Atividade antifúngica de nanofibras de policaprolactona/nistatina: estudo in vitro e in vivo / Antifungal activity of polycaprolactone/nystatin nanofibers: in vitro and in vivo study

Este estudo avaliou a eficácia in vitro e in vivo de mantas de nanofibras (NF) de policaprolactona (PCL) incorporadas com nistatina (NIS) no tratamento da estomatite protética (EP) em modelos animais. NF foram sintetizadas com diferentes concentrações de NIS, totalizando quatro soluções: PCL puro, PCL/NIS 0,045 g, PCL/NIS 0,090 g e PCL/NIS 0,225 g. A liberação da NIS foi analisada por espectroscopia Ultravioleta-Visível. A capacidade das mantas de inibirem o biofilme de Candida albicans, principal fator etiológico da EP, dividindo-se cinco grupos (N=5) compostos por um grupo com controle de células de C. albicans e com PCL puro, além das três concentrações de NIS. A seguir, foi analisada a viabilidade celular em queratinócitos humanos (HaCat) por meio do teste colorimétrico de resazurina. Cinco grupos foram divididos (N=10): controle celular, PCL puro e as três concentrações de NIS. Em modelos animais de ratos Wistar albinos (N=18), dispositivos palatinos (DP) de resina acrílica foram confeccionados simulando próteses totais e utilizados para a indução da EP. Para isso, DP contaminados com C. albicans foram cimentados na região molar da cavidade bucal dos animais e permaneceram em boca por 48 h. Após esse período, os DP foram removidos e os animais foram divididos em três grupos: (C) controle; (B1) com tratamento por mantas de PCL/NIS 0,045 g e (B2) PCL/NIS 0,225 g, com N=6. Então novos DP, livres de contaminação, foram cimentados na cavidade oral dos animais e permaneceu por mais 48 h. Após esse período, os animais foram eutanasiados, a contagem de UFC/ mL foi realizada e os palatos foram coletados para a análise histológica. A curva padrão de NIS obtida apresentou R2 de 0,99. As três concentrações de NF apresentaram liberação de NIS, com pico no tempo de 6 h e valores de 66,26 µg/ mL para PCL/NIS 0,045 g, de 333,87 µg/ mL para PCL/NIS 0,090 g e 436,51 µg/ mL para PCL/NIS 0,225 g, constantes até o fim do experimento. Os grupos com NIS reduziram em 2,5 log10 de crescimento do biofilme fúngico em relação aos grupos sem tratamento, Controle e PCL, sem diferença estatística significativa. Não foi observada citotoxicidade nas células HaCat, com viabilidade celular de 93,7% para PCL/NIS 0,045 g, 72,6% para PCL/NIS 0,090 g e 72,4% para PCL/NIS 0,225 g. A indução da EP nos três grupos foi possível e, porém, sem redução significativa na contagem de UFC/ mL de C. albicans nos grupos B1 e B2. Na análise histológica do grupo C pôde-se observar infiltração de hifas de Candida na camada queratinizada, presença de células inflamatórias formando micro abscessos e um discreto infiltrado inflamatório no tecido conjuntivo subjacente ao epitélio infectado. Nos grupos B1 e B2 não foram encontradas alterações epiteliais, concluindo-se que as NF demonstraram atividade antifúngica in vitro e foram efetivas na prevenção da penetração de hifas no tecido palatino de animais com DP (AU)

