Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-05-18T11:28:47Z No. of bitstreams: 1 mirelleneryhenrique.pdf: 3757005 bytes, checksum: f81cbf74fa9d4f4e52254d53c930d9aa (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-07-01T18:23:14Z (GMT) No. of bitstreams: 1 mirelleneryhenrique.pdf: 3757005 bytes, checksum: f81cbf74fa9d4f4e52254d53c930d9aa (MD5) / Made available in DSpace on 2016-07-01T18:23:14Z (GMT). No. of bitstreams: 1 mirelleneryhenrique.pdf: 3757005 bytes, checksum: f81cbf74fa9d4f4e52254d53c930d9aa (MD5) Previous issue date: 2012-03-29 / Este estudo investigou a presença de cândida em manifestações bucais de pacientes com doença renal crônica em pré-diálise, em hemodiálise e em indivíduos saudáveis, utilizando-se de parâmetros clínicos e microbiológicos. Foram selecionados 34 pacientes portadores de doença renal crônica que foram divididos em dois grupos: GE1 – 20 em pré-diálise e; GE2 – 14 em hemodiálise. Para o terceiro grupo (GE3), foram selecionados aleatoriamente 14 indivíduos saudáveis que estavam aguardando atendimento odontológico em clínica particular. Todos os participantes selecionados passaram por uma anamnese e os mesmos responderam a alguns questionamentos sobre a saúde de um modo geral e sobre hábitos particulares de alimentação e hábitos nocivos. A verificação das manifestações bucais foi realizada de forma visual e devidamente fotografada e anotada. Durante o exame intrabucal, nas áreas que apresentavam diagnóstico sugestivo de candidíase, foi coletado material com swab para exame microbiológico, utilizando-se a técnica da Alça Calibrada, onde o meio foi o CHROMagarTM Cândida, para detectar o crescimento de três tipos de cândida: C. albicans, C. krusei e C. tropicalis. Para obtenção dos resultados foi utilizada a prova de Qui-Quadrado, ANOVA (com Post-Hoc LSD) e o teste t de Student, todos com nível de significância de p < 0,05. Das manifestações bucais diagnosticadas, aquela mais frequentemente encontrada foi a “Língua fissurada” com 78,6% no GE2; 60,0% no GE1 e 26,7% no GE3. A “Língua saburrosa” foi a segunda manifestação bucal mais observada, sendo 71,4% no GE2 e 45,0% no GE1. O GE2 foi o grupo que mais apresentou manifestações bucais. Com relação ao diagnóstico visual de candidíase nas manifestações bucais, o GE2 foi prevalente com 28,6%, seguido do GE1 com 25,0%. A C. albicans esteve presente com 17,6% nos pacientes com DRC, sendo 10,0% para o GE1 e 28,6% para o GE2. No GE3 não foi observada a presença de C. albicans. Desta forma, pode-se concluir que nas manifestações bucais observadas, o microrganismo C. albicans esteve presente nos pacientes com doença renal crônica em pré-diálise e hemodiálise, porém sem significância para afirmar que o mesmo seja oportunista para desencadear a candidíase. / This study investigated the presence of oral manifestations of candidiasis in patients with chronic kidney disease in predialysis, hemodialysis and healthy subjects, using the clinical and microbiological parameters. We selected 34 patients with chronic kidney disease who were divided into two groups: EG1 – 20 in pre-dialysis; EG2 – 14 on hemodialysis. For the third group (EG3) were randomly selected 14 healthy subjects who were waiting for dental care in private practice. All selected participants underwent an interview and they answered some questions about health in general and on particular habits of diet and harmful habits. Verification of oral manifestations was performed visually and duly photographed and recorded. During the examination intraoral areas with the diagnosis suggestive of candidiasis, swab material was collected for microbiological examination, using the technique of calibrated loop where the medium was CHROMagarTM Candida, to detect the growth of three types of bleach: C. albicans, C. krusei and C. tropicalis. To obtain the results we used the chi-square test, ANOVA (with post-hoc LSD) and Student's t test, all with a significance level of p < 0.05. Of oral lesions diagnosed, that was the most frequently found "Fissured tongue" with 78.6% in EG2, 60.0% and 26.7% in the EG1 and EG3. The "Tongue furred" was the second most frequent oral manifestation, being 71.4% and 45.0% in the EG2 and EG1. The EG2 was the group that presented oral manifestations. With respect to visual diagnosis of candidiasis in the oral manifestations, the EG2 was prevalent in 28.6%, followed by EG1 with 25.0%. The C. albicans was present in 17.6% in patients with chronic kidney disease, and to GE1 10.0% and 28.6% for EG2. EG3 was not observed in the presence of C. albicans. Thus, it can be concluded that oral manifestations observed in the microorganism C. albicans was present in patients with chronic kidney disease in predialysis and hemodialysis, but no significance to claim that it is opportunistic to trigger candidiasis.

