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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.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede.biblioteca.ufpb.br:tede/8858 |
| Date | 17 August 2015 |
| Creators | Ferreira, Gabriela Lacet Silva |
| Contributors | Castro, Ricardo Dias de |
| Publisher | Universidade Federal da Paraíba, Programa de Pós-Graduação em Odontologia, UFPB, Brasil, Odontologia |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFPB, instname:Universidade Federal da Paraíba, instacron:UFPB |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | -7411869720500764667, 600, 600, 600, 1726686701291727373, -2070498469879244349 |
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