This study evaluated the in vitro and in vivo efficacy of nanofiber (NF) mats of polycaprolactone (PCL) incorporated with nystatin (NIS) in the treatment of denture stomatitis (DS) in animal models. NFs were synthesized with different concentrations of NIS, totaling four solutions: pure PCL, PCL/NIS 0.045 g, PCL/NIS 0.090 g, and PCL/NIS 0.225 g. The release of NIS was analyzed by Ultraviolet-Visible spectroscopy. The ability of the mats to inhibit Candida albicans biofilm, the main etiological factor of DS, was assessed by dividing five groups (N=5) composed of a group with C. albicans cell control and with pure PCL, in addition to the three concentrations of NIS. Next, cell viability in human keratinocytes (HaCat) was analyzed using the resazurin colorimetric test. Five groups were divided (N=10): cell control, pure PCL, and the three concentrations of NIS. In albino Wistar rat animal models (N=18), palatal devices (PD) made of acrylic resin were fabricated to simulate total prostheses and used to induce DS. For this, PD contaminated with C. albicans were cemented in the molar region of the animals' oral cavity and remained in the mouth for 48 hours. After this period, the PDs were removed, and the animals were divided into three groups: (C) control; (B1) treated with PCL/NIS 0.045 g mats, and (B2) PCL/NIS 0.225 g, with N=6. Then new, uncontaminated PDs were cemented in the animals' oral cavity and remained for another 48 hours. After this period, the animals were euthanized, UFC/ mL counts were performed, and the palates were collected for histological analysis. The standard NIS curve obtained showed an R2 of 0.99. The three concentrations of NF showed NIS release, with a peak at 6 h and values of 66.26 µg/ mL for PCL/NIS 0.045 g, 333.87 µg/ mL for PCL/NIS 0.090 g, and 436.51 µg/ mL for PCL/NIS 0.225 g, remaining constant until the end of the experiment. The groups with NIS reduced fungal biofilm growth by 2.5 log10 compared to the untreated groups, Control and PCL, with no significant statistical difference. No cytotoxicity was observed in HaCat cells, with cell viability of 93.7% for PCL/NIS 0.045 g, 72.6% for PCL/NIS 0.090 g, and 72.4% for PCL/NIS 0.225 g. Induction of DS in the three groups was possible; however, there was no significant reduction in UFC/ mL counts of C. albicans in groups B1 and B2. Histological analysis of group C revealed infiltration of Candida hyphae in the keratinized layer, presence of inflammatory cells forming micro abscesses, and a discreet inflammatory infiltrate in the connective tissue underlying the infected epithelium. No epithelial alterations were found in groups B1 and B2, concluding that NFs demonstrated in vitro antifungal activity and were effective in preventing hyphal penetration into palatal tissue in animals with PD.(AU)

Efecto de la terapia combinada en la vaginosis bacteriana recurrente en mujeres del Eje Cafetero, Colombia / Effect of combined therapy on recurrent bacterial vaginosis in women from the Coffee Region, Colombia, 2017-2020

Objetivo: Evaluar el efecto de la terapia combinada, en la vaginosis bacteriana recurrente, en mujeres del Eje Cafetero (Colombia). Métodos: Estudio observacional, en 189 mujeres mayores de 18 años, con diagnóstico de vaginosis bacteriana recurrente siguiendo el puntaje de Nugent. Se les suministró terapia combinada (inducción oral con 500 mg de metronidazol por siete días más 600 mg de ácido bórico vaginal una vez al día durante veintiún días, seguido de 500 mg de nifuratel y 200 000 UI de nistatina vaginal por seis días, cada mes, durante seis meses); quienes asistieron a consulta entre 2017 y 2020. Se hizo seguimiento clínico y de laboratorio utilizando el puntaje de Nugent, a la semana de finalizado el tratamiento oral, al mes, a los tres, seis, nueve y doce meses. Se utilizó estadística descriptiva. Resultados: La edad media de las participantes fue 34,17 ± 5,31 años. La tasa de curación después de la terapia inicial fue 80,42 % (a la semana), al mes: 82,01 %, a los tres meses: 91,53 %, a los seis: 90,47 %, a los nueves: 90,47 % y a los doce meses: 86,24 %. Al finalizar el estudio, la tasa de fracaso fue 9,52 %. No hubo eventos adversos graves; la tasa de satisfacción fue elevada (90,47 %). Conclusiones: La terapia combinada en la vaginosis bacteriana recurrente mejoró la cura clínica y microbiológica y disminuyó el riesgo de recurrencia a los doce meses de seguimiento. Se requiere validación de los presentes resultados en un ensayo clínico controlado aleatorizado(AU)