Magister Scientiae Dentium - MSc(Dent) / Diabetes is often associated with a number of medical complications as a result of the metabolic changes taking place systemically. There is considerable evidence it is associated with oral complications including among others, gingivitis, periodontal disease, xerostomia, oral candidiasis, dental caries, periapical abscesses, lichen planus,burning mouth syndrome and an altered taste sensation (Lamster et al. 2008; Skamagas et al. 2008; Vernillo, 2003). The aim of the present study was to compare the oral health status in diabetic and non-diabetic patients with regards to their oral health problems, oral pathology and self perceived quality of life.A comparative cross-sectional study to determine the common oral complications prevalent in diabetics and non-diabetics was carried out in KwaZulu-Natal, at Prince Mshiyeni Memorial, EThekwini District, Umlazi. The study sample consisted of 150 diabetic patients and 150 non-diabetic patients attending the hospital. The oral health status was assessed clinically for each patient and recorded prior to the interview. The DMFT, plaque index and appearance of marginal gingiva were used to assess oral health status. An intra-oral examination was carried out to identify oral pathology lesions and other oral problems. Patients were then interviewed on the self perceived quality of life and the impact that diabetes has had on their lives. Plaque Index and DMFT were significantly higher among the diabetic group as opposed to the non-diabetic group. Periodontal disease was observed in more than half of the diabetic group as opposed to only 14.7% of the non-diabetic group. Except for tooth decay, the diabetic patients had more oral health problems compared to the non-diabetic group. More than half of the diabetic group reported having xerostomia compared to only 7.3% of the non-diabetic group. Altered taste sensation was also more prevalent in the diabetic group. In general, the diabetic group demonstrated a higher prevalence of oral pathology lesions and medical diabetes complications compared to the non-diabetic group.The self perceived quality of life was said to have deteriorated in 92% of diabetic subjects due to concomitant diabetic complications and 75% of this group indicated that they were not satisfied with their current quality of life.Diabetic patients were significantly less likely to perceive their quality of life as very satisfied after having adjusted for age and gender variables (OR=0.0188; CI: 0.0059-0.0594). Furthermore, diabetic patients were almost 6 times more likely to perceive themselves as “not satisfied” with their quality of life (QOL) as compared to non-diabetic patients.Individuals with diabetes exhibited poorer oral health when compared to non-diabetics.They exhibited a higher DMFT and had a significantly higher average number of missing teeth compared to the non-diabetic group. Special care needs to be given to diabetic patients because of the associated complications to improve their quality of life. A more detailed and in-depth study that utilises a diabetes-specific quality of life instrument may provide a more accurate way of determining the quality of life as well as periodontal disease in patients.

Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED, APS-I) is exceptionally common in Finland. Most patients have chronic oral candidiasis since childhood. Thus, most patients receive repeated courses of antifungals throughout their life. Eleven of our patients (31.4%) have become colonized with Candida albicans with decreased sensitivity to fluconazole. A total of 43 isolates of C. albicans from 23 APECED patients isolated during the years 1994 to 2004 were divided into 2 groups: fluconazole-susceptible dose-dependent (MIC, 16-32 μg/mL, 18 isolates) and fluconazole-susceptible (MIC ≤8 μg/mL, 25 isolates) groups. Antifungal activity of amphotericin B, echinocandins, and azoles was determined by the Clinical and Laboratory Standards Institute M27-A2 methodology. All isolates were highly susceptible to amphotericin B and echinocandins. Posaconazole and voriconazole were active against all isolates. Our data suggest that topical amphotericin B could continue to be a safe and active drug for daily administration for APECED patients. Posaconazole, voriconazole, and echinocandins may be useful in some complicated cases.

Ziel der vorliegenden Arbeit war anhand eines Tiermodells zu untersuchen, 1. ob eine Dosis-Wirkungsbeziehung zwischen der Keimmenge und der Entstehung einer oralen C. albicans Infektion besteht, 2. welche zelluläre Immunantwort auf definierte inokulierte Keimmengen stattfindet, 3. ob sich die Adhärenz von C. albicans an murine Epithelzellen durch Spaltprodukte von Muzin (Glykopeptide) verhindern läßt. Material und Methode: Immunkompetente Inzuchtmäuse (Balb/c) (n=27) und Mäuse mit kombiniertem B- und T-Zelldefekt (SCID) (n=30) wurden mit Keimmengen von10^4 bis 10^8 C. albicans-Zellen/10 mikrol des Stammes DSM 3454 oral inokuliert. Darüber hinaus wurden Balb/c Mäuse (n=8) mit 10^8 C. albicans-Zellen in Kombination mit Glykopeptiden und SCID Mäuse (n=8) mit 10^5 C. albicans-Zellen mit Glykopeptiden inokuliert. Eine Zungenhälfte wurde histologisch mittels Periodic-Acid-Schiff (PAS) Reaktion auf Invasion von Hyphen untersucht. Die andere Hälfte wurde mittels Immunperoxidase-Technik auf die Verteilung immunkompetenter Zellen (CD4, CD13, ICAM-1, E-Selectin, CD74, CD80, CD86, CD103) im Epithel und subepithelialen Bindegewebe untersucht. Ergebnisse: Eine Woche post inoculationem fanden sich weder bei Balb/c noch bei SCID Mäusen klinische Zeichen einer oralen Candidiasis. Die histologischen Ergebnisse mittels PAS-Methode zeigten jedoch, daß eine Inokulationsmenge von 10^8 C. albicans-Zellen bei Balb/c Mäusen und 10^8 Keime bei SCID Mäusen zu einer Infektion der Zungenmukosa führte. Das Ausmaß der immunologischen Reaktion war abhängig von der Inokulationsdosis sowie vom Immunstatus der Tiere. Die Ergebnisse der Inokulation von 10^8 C. albicans-Zellen zusammen mit Glykopeptiden zeigten bei 2/8 Balb/c Mäusen eine Hypheninvasion in das Zungenepithel. Bei 0/8 SCID Mäusen wurde nach Inokulation von 10^4 C. albicans-Zellen zusammen mit Glykopeptiden eine Hypheninvasion in das Zungenepithel beobachtet. Die immunhistochemischen Ergebnisse zeigten, daß die Reaktionen des Wirtes auf die Gabe der Keim-Glykopeptidlösung denen ohne Inokulation entsprachen. Schlußfolgerung: Obwohl bei den eingesetzten C. albicans-Mengen