0,001). Em conjunto, os dados sugerem que o Ppg-01 esteja contribuindo para as ações anti-inflamatórias e imunomoduladoras das frações III e IV, e que estas propriedades estejam associadas aos efeitos antiartríticos observados.

Orientadora: Profa. Dra. Daniele Ribeiro de Araújo / Dissertação (mestrado) - Universidade Federal do ABC, Programa de Pós-Graduação em Biossistemas, 2016. / O naproxeno (NPX) é um anti-inflamatório não-esteroidal utilizado no tratamento da artrite reumatóide, osteoartrites e artroses, tanto como terapia única ou em associações farmacoterapêuticas. No entanto, sua administração por via oral está relacionada a efeitos adversos importantes (como toxicidade para o trato gastrintestinal). Nesse contexto, o desenvolvimento de novas formulações para liberação modificada de NPX possibilitaria a redução do número de injeções, aumentando a adesão ao tratamento. A incorporação de nanopartículas constituídas por quitosana-tripolifosfato (QT-TPP) em hidrogéis a base de poloxamers (PL) com diferentes valores de balanço hidrofílicolipofílico, favorece a formação de um sistema duplo de liberação com propriedades termoreversíveis, temperatura de gelificação (transição sol-gel) e razão de liberação do fármaco diferenciadas. Este trabalho visa o desenvolvimento (preparação e caracterização), a avaliação dos perfis de liberação in vitro e da citotoxicidade de novas formulações para liberação controlada de NPX incorporado em nanopartículas QT-TPP dispersas em hidrogéis termorreversíveis, visando a administração por via intra-articular, para o tratamento da artrite. Os diferentes hidrogéis foram preparados a base de poloxamer 407, (18 e 20 %), isolados ou associados ao poloxamer 403 (2 %). Nanopartículas constituídas de QT e TPP foram preparadas, caracterizadas e, posteriormente, dispersas nos hidrogéis termorreversíveis. Para isso, foram realizados estudos da interação fármaco-micelas e fármaconanopartículas (diâmetro hidrodinâmico, índices de polidispersão, eficiência de encapsulação e potencial zeta), para verificar a compatibilidade entre os componentes das formulações, bem como inferir sobre a estabilidade coloidal das mesmas; ii) a determinação da temperatura e da variação de entalpia relacionadas à micelização dos copolímeros isolados e de seus sistemas binários, antes e após a dispersão das nanopartículas, por calorimetria diferencial exploratória; iii) determinação da temperatura de transição sol-gel; iv) avaliação dos perfis de liberação in vitro do fármaco aplicandose diferentes modelos matemáticos. A dispersão das nanopartículas CS/TPP nos hidrogéis alterou as transições de fases e a termoreversibilidade das formulações, especialmente para aquelas com menor concentração de polímero. Por outro lado, a adição de nanopartículas contendo NPX, aos hidrogéis, aumentou as temperaturas relativas à micelização e a variação de entalpia. As análises reológicas mostraram perfis viscoelásticos e os resultados dos ensaios de liberação in vitro de NPX sugerem a contribuição de mecanismos como difusão e relaxamento das cadeias poliméricas. Por fim, com os ensaios de viabilidade celular observou-se a baixa citotoxicidade das formulações, apontando-as como sistemas híbridos promissores para liberação modificada de naproxeno. / Naproxen (NPX) is a non-steroidal anti-inflammatory used in the treatment of rheumatoid arthritis, osteoarthritis and arthrosis, either as sole therapy or pharmacotherapeutic associations. However, the oral administration is related to significant adverse effects (how toxicity to the gastrointestinal tract). Thus, in order to reduce the systemic side effects, intraarticular (IA) injection has been proposed as alternative. In this context, the development of new formulations for modified release NPX brings the possibility to reduce the number of injections, increasing the treatment adherence. The incorporation of nanoparticles of QT-TPP in poloxamers (PL) with different hydrophilic-lipophilic balance values, favors the formation of a dual release system of thermosensitive properties, gelation temperature (Tsol-gel) and due to the different drug release. This work aims the development, evaluation of the release profile in vitro and cytotoxicity of new formulations for controlled release of NPX-QT-TPP nanoparticles dispersed in thermosensitives hydrogels, aiming the injection intraarticular for treatment of arthritis. The different hydrogels were prepared based on poloxamer 407 (18 and 20%), isolated or associated with the poloxamer 403 (2%). Nanoparticles made of QT and TPP were prepared, characterized, and dispersed in the thermosensitives hydrogels. Therefore, studies of drug-micelle interaction and drug-nanoparticles were performed (hydrodynamic diameter, polydispersity, encapsulation efficiency and zeta potential), to verify the compatibility between the components of the formulations, as well as inferences about the colloidal stability of such; ii) determining the temperature and enthalpy change related to micellization of the isolated copolymers and their binary systems before and after dispersion of nanoparticles, by differential scanning calorimetry; iii) determination of the sol-gel transition temperature; iv) evaluation of the release profiles in vitro of the drug by applying different mathematical models. The dispersion of NanoQT/TPP nanoparticles in hydrogels changed phase transitions and thermosensitive formulations, especially those with lower concentrations of polymer. Moreover, the incorporation of NanoQT/TPP containing NPX to the hydrogels increased the micellization temperatures and enthalpy variation. Rheological analyzes showed viscoelastic rprofile and NPX in vitro release assays showed the contribution of mechanisms such as diffusion and relaxation of the polymer chains. Finally, cell viability assays showed low toxicity, indicating the hydrogels as promising hybrid systems for NPX modified release.