Objective: To evaluate the effect of combined therapy in recurrent bacterial vaginosis in women from the Coffee Region (Colombia). Methods: Observational study, in 189 women over 18 years, diagnosed with recurrent bacterial vaginosis following the Nugent score. They were given combination therapy (oral induction with 500 mg metronidazole for seven days plus 600 mg vaginal boric acid once daily for twenty-one days, followed by 500 mg nifuratel and 200,000 IU vaginal nystatin for six days, every month, for six months); who attended consultation between 2017 and 2020. Clinical and laboratory follow-up was performed using Nugent's score, one week after the end of oral treatment, at one month, three, six, nine and twelve months. Descriptive statistics were used. Results: The mean age of participants was 34.17 ± 5.31 years. The cure rate after initial therapy was 80.42% (at week), at month: 82.01 %, three months: 91.53 %, six: 90.47 %, nine: 90.47 % and at twelve months: 86.24 %. At the end of the study, the failure rate was 9.52%. There were no serious adverse events; the satisfaction rate was high (90.47 %). Conclusions: Combination therapy in recurrent bacterial vaginosis improved clinical and microbiological cure and decreased the risk of recurrence at twelve months follow-up. Validation of the present results is required in a randomized controlled clinical trial(AU)

Histoplasmosis: informe de un caso / Histoplasmosis: a case report

Objetivo: Describir el diagnóstico y el tratamiento interdisciplinario de un caso clínico de histoplasmosis. Caso clínico: Un paciente masculino de 39 años, con antecedentes de consumo de drogas, alcohol, tabaquismo crónico y VIH+ sin adherencia al tratamiento, acudió al Servicio de Odontología por una interconsulta del Servicio de Clínica Médica para la evaluación de lesiones erosivas en paladar duro y blando, reborde alveolar anterior, dorso lingual y lesión tumoral en encía anterosuperior. El diagnóstico definitivo se obtuvo por medio del análisis de muestras de biopsia transbronquial, lavado broncoalveolar y biopsia de lesión en piel. El paciente recibió tratamiento sistémico con antimicóticos (anfotericina B e itraconazol según esquema) y tratamiento local con colutorio de clorhexidina al 0,12% y 100.000 UI de nistatina en suspensión. Al momento del alta, presentaba una considerable mejoría de su estado general y de las lesiones orales, con disminución de sintomatología dolorosa. Se indicó turno para control a los 7 días de forma ambulatoria, al cual el paciente no asistió. El abordaje interdisciplinario y el análisis de los diferentes aspectos socioeconómicos, culturales, ambientales y sistémicos del paciente facilitaron el diagnóstico temprano de la enfermedad (AU)

Aim: To describe the diagnosis and interdisciplinary treatment of a clinical case of histoplasmosis. Clinical case: 39-year-old male patient with a history of drug abuse, alcohol, and chronic smoking, HIV+ without treatment compliance, attends the dental department referred by the medical department for the diagnosis of erosive lesions in the hard and soft palate, anterior alveolar ridge, lingual dorsum and tumor lesion in the anterosuperior gingiva. The definitive diagnosis was obtained by the analysis of transbronchial biopsy, bronchoalveolar lavage and skin lesion biopsy. The patient received systemic treatment with antifungals (amphotericin b, itraconazole according to protocol), and local treatment with 0.12% chlorhexidine mouthwash and 100,000 IU nystatin suspension. At the time of medical discharge, the patient presented a considerable improvement in his general condition and of the oral lesions with a reduced pain. A 7 days recall was prescribed, however the patient failed to attend. The interdisciplinary approach to the patient and the analysis of the different socio-economic, cultural, environmental and systemic aspects of the patient facilitates the early diagnosis of the disease (AU)

Efficacy and safety of prophylactic antifungal Agents in preventing invasive fungal infection and mortality among infants weighing less than 1500 grams: A meta-analysis