keine klinisch manifeste orale Candidiasis vorhanden war, fanden sich in beiden Tierstämmen inapparente Infektionen des Zungenepithel (Hypheninvasion), die immunologische Reaktionen der Zungenmukosa auslösten. Da nach Inokulation von C. albicans-Zellen zusammen mit Glykopeptiden weniger häufig Infektionen nachgewiesen werden konnten als bei Inokulation derselben Keimmenge ohne Glykopeptide und Nebenwirkungen dieser antiadhäsiven Wirkstoffe bisher nicht nachgewiesen wurden, wäre der unterstützende Einsatz von Muzinen oder deren Spaltprodukten bei Patienten mit erhöhtem Candidiasisrisiko zu erwägen. / This study applied an animal model to address the following questions: 1. Does a dose/effect relationship exist between C. albicans load and the emergence of an oral C. albicans infection, 2. which cellular immune response takes place following inoculation with defined pathogen loads, 3. is it possible to inhibit C. albicans adhesion to murine epithelium cells through the local application of mucine metabolites (glycopeptides). Material and methods: Immunocompetent inbred mice (Balb/c) (n=27) and mice with combined B- and T-cell defects (SCID) (n=30) were orally inoculated with pathogen loads between 10^4 and 10^8 C. albicans cells/10 microl (strain DSM 3454). Moreover, Balb/c mice (n=8) were inoculated with 10^8 C. albicans cells in combination with glycopeptides; SCID mice (n=8) were inoculated with 10^5 cells, also in combination with glycopeptides. One half of the tongue tissue was histochemically examined with the Periodic Acid Schiff (PAS) Method for displaying the invasion of hyphae. The other half of the tissue was examined by immune peroxidase technique for analysing the distribution of immunocompetent cells (CD4, CD13, ICAM-1, E-Selectin, CD74, CD80, CD86, CD103) in the epithelial layers and subepithelial connective tissue. Results: One week following the inoculation, neither group's tissue showed clinical signs of oral candidiasis. Following histochemical preparation (PAS-Reaction) the tongue mucosa showed signs of infection (hyphae) with the inoculation dose of 10^8 C. albicans cells in the case of Balb/c mice and a load of 10^5 pathogens in the case of SCID mice. The extent of the immunologic reaction depended both on the inoculation dose given to the animals and on their immune status. The results of an inoculation of 10^8 C. albicans cells in combination with glycopeptides showed hyphae invasion of the tongue epithelium in 2/8 Balb/c mice. Following an inoculation of 10^5 pathogens in combination with glycopeptides hyphae invasion could be demonstrated in 0/8 SCID mice. The results of immunohistochemical studies showed that the host's reaction to combined glycopeptide-pathogen-inoculation correspond to the reaction without inoculation. Conclusion: Despite the lack of clinical signs of oral candidiasis in neither group's tissue, non-apparent infections of the tongue epithelium were evident leading to immunologic reactions of the tongue mucosa. Inoculation of C. albicans cells in combination with glycopeptides resulted in decreased infection rate compared to a corresponding inoculation dose without glycopeptides. As no side effects have been documented for the oral application of these antiadhesive agents, their use as complimentary therapy for patients at an increased risk for oral candidiasis should be considered.

A nistatina (NYS) é o fármaco de primeira escolha no tratamento da candidíase oral, que frequentemente acomete mais os indivíduos imunocomprometidos e pacientes com outras desordens (diabetes não tratada, neoplasias, imunodeficiências). No mercado brasileiro, a NYS é encontrada na forma de suspensão oral aquosa, onde o procedimento para sua administração consiste em bochechar o medicamento. Apesar de haver a indicação de que se mantenha o contato direto entre fármaco e a mucosa oral, na qual se encontra a Candida spp., o que aumentaria expressivamente o sucesso terapêutico, a suspensão não apresenta tal propriedade. Assim, a NYS que é fármaco com ação efetiva contra a candidíase oral, é considerada pertencente à Classe IV do Sistema de Classificação Biofarmacêutica, ou seja, apresenta baixa solubilidade e baixa permeabilidade. A baixa solubilidade pode comprometer sua disponibilidade na cavidade oral, e consequentemente, sua ação farmacológica. Diante desse quadro, o objetivo do presente trabalho foi o desenvolvimento de dispersões sólidas de NYS para o tratamento da candidíase oral, e sua posterior incorporação em gel mucoadesivo oral, favorecendo a formulação no local de ação. As dispersões sólidas são sistemas farmacêuticos, onde um fármaco pouco solúvel em água encontra-se dispersado em um carreador, no estado sólido. Os carreadores normalmente são hidrofílicos, o que permite que esses sistemas sejam empregados para aumentar a solubilidade aquosa do fármaco. Assim, foram desenvolvidas as dispersões sólidas de NYS, pelo método de eliminação do solvente, empregando como carreadores, lactose, HPMC, poloxamer 407 e poloxamer 188. Essas foram submetidas à caracterização por análise térmica, usando os ensaios de calorimetria exploratória diferencial (DSC) e termogravimetria/termogravimetria derivada (TG/DTG). Dentre essas dispersões sólidas, aquelas que se mostraram com comportamento térmico sugerindo a formação de um novo \"sistema\", foram analisadas por meio de ensaio de solubilidade. Dessa forma, a formulação NYS DS G2 (49) se destacou, pois apresentou maior solubilidade em água (4,484 mg/mL); em pH 5,5 (4,249 mg/mL) e em pH 7,0 (4,293 mg/mL), ou seja, houve um aumento de 1,426 vezes em água; 4,227 vezes em pH 5,5; e 2,743 vezes em pH 7,0. Essa formulação foi, por fim avaliada por difração de raio-X e espectroscopia de infravermelho com transformada de Fourier, técnicas que corroboraram com a análise térmica quanto à indicação de formação da dispersão sólida. Por sua vez, essa dispersão sólida foi incorporada em 4 bases de géis mucoadesivos de carbopol ® 934 PNF, alterando apenas a concentração do polímero (0,5; 1,0; 1,5; 2,0 %p/p). Foi observado que a liberação de NYS DS G2 (49) foi superior, quando comparada à liberação de NYS MP a partir do gel, e através do ensaio de mucoadesão, percebeu-se que os géis desenvolvidos apresentaram propriedades mucoadesivas compatíveis com relatos na literatura, independentemente da quantidade de carbopol ® empregada. As características reológicas foram distintas, e foi observado que as formulações Gel A e Gel B, que possuem menor quantidade de polímero, tiverem um indicativo de comportamento de fluido newtoniano, diferente dos demais, o que pode não ser desejado para esse tipo de forma farmacêutica tópica e semi-sólida. Ao final desse trabalho, pode-se concluir que foi possível desenvolver um sistema farmacêutico na forma de dispersão sólida com maior solubilidade que a NYS pura, e sua incorporação em uma forma farmacêutica mucoadesiva, e que a liberação da NYS na forma DS foi muito superior que o fármaco na forma \"convencional\", o que permite que a NYS esteja mais disponível na cavidade oral, e também junto à mucosa bucal, o que levaria a efeito farmacológico mais efetivo do antifúngico. / Nystatin (NYS) is the drug of first choice in the treatment of oral candidiasis that most often affect immunocompromised individuals, and patients with other disorders. In the Brazilian market, NYS is found in the form of aqueous oral suspension, a medication used in the form of mouthwash. Although there is an indication to maintain direct contact between the drug and the oral mucosa, where Candida spp. is found, as well as where therapeutic success would significantly be increased, the suspension has no such property. Thus, the NYS is an effective drug against oral candidiasis, and belongs to Class IV of the Biopharmaceutical Classification System, it has low solubility and low permeability. The low solubility can compromise its availability in the oral cavity, and consequently, its pharmacological action. Given this situation, the objective of this work was the development of solid dispersions of NYS for the treatment of oral candidiasis, and its subsequent incorporation into oral mucoadhesive gel, in order to facilitate its action. Solid dispersions are pharmaceutical systems, in which a solid drug poorly soluble in water is dispersed in a carrier. These carriers are usually hydrophilic, and this allows the systems to be employed in order to increase the aqueous solubility of the drug. Thus, the solid NYS dispersions were developed by the solvent evaporation method, employing lactose, HPMC, poloxamer 407 and poloxamer 188 as carrier. These samples were subjected to characterization by thermal analysis, using differential scanning calorimetry (DSC) and thermogravimetry / derivative thermogravimetry (TG / DTG). Among these solid dispersions, those samples which showed a specific thermal behavior suggesting the formation of new \"system\" were analyzed by solubility test. Thus, the NYS DS G2 formulation (49) stood out, once it showed greater solubility in water (4.484 mg/mL); at pH 5.5 (4.249 mg/mL) and pH 7.0 (4.293 mg/mL), in other words, an increase of 1,426 times in water; 4,227 times at pH 5.5; and 2,743 times at pH 7.0. This formulation was finally evaluated by X-ray diffraction, infrared spectroscopy with Fourier transform, techniques that corroborate the thermal analysis, indicating the formation of the solid dispersion. On the other hand, this solid dispersion was incorporated into 4 Carbopol ® 934 PNF mucoadhesive gels, with a variation of the polymer concentration. It was observed that NYS is improved of delivery from the gels, employing mucoadhesion test, and was also observed that the gels have mucoadhesive properties consistent with reports in the literature. However, the rheological characteristics are different, and it was observed that the Gel A and Gel B formulations, which has a lower amount of polymer behaved as a Newtonian fluid, which may not be desired for this type of topical gel. As conclusion, it was possible to develop a pharmaceutical system in the form of solid dispersion with greater solubility than the pure NYS, and their incorporation in a mucoadhesive dosage form and the release of NYS as DS was far superior wherein the drug in the \"conventional\" manner, which allows the NYS is longer available in the oral cavity, and also adjacent to the buccal mucosa, leading to more effective pharmacological effect of the antifungal agent.

A nistatina (NYS) é o fármaco de primeira escolha no tratamento da candidíase oral, que frequentemente acomete mais os indivíduos imunocomprometidos e pacientes com outras desordens (diabetes não tratada, neoplasias, imunodeficiências). No mercado brasileiro, a NYS é encontrada na forma de suspensão oral aquosa, onde o procedimento para sua administração consiste em bochechar o medicamento. Apesar de haver a indicação de que se mantenha o contato direto entre fármaco e a mucosa oral, na qual se encontra a Candida spp., o que aumentaria expressivamente o sucesso terapêutico, a suspensão não apresenta tal propriedade. Assim, a NYS que é fármaco com ação efetiva contra a candidíase oral, é considerada pertencente à Classe IV do Sistema de Classificação Biofarmacêutica, ou seja, apresenta baixa solubilidade e baixa permeabilidade. A baixa solubilidade pode comprometer sua disponibilidade na cavidade oral, e consequentemente, sua ação farmacológica. Diante desse quadro, o objetivo do presente trabalho foi o desenvolvimento de dispersões sólidas de NYS para o tratamento da candidíase oral, e sua posterior incorporação em gel mucoadesivo oral, favorecendo a formulação no local de ação. As dispersões sólidas são sistemas farmacêuticos, onde um fármaco pouco solúvel em água encontra-se dispersado em um carreador, no estado sólido. Os carreadores normalmente são hidrofílicos, o que permite que esses sistemas sejam empregados para aumentar a solubilidade aquosa do fármaco. Assim, foram desenvolvidas as dispersões sólidas de NYS, pelo método de eliminação do solvente, empregando como carreadores, lactose, HPMC, poloxamer 407 e poloxamer 188. Essas foram submetidas à caracterização por análise térmica, usando os ensaios de calorimetria exploratória diferencial (DSC) e termogravimetria/termogravimetria derivada (TG/DTG). Dentre essas dispersões sólidas, aquelas que se mostraram com comportamento térmico sugerindo a formação de um novo \"sistema\", foram analisadas por meio de ensaio de solubilidade. Dessa forma, a formulação NYS DS G2 (49) se destacou, pois apresentou maior solubilidade em água (4,484 mg/mL); em pH 5,5 (4,249 mg/mL) e em pH 7,0 (4,293 mg/mL), ou seja, houve um aumento de 1,426 vezes em água; 4,227 vezes em pH 5,5; e 2,743 vezes em pH 7,0. Essa formulação foi, por fim avaliada por difração de raio-X e espectroscopia de infravermelho com transformada de Fourier, técnicas que corroboraram com a análise térmica quanto à indicação de formação da dispersão sólida. Por sua vez, essa dispersão sólida foi incorporada em 4 bases de géis