@#<p style="text-align: justify;"><strong>Background. </strong>Preterm infants with very low birth weight are at increased risk of invasive fungal infections. Preventive strategies are needed to improve their clinical course and survival.</p><p style="text-align: justify;"><strong>Objectives. </strong>To assess the efficacy and safety of antifungal agents as prophylaxis in controlling invasive fungal infection and mortality in very low birth weight (VLBW) and extremely low birth weight (ELBW) infants in neonatal intensive care units.</p><p style="text-align: justify;"><strong>Methods. </strong>We searched MEDLINE (PubMed), Cochrane databases, Google Scholar, Trip database, Herdin, and ClinicalTrials.gov without language restriction and publications from January 1988 to May 2021. We included randomized controlled trials or controlled clinical trials that compared the effect of prophylactic oral or systemic antifungal agents versus placebo in preterm infants < 37 weeks age of gestation and with birth weight lower than 1500 grams. We conducted a meta-analysis using RevMan 5.4.1 and certainty of evidence rating using GRADEpro software.</p><p style="text-align: justify;"><strong>Results. </strong>A total of 14 studies (including 3,001 preterm infants with VLBW) were included. We found that prophylactic use of nystatin significantly reduced the incidence of invasive fungal infections (IFI) (pooled RR 0.16; 95% CI 0.11, 0.23; 4 RCTs, N = 1295; P < 0.00001; moderate certainty evidence) in preterm infants compared to placebo but had no significant effect on the mortality (RR 0.87; 95% CI 0.62, 1.23; 4 RCTs, N = 1295; P = 0.43; low certainty evidence). Similarly, fluconazole decreased the incidence of IFI (RR 0.38; 95% CI 0.28, 0.53; P = 0.02) and showed statistically significant reduction in mortality (RR 0.78; 95% CI 0.61, 0.99; RCTs, N = 1484; P = 0.04; high certainty evidence). The comparison of the two antifungals showed a trend favoring fluconazole, however the difference was not statistically significant in decreasing IFI (RR 1.60; 95% CI 0.68, 3.77; P = 0.28) and mortality (RR 1.62; 95% CI 0.76, 3.45; P = 0.21).</p><p style="text-align: justify;"><strong>Conclusion.</strong> Administration of antifungal prophylaxis proves to be beneficial and can probably decrease invasive fungal infection and mortality. The evidence showed that Fluconazole is superior as antifungal prophylaxis compared to placebo while there is no significant difference between fluconazole and nystatin in decreasing fungal infection and mortality among preterm neonates.</p>

Beneficial effects of three natural products for the treatment of denture stomatitis: a randomized clinical trial / Efeitos benéficos de três produtos naturais para o tratamento da estomatite protética: um ensaio clínico randomizado

Aim: To evaluate the effect of three natural antifungal agents combined with routine denture care on the treatment of DS, using a quantitative mycological culture analysis. Methods: Thirty denture wearers with denture stomatitis DS were treated using five substances: sterile distilled water (G1), nystatin oral suspension (G2), 20% alcoholic extract propolis (G3), Punica granatumLinné gel (G4), and Uncaria tomentosa gel (G5). The substances were used 3 times a day for 14 days. Quantitative mycological culture analysis of samples collected from the palatal mucosa was performed at three stages: before treatment (T0), after 14 days of treatment (T1), and 30 days after treatment completion (T2). Data were evaluated using Kruskal-Wallis and Friedman tests (p < 0.05). Results: Palatal mucosa intragroup analysis showed a significant reduction of Candida CFU/mL values for all groups at T1 compared to T0 (p < 0.05). However, they did not present statistical differences when comparing T1 and T2 (p > 0.05). The intergroup analysis demonstrated that there are no statistical differences, regardless of the evaluation time (p > 0.05). Conclusion:The natural products tested showed a satisfactory result on DS treatment, which proved to be equivalent to conventional topical therapy with nystatin and to treatment using only regular oral hygiene procedures.