mucoadesivos de carbopol ® 934 PNF, alterando apenas a concentração do polímero (0,5; 1,0; 1,5; 2,0 %p/p). Foi observado que a liberação de NYS DS G2 (49) foi superior, quando comparada à liberação de NYS MP a partir do gel, e através do ensaio de mucoadesão, percebeu-se que os géis desenvolvidos apresentaram propriedades mucoadesivas compatíveis com relatos na literatura, independentemente da quantidade de carbopol ® empregada. As características reológicas foram distintas, e foi observado que as formulações Gel A e Gel B, que possuem menor quantidade de polímero, tiverem um indicativo de comportamento de fluido newtoniano, diferente dos demais, o que pode não ser desejado para esse tipo de forma farmacêutica tópica e semi-sólida. Ao final desse trabalho, pode-se concluir que foi possível desenvolver um sistema farmacêutico na forma de dispersão sólida com maior solubilidade que a NYS pura, e sua incorporação em uma forma farmacêutica mucoadesiva, e que a liberação da NYS na forma DS foi muito superior que o fármaco na forma \"convencional\", o que permite que a NYS esteja mais disponível na cavidade oral, e também junto à mucosa bucal, o que levaria a efeito farmacológico mais efetivo do antifúngico. / Nystatin (NYS) is the drug of first choice in the treatment of oral candidiasis that most often affect immunocompromised individuals, and patients with other disorders. In the Brazilian market, NYS is found in the form of aqueous oral suspension, a medication used in the form of mouthwash. Although there is an indication to maintain direct contact between the drug and the oral mucosa, where Candida spp. is found, as well as where therapeutic success would significantly be increased, the suspension has no such property. Thus, the NYS is an effective drug against oral candidiasis, and belongs to Class IV of the Biopharmaceutical Classification System, it has low solubility and low permeability. The low solubility can compromise its availability in the oral cavity, and consequently, its pharmacological action. Given this situation, the objective of this work was the development of solid dispersions of NYS for the treatment of oral candidiasis, and its subsequent incorporation into oral mucoadhesive gel, in order to facilitate its action. Solid dispersions are pharmaceutical systems, in which a solid drug poorly soluble in water is dispersed in a carrier. These carriers are usually hydrophilic, and this allows the systems to be employed in order to increase the aqueous solubility of the drug. Thus, the solid NYS dispersions were developed by the solvent evaporation method, employing lactose, HPMC, poloxamer 407 and poloxamer 188 as carrier. These samples were subjected to characterization by thermal analysis, using differential scanning calorimetry (DSC) and thermogravimetry / derivative thermogravimetry (TG / DTG). Among these solid dispersions, those samples which showed a specific thermal behavior suggesting the formation of new \"system\" were analyzed by solubility test. Thus, the NYS DS G2 formulation (49) stood out, once it showed greater solubility in water (4.484 mg/mL); at pH 5.5 (4.249 mg/mL) and pH 7.0 (4.293 mg/mL), in other words, an increase of 1,426 times in water; 4,227 times at pH 5.5; and 2,743 times at pH 7.0. This formulation was finally evaluated by X-ray diffraction, infrared spectroscopy with Fourier transform, techniques that corroborate the thermal analysis, indicating the formation of the solid dispersion. On the other hand, this solid dispersion was incorporated into 4 Carbopol ® 934 PNF mucoadhesive gels, with a variation of the polymer concentration. It was observed that NYS is improved of delivery from the gels, employing mucoadhesion test, and was also observed that the gels have mucoadhesive properties consistent with reports in the literature. However, the rheological characteristics are different, and it was observed that the Gel A and Gel B formulations, which has a lower amount of polymer behaved as a Newtonian fluid, which may not be desired for this type of topical gel. As conclusion, it was possible to develop a pharmaceutical system in the form of solid dispersion with greater solubility than the pure NYS, and their incorporation in a mucoadhesive dosage form and the release of NYS as DS was far superior wherein the drug in the \"conventional\" manner, which allows the NYS is longer available in the oral cavity, and also adjacent to the buccal mucosa, leading to more effective pharmacological effect of the antifungal agent.

Orientador: Ana Claudia Pavarina / Banca: Marco Antonio Compagnoni / Banca: Ana Lucia Machado / Banca: Juliana Campos Junqueira / Banca: Karin Hermana Neppelenbroek / Resumo: Estes estudos in vivo tiveram como objetivos: 1 - avaliar a efetividade da terapia fotodinâmica (PDT) na inativação de Candida albicans em um modelo murino de candidose bucal; 2 - comparar a eficácia clínica e microbiológica da PDT com a da terapia antifúngica tópica (nistatina suspensão oral) no tratamento da estomatite protética, assim como identificar e determinar a prevalência das espécies de Candida dessa patologia. Na primeira investigação, camundongos foram imunossuprimidos com injeções subcutâneas de prednisolona. Tetraciclina foi fornecida na água de beber para promover alteração da microbiota bucal dos camundongos. Os animais foram sedados com injeção de cloridrato de clorpromazina e um swab oral embebido em uma suspensão de C. albicans (107 UFC/mL) foi esfregado na cavidade bucal dos animais. Quatro dias após a inoculação, a PDT foi realizada no dorso lingual utilizando administração tópica de Photogem® a 400, 500 ou 1000 mg/L e, após 30 minutos, iluminação com 305 J/cm² de luz de LED a 455 ou 630 nm. Posteriormente, a quantidade de fungos viáveis foi determinada (UFC/mL) e analisada pelos testes de ANOVA e Holm- Sidak (P < 0,05). Os camundongos foram sacrificados, e as línguas foram removidas e processadas para avaliação histológica de presença de fungos e reação inflamatória. No estudo clínico, pacientes (n = 40) foram aleatoriamente atribuídos a um dos seguintes grupos de 20 indivíduos cada; grupo NYS: pacientes receberam tratamento tópico com nistatina (100.000 UI) quatro vezes ao dia por 15 dias e; grupo PDT: prótese total superior e o palato dos pacientes foram borrifados pelo Photogem® a 500 mg/L e, após 30 minutos de incubação, iluminado por luz de LED a 455 nm (37,5 e 122 J/cm2, respectivamente) três vezes por semana durante 15 dias. Culturas micológicas de amostras das próteses e das mucosas palatinas e fotografias... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: These in vivo studies evaluated 1 - the effectiveness of photodynamic therapy (PDT) on the inactivation of Candida albicans in a murine model of oral candidosis; 2 - compared the clinical and mycological efficacy of PDT with that of topical antifungal therapy (nystatin oral suspension) for the treatment of denture stomatitis (DS) as well as to identify and determine the prevalence of Candida species in DS. In the first investigation, mice were immunosupressed with subcutaneous injections of prednisolone. Tetracycline hydrochloride on drinking water was managed to change the oral microflora. They were kept in a sedative state after injection of chlorpromazine chloride and a suspension of C. albicans (107 CFU/mL) was swabbed in the oral cavity. Four days after oral infection, PDT was performed on the dorsum of the tongue using a topical administration of Photogem® at 400, 500 or 1000 mg/L and, after 30 minutes, illumination with 305 J/cm² of LED light at 455 or 630 nm. Then, the number of surviving yeast was determined (CFU/mL) and analyzed by ANOVA and Holm- Sidak tests (P < 0.05). Animals were sacrificed, the tongues surgically removed and processed for histological evaluation of yeast presence and inflammatory reaction. In the clinical trial, patients (n = 40) were randomly assigned to one of two groups of 20 subjects each; NYS group: patients received topical treatment with nystatin (100,000 IU) four times daily for 15 days; PDT group: maxillary denture and palate of patients were sprayed with 500 mg/L of Photogem® and, after 30 min of incubation, illuminated by LED light at 455 nm (37.5 and 122 J/cm2, respectively) three times a week for 15 days. Mycological cultures taken from dentures and palates and standard photographs of the palates were performed at baseline (day 0), at the end of the treatment (day 15) and at the follow-up (days 30, 60 and 90 after the beginning... (Complete abstract click electronic access below) / Doutor

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-03-06T11:14:23Z No. of bitstreams: 1 arquivo total.pdf: 786129 bytes, checksum: 1e08eca0cb38b5c895cff1f2cbb20f5c (MD5) / Made available in DSpace on 2017-03-06T11:14:24Z (GMT). No. of bitstreams: 1 arquivo total.pdf: 786129 bytes, checksum: 1e08eca0cb38b5c895cff1f2cbb20f5c (MD5) Previous issue date: 2016-12-13 / Oral candidiasis is a fungal infection caused by yeasts of the genus Candida, with C. albicans being the main pathogen. The emergence of resistant species against the available therapeutic arsenal as well as its adverse effects has driven new research on natural products. In view of this problem, the present study sought to investigate the antifungal activity of essential oil (OE) obtained from the leaves of Melissa officinalis L (lemon balm), isolated and associated with antifungal agents licensed on C. albicans strains. Two strains of the American Type Culture Collection (ATCC) and eight from the oral cavity (LM) were selected. The microdilution and sowing of subcultures were used to determine, respectively, the Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (CFM). The antifungigram was performed using the disc diffusion technique with the antifungal agents nystatin, amphotericin B, miconazole and fluconazole, evaluating them in isolation and associated with OE. MIC and MFC ranged from 64 to 128 μg/mL and 256 to 512 μg/mL, respectively. The presence of resistance of the strains to the antifungal agents nistatin (LM-38 and LM-65), amphotericin B (ATCC-76485, LM-3A, LM-38, LM-62 and LM -65) and fluconazole (LM-1A, LM- 1B, LM-38, LM-62 and LM-65). In the association tests, the results showed the influence of the essential oil on the antifungal activity, with synergism (S), indifference (I) and antagonism (A) being observed. The percentages found for each antifungal in the order (S, I and A) were: nystatin (30%, 30% , 40%), amphotericin B (30%, 50%, 20%), fluconazole (20% ,40% , 40%) and miconazole (100%, 0,0). In view of the above, it is concluded that the OO of M. Officinalis may become a promising product in the fight against oral candidiasis, since it has a strong antifungal activity against Candida albicans and that the use of this product associated with the licensed antifungal can, in some situations, potentiate its effectiveness of use in treatment. / A candidíase oral é uma infecção fungica causada por leveduras do gênero Candida, sendo a espécie C. albicans apontada como principal patógeno. O surgimento de espécies resistentes frente ao arsenal terapêutico disponível assim como seus efeitos adversos tem impulsionado novas pesquisas voltadas para os produtos naturais. Diante dessa problemática o presente trabalho buscou investigar a atividade antifúngica do óleo essencial (OE) obtido das folhas de Melissa officinalis L (erva-cidreira), isolado e associado a antifúngicos licenciados sobre cepas de C. albicans. Para tanto, foram selecionadas duas cepas da American Type Culture Collection (ATCC) e oito provenientes de cavidade oral (LM). Utilizou-se a técnica da microdiluição e semeadura de subcultivos para determinar, respectivamente, a Concentração Inibitória Mínima (CIM) e a Concentração Fungicida Mínima (CFM). O antifungigrama foi realizado a partir da técnica de difusão de disco com os antifúngicos nistatina, anfotericina B, miconazol e fluconazol avaliando-os de forma isolada e associada ao OE. A CIM e CFM variaram, respectivamente, de 64 a 128 μg/mL e 256 a 512 μg/mL. Quanto ao teste de sensibilidade de forma isolada observou-se a presença de resistência das cepas aos antifúngicos nistatina (LM-38 e LM-65), anfotericina B (ATCC- 76485, LM-3A, LM-38, LM-62 e LM-65) e fluconazol (LM-1A, LM-1B, LM-38, LM- 62 e LM-65). Nos ensaios de associação os resultados mostraram influência do óleo essencial sobre a atividade dos antifúngicos sendo observado sinergismo (S), indiferença (I) e antagonismo (A). As porcentagens encontradas para cada antifúngico, na ordem (S, I e A), foram: nistatina (30%, 30% e 40%), anfotericina B (30%, 50%, 20%), fluconazol (20%, 40% e 40%) e miconazol (100%,0,0). Perante o exposto conclui-se que o OE de M. Officinalis pode se tornar um produto promissor no combate a candidose oral, visto que apresenta forte atividade antifungica contra Candida albicans e que o uso desse produto associado à antifungicos licenciados pode, em algumas situações, potencializar sua efetividade de uso no tratamento.