Objetivo: Avaliar o efeito de três antifúngicos naturais combinados com o cuidado rotineiro com próteses dentárias no tratamento da EP, por meio de uma análise quantitativa de cultura micológica. Métodos: Trinta usuários de próteses dentárias com EP foram tratados com cinco substâncias: água destilada estéril (G1), suspensão oral de nistatina (G2), extrato alcoólico de própolis 20% (G3), gel Punica granatum L. (G4) e gel Uncaria tomentosa (G5). As substâncias foram utilizadas 3 vezes ao dia durante 14 dias. A análise micológica quantitativa das amostras coletadas da mucosa palatina foi realizada em três etapas: antes do tratamento (T0), após 14 dias do tratamento (T1) e 30 dias após o término do tratamento (T2). Os dados foram avaliados pelos testes de Kruskal-Wallis e Friedman (p < 0,05). Resultados: A análise intragrupo da mucosa palatina mostrou uma redução significativa dos valores de Candida UFC/mL para todos os grupos em T1 em comparação com T0 (p < 0,05). No entanto, não apresentaram diferenças estatísticas na comparação de T1 e T2 (p > 0,05). A análise intergrupos demonstrou que não há diferenças estatísticas, independentemente do tempo de avaliação (p > 0,05). Conclusão: Os produtos naturais testados apresentaram resultado satisfatório no tratamento da EP, sendo equivalente à terapia tópica convencional com nistatina e ao tratamento apenas com procedimentos rotineiros de higiene bucal.

Surface morphology and in vitro leachability of soft liners modified by the incorporation of antifungals for denture stomatitis treatment

Abstract Objective: To evaluate the surface morphology and in vitro leachability of temporary soft linings modified by the incorporation of antifungals in minimum inhibitory concentrations (MIC) for Candida albicans biofilm. Methodology:Specimens of soft lining materials Softone and Trusoft were made without (control) or with the addition of nystatin (Ny), miconazole (Mc), ketoconazole (Ke), chlorhexidine diacetate (Chx), or itraconazole (It) at their MIC for C. albicans biofilm. The surface analyses were performed using Confocal laser scanning microscopy after 24 h, 7 days, or 14 days of immersion in distilled water at 37ºC. In vitro leachability of Chx or Ny from the modified materials was also measured using Ultraviolet visible spectroscopy for up to 14 days of immersion in distilled water at 37ºC. Data (µg/mL) were submitted to ANOVA 1-factor/Bonferroni (α=0.05). Results: Softone had a more irregular surface than Trusoft. Morphological changes were noted in both materials with increasing immersion time, particularly, in those containing drugs. Groups containing Chx and It presented extremely porous and irregular surfaces. Both materials had biexponential release kinetics. Softone leached a higher concentration of the antifungals than Trusoft (p=0.004), and chlorhexidine was released at a higher concentration than nystatin (p<0.001). Conclusions: The surface of the soft lining materials changed more significantly with the addition of Chx or It. Softone released a higher concentration of drugs than Trusoft did, guiding the future treatment of denture stomatitis.

Efecto antifúngico del aceite de canela en comparación con nistatina, como tratamiento de Candida Albicans Huánuco 2017 / Antifungal effect of cinnamon oil compared to nystatin, as a treatment for Candida Albicans Huánuco 2017

La presente investigación aborda el campo clínico del comportamiento de las cepas del hongo candida albicans, causantes de infecciones oportunistas en pacientes inmuno suprimidos, así como de las alternativas terapéuticas empleando medicina natural (Aceite de Canela) así como de fármacos (Nistatina), en el tratamiento anti fúngico. En el Capítulo I, se hace referencia al planteamiento del problema, señalando la formulación del mismo, los objetivos y la trascendencia de la investigación. El Capítulo II se hace referencia al marco teórico, precisando los conocimientos teóricos, epistemológicos, enfoques, teorías, modelos y fundamentos paradigmáticos correspondientes a la problemática evidenciada en los hallazgos de laboratorio respecto a las cepas y sus efectos en el ser humano. En el Capítulo III se señala el marco metodológico indicando el tipo de investigación, población, muestra; así mismo de las técnicas e instrumentos de recolección de datos, procesamiento y análisis de la información obtenida durante el desarrollo de la investigación. En el Capítulo IV se señalan los resultados o hallazgos de la investigación; finalmente en el Capítulo V se precisa la discusión y contrastación de los resultados. El estudio concluyó en que existen diferencias significativas entre el aceite de canela y la nistatina, empleadas en el tratamiento anti fungicida de la candida albicans, en concentraciones al 50% y 100%, pero no existen diferencias significativas diferencias significativas al 25% del aceite de canela.

Anadenanthera Colubrina vell Brenan: anti-Candida and antibiofilm activities, toxicity and therapeutical action

Abstract: We evaluated the antifungal and antibiofilm potential of the hydroalcoholic extract of bark from Anadenanthera colubrina (vell.) Brenan, known as Angico, against Candida spp. Antifungal activity was evaluated using the microdilution technique through the Minimum Inhibitory and Fungicide Concentrations (MIC and MFC). The antibiofilm potential was tested in mature biofilms formed by Candida species and analyzed through the counting of CFU/mL and scanning electron micrograph (SEM). In vivo toxicity and therapeutic action was evaluated in the Galleria mellonella model. The treatment with the extract, in low doses, was able to reduce the growth of planktonic cells of Candida species. MIC values range between 19.5 and 39 µg/mL and MFC values range between 79 and 625 µg/mL. In addition was able to reduce the number of CFU/mL in biofilms and to cause structural alteration and cellular destruction, observed via SEM. A. colubrina showed low toxicity in the in vivo assay, having not affected the viability of the larvae at doses below 100mg/kg and high potential in the treatment of C. albicans infection. Considering its high antifungal potential, its low toxicity and potential to treatment of infections in in vivo model, A. colubrina extract is a strong candidate for development of a new agent for the treatment of oral candidiasis.

Avaliação in vitro do potencial erosivo e cariogênico de antifúngicos tópicos / In vitro evaluation of erosive and cariogenic potential of topical antifungal agents

Introdução: a candidíase é uma infecção fúngica oportunista, causada pela proliferação e disseminação de espécies de Candida, que pode acometer a cavidade oral. Dentre os antifúngicos mais utilizados e de uso tópico, a nistatina é considerada o medicamento de primeira escolha. Objetivo: avaliar as propriedades físico-químicas de diferentes marcas de nistatina disponíveis no mercado, incluindo o pH, a acidez total titulável (ATT) e a determinação de sólidos solúveis totais (SST). Metodologia: trata-se de um estudo experimental in vitro, constituído por uma amostra de oito diferentes marcas de nistatina em suspensão oral de uso tópico. Foi analisado o potencial erosivo e cariogênico dessas soluções mediante a determinação de pH, ATT e SST (°Brix). Resultados: no tocante ao pH, verificou-se que a média obtida foi de 6,05 (± 0,66). Dois dos medicamentos analisados (marcas A e H) apresentaram pH abaixo do crítico para a dissolução do esmalte dental. Quanto à ATT das soluções, os valores variaram de 1,9 a 14,53 mL para atingir o pH neutro, indicando que as marcas B, C e E podem levar mais tempo para ser neutralizadas em razão da quantidade de solução necessária. A análise do °Brix revelou que a marca H apresentou o maior teor de açúcares em sua composição (44,9%). Conclusão: a formulação de nistatina da marca H apresentou pH endógeno mais crítico e percentual de sólidos solúveis totais elevado, sendo, portanto, a medicação com maior fator de risco para o desenvolvimento de cárie e erosão dentária, devendo ser consideradas as doses e frequências de uso, bem como os hábitos de higiene oral do paciente

Introduction: candidiasis is an opportunistic fungal infection caused by the proliferation and spread of Candida species that can affect the oral cavity. Among the most commonly used topical antifungal agents, nystatin is considered the first choice drug. Methodology: to evaluate the physical and chemical properties of different brands of nystatin available in the market, including pH, titratable acidity and determination of total soluble solids. Results: Regarding pH, it was verified that the mean obtained was 6.05 (± 0.66). Two of the analyzed drugs (A and H) presented pH below that considered critical for the dissolution of dental enamel. As for the titratable total acidity of the solutions, values ranged from 1.9 to 14.53 mL to reach neutral pH, indicating that the B, C and E marks may take longer to neutralize because of the amount of solution required. The analysis of ° Brix revealed that the H mark had the highest sugar content in its composition (44.9%). Conclusion: Nystatin brand H presented the worst indices in terms of endogenous pH and total sugar percentage, being therefore the medication with the highest risk factor for the development of caries and dental erosion.

Interactions between Terpinen-4-ol and Nystatin on biofilm of Candida albicans and Candida tropicalis

Abstract The aim of this study was to evaluate the antifungal activity of Terpinen-4-ol associated with nystatin, on single and mixed species biofilms formed by Candida albicans and Candida tropicalis, as well as the effect of terpinen-4-ol on adhesion in oral cells and the enzymatic activity. The minimum inhibitory concentrations and minimum fungicide concentrations of terpinen-4-ol and nystatin on Candida albicans and Candida tropicalis were determined using the microdilution broth method, along with their synergistic activity ("checkerboard" method). Single and mixed species biofilms were prepared using the static microtiter plate model and quantified by colony forming units (CFU/mL). The effect of Terpinen-4-ol in adhesion of Candida albicans and Candida tropicalis in coculture with oral keratinocytes (NOK Si) was evaluated, as well as the enzymatic activity by measuring the size of the precipitation zone, after the growth agar to phospholipase, protease and hemolysin. Terpinen-4-ol (4.53 mg mL-1) and nystatin (0.008 mg mL-1) were able to inhibit biofilms growth, and a synergistic antifungal effect was showed with the drug association, reducing the inhibitory concentration of nystatin up to 8 times in single biofilm of Candida albicans, and 2 times in mixed species biofilm. A small decrease in the adhesion of Candida tropicalis in NOK Si cells was showed after treatment with terpinen-4-ol, and nystatin had a greater effect for both species. For enzymatic activity, the drugs showed no action. The effect potentiated by the combination of terpinen-4-ol and nystatin and the reduction of adhesion provide evidence of its potential as an anti-fungal agent.

Resumo O objetivo desse estudo foi avaliar a atividade antifúngica do Terpinen4-ol associado à nistatina em biofilmes simples e misto, formados por Candida albicans e Candida tropicalis, bem como o efeito do terpinen-4-ol na adesão em células orais e atividade enzimática. As concentrações inibitórias mínimas e as concentrações fungicidas mínimas do terpinen-4-ol e da nistatina em Candida albicans e Candida tropicalis foram determinadas pelo método de microdiluição em caldo, juntamente com a atividade sinérgica (método do tabuleiro de "xadrez"). Biofilmes simples e misto foram preparados usando o modelo de placa de microtitulação estática e quantificados por unidades formadoras de colônias (CFU/mL). O efeito do Terpinen-4-ol na adesão de Candida albicans e Candida tropicalis em co-cultura com queratinócitos orais (NOK Si) foi avaliado, bem como a atividade enzimática, medindo o tamanho da zona de precipitação, após o crescimento em ágar fosfolipase, protease e hemolisina. O terpinen-4-ol (4.53 mg mL-1) e a nistatina (0,008 mg mL-1) conseguiram inibir o crescimento de biofilmes e um efeito antifúngico sinérgico foi demonstrado com a associação de fármaco, reduzindo a concentração inibidora de nistatina até 8 vezes em biofilme simpes de Candida albicans e 2 vezes em biofilme misto. Uma pequena diminuição na adesão de Candida tropicalis em células NOK Si foi mostrada após o tratamento com terpinen-4-ol e a nistatina teve um efeito maior para ambas as espécies. Para a atividade enzimática, as drogas não apresentaram ação. O efeito potencializado pela combinação de terpinen-4-ol e nistatina e a redução de adesão evidenciam seu potencial como agente anti-fúngico.

Prise en charge des vaginites dans les pays à ressources limitées d'Afrique subsaharienne. Quelle place pour l'association ternidazole, néomycine sulfate, nystatine et prednisolone ?