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-03-06T13:18:25Z No. of bitstreams: 1 arquivo total.pdf: 1622677 bytes, checksum: 4910b306e46e741a75cc3d2047affa2c (MD5) / Made available in DSpace on 2017-03-06T13:18:25Z (GMT). No. of bitstreams: 1 arquivo total.pdf: 1622677 bytes, checksum: 4910b306e46e741a75cc3d2047affa2c (MD5) Previous issue date: 2015-08-17 / Introduction: Faced with limitations to the use of sodium hypochlorite in the disinfection of dental prostheses and the need to control fungal proliferation in these sites, it is necessary to study new substances for this purpose. Objectives: To evaluate the antifungal activity, mechanism of action, cytotoxicity and antibiofilm action of chloramine T (CAT) on Candida spp. The minimum inhibitory concentration (MIC) of the substance on Candida albicans, Candida tropicalis, Candida krusei and Candida glabrata was determined by the microdilution technique and the minimum fungicidal concentration (CFM) was calculated through the subculture in Sabouraud Dextrose Agar (ASD) . The growth inhibition kinetics of C. albicans were evaluated by the method of counting colony forming units (CFU) at different times and concentrations. A microculture of C. albicans was performed on fowl agar plus tween 80 to evaluate the possible alteration of micromorphology against different concentrations of the substance. The possible mechanism of action on wall and fungal cell membrane was verified by the determination of MIC in the presence of sorbitol and ergosterol respectively. The inhibition of the initial adherence of fungal cells, formation and reduction of C. albicans biofilm were evaluated after short (1 min) and prolonged (8 h) contact with the substance and formation of the biofilm was measured by absorbance at 600 nm, Transformed into scores referring to the percentage of inhibition obtained based on the values ​​of the control group. The cytotoxicity of the substance was evaluated by the hemolysis method. Nystatin and sodium hypochlorite were used as positive controls. A descriptive and inferential analysis was performed considering α = 5%. Results: The CIM75% found for CAT was 781.3 μg / mL and the CFM / MIC ratio suggests a fungicidal activity against most of the strains tested, with probable action on cell wall and membrane. The substance showed immediate and prolonged action on the kinetic test and caused reduction of the filamentous form and inhibition of chlamydoconidia. In the biofilm assay, it presented similar results to sodium hypochlorite for inhibition of initial adherence and formation of mature biofilm (p> 0.05) and was more effective in reducing mature biofilm in the short contact groups at MIC concentration x2 (24 H) and CIM x 4 (48 h) (p <0.05). Conclusion: CAT shows antifungal activity on Candida spp. And shows fungicidal action on most of the strains tested. Its action is immediate and prolonged in the inhibition of C. albicans growth and probably occurs both in the wall and in the cell membrane. CAT causes alterations in the micromorphology of C. albicans and has antibiofilm activity, being effective in inhibiting the initial adherence of fungal cells, as well as in the formation and reduction of biofilm. / Introducao: Diante das limitacoes para o uso do hipoclorito de sodio na desinfeccao de proteses dentarias e da necessidade do controle da proliferacao fungica nestes sitios, faz-se necessario o estudo de novas substancias para este fim. Objetivos: Avaliar a atividade antifungica, mecanismo de acao, citotoxicidade e acao antibiofilme da cloramina T (CAT) sobre Candida spp. Materiais e Metodos: Foi determinada a concentracao inibitoria minima (CIM) da substancia sobre Candida albicans, Candida tropicalis, Candida krusei e Candida glabrata pela tecnica da microdiluicao e calculada a concentracao fungicida minima (CFM) atraves do subcultivo em Agar Sabouraud Dextrose (ASD). Foi avaliada a cinetica de inibicao do crescimento de C. albicans pelo metodo de contagem de unidades formadoras de colonias (UFC) em diferentes tempos e concentracoes. Foi realizado microcultivo de C. albicans em agar fuba acrescido de tween 80 para avaliacao da possivel alteracao da micromorfologia frente a diferentes concentracoes da substancia. O possivel mecanismo de acao sobre parede e membrana celular fungica foi verificado atraves da determinacao da CIM na presenca, respectivamente, de sorbitol e ergosterol. A inibicao da aderencia inicial de celulas fungicas, formacao e reducao do biofilme de C. albicans foram avaliados apos contato curto (1 min) e prolongado (8 h) com a substancia e a formacao do biofilme foi mensurada atraves de absorbancia a 600 nm, transformada em escores referentes a porcentagem de inibicao obtida com base nos valores do grupo controle. A citotoxicidade da substancia foi avaliada pelo metodo da hemolise. Nistatina e hipoclorito de sodio foram utilizados como controles positivos. Foi realizada analise estatistica descritiva e inferencial, considerando α=5%. Resultados: A CIM75% encontrada para a CAT foi de 781,3 µg/mL e a relacao CFM/CIM sugere uma atividade fungicida frente a maioria das cepas testadas, com provavel acao em parede e membrana celulares. A substancia mostrou acao imediata e prolongada no teste de cinetica e provocou reducao da forma filamentosa e inibicao de clamidoconidios. No ensaio do biofilme, apresentou resultados semelhantes ao hipoclorito de sodio para inibicao da aderencia inicial e formacao do biofilme maduro (p>0,05) e foi mais efetiva na reducao do biofilme maduro nos grupos de contato curto na concentracao CIM x 2 (24 h) e CIM x 4 (48 h) (p < 0,05). Conclusao: A CAT apresenta atividade antifungica sobre Candida spp. e apresenta acao fungicida sobre a maioria das cepas testadas. Sua acao e imediata e prolongada na inibicao do crescimento de C. albicans e provavelmente ocorre tanto em parede quanto em membrana celular. A CAT causa alteracoes na micromorfologia de C. albicans e possui atividade antibiofilme, sendo efetiva na inibicao da aderencia inicial das celulas fungicas, bem como na formacao e reducao do biofilme.

Made available in DSpace on 2014-12-17T14:16:28Z (GMT). No. of bitstreams: 1 MarianaGD_DISSERT.pdf: 1350118 bytes, checksum: 5e25fd552ff8be29e71b6c0ae2fa383c (MD5) Previous issue date: 2011-02-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Despite Candida species are often human commensals isolated from various oral sites such as: tongue, cheek and palatal mucosa plus subgingival region, there are some properties linked to the organism commonly known as virulence factors which confer them the ability to produce disease. Oral candidiasis is one of the main oral manifestations reported in literature related to kidney transplant patients. The objectives of the present study were to identify and investigate virulence factors of yeasts isolated from the oral cavity of kidney transplant recipients admitted at the Hospital Universit?rio Onofre Lopes, in Natal RN. Seventy Candida species isolated from 111 kidney transplant recipients were investigated in this study. Identification of the isolates was performed by using the evidence of germ tube formation, hypertonic broth, tolerance to grow at 42?C, micromorphology and biochemical profiles. We observed a high rate of isolation of yeasts from the oral cavity of kidney transplant recipients (63.1%) being C. albicans was the most prevalent species. Oral candidiasis was diagnosed in 14.4% of transplant recipients. We evaluated virulence properties of the isolates regarding to: biofilm formation on polystyrene microplates as well as XTT reduction, adherence to acrylic resin and human buccal epithelial cells and proteinase activity. Most isolates were able to form biofilm by the method of adhesion to polystyrene. All isolates of Candida spp. remained viable during biofilm formation when analyzed by the method of XTT reduction. The number of CFU attached to the acrylic resin suggested high adherence for C. parapsilosis. C. albicans isolates showed higher median adherence to human buccal epithelial cells than non-C. albicans Candida isolates. Nevertheless, this difference was not statistically significant. C. dubliniensis showed low ability to adhere to plastic and epithelial cells and biofilm formation. Proteolytic activity was observed for all the isolates investigated, including the unique isolate of C. dubliniensis. There was a statistically significant association between proteinase production and the presence of oral candidiasis. Studies related to oral candidiasis in renal transplant recipients are limited to clinical and epidemiological data, but investigations concerning Candida spp. virulence factor for this group of individuals are still scarce. We emphasize the importance of studies related to virulence factors of yeasts isolated from this population to contribute to the knowledge of microbiological aspects of oral candidiasis / Apesar das leveduras do g?nero Candida serem frequentemente comensais humanos, isoladas de diferentes s?tios orais, incluindo l?ngua, mucosa jugal, mucosa palatal e regi?o subgengival, existem algumas propriedades ligadas a Candida spp., comumente denominadas fatores de virul?ncia, que lhes conferem a capacidade de produzir doen?a. Candid?ase bucal ? uma das principais manifesta??es orais citadas na literatura em rela??o aos pacientes transplantados renais. O objetivo deste estudo foi realizar identifica??o e avaliar os fatores de virul?ncia de leveduras isoladas da cavidade bucal de receptores de transplante renal que s?o acompanhados no Hospital Universit?rio Onofre Lopes, na cidade do Natal RN. Foram utilizadas 70 leveduras do g?nero Candida isoladas de 111 receptores de transplante renal. A identifica??o dos isolados foi realizada atrav?s das provas de forma??o de tubo germinativo, caldo hipert?nico, toler?ncia ? temperatura de 42?C, an?lise da micromorfologia e perfil bioqu?mico das leveduras. Observamos elevado ?ndice de isolamento de leveduras na cavidade bucal dos receptores de transplante renal (63,1%), havendo predom?nio de C. albicans. Candid?ase bucal foi diagnosticada em 14,4% dos transplantados. Avaliou-se o potencial de virul?ncia das leveduras atrav?s da forma??o de biofilme pelo m?todo de ader?ncia a microplaca de poliestireno, redu??o do XTT, habilidade de ader?ncia a corpos de prova de resina acr?lica e a c?lulas epiteliais bucais, bem como atividade de proteinase. A maioria dos isolados foi capaz de produzir biofilme pelo m?todo de ader?ncia ao poliestireno, determinada atrav?s de leitura em espectrofot?metro. Todos os isolados de Candida spp. permaneceram vi?veis durante a forma??o do biofilme pelo m?todo da redu??o do XTT. A contagem do n?mero de UFC aderidas ao corpo de prova demonstrou alta capacidade de ader?ncia de C. parapsilosis. Os isolados de C. albicans apresentaram maior mediana de ades?o ? c?lula epitelial bucal humana do que os isolados de C. n?o-C. albicans, contudo essa diferen?a n?o foi estatisticamente significativa. C. dubliniensis apresentou baixa capacidade de ader?ncia ao pl?stico e c?lulas epiteliais e forma??o do biofilme. Observamos atividade proteol?tica em todos os isolados pesquisados, inclusive o isolado de C. dubliniensis, e associa??o estatisticamente significativa entre a produ??o de proteinase e a presen?a de candid?ase bucal. Estudos relacionados ? candid?ase bucal em receptores de transplante renal limitam-se ? investiga??o de aspectos cl?nicos e epidemiol?gicos, n?o havendo dados concernentes a fatores de virul?ncia. Ressaltamos a import?ncia da realiza??o de estudos relacionados aos fatores de virul?ncia de leveduras isoladas nessa popula??o, a fim de que se aprofunde o conhecimento dos aspectos microbiol?gicos da candid?ase bucal / 2020-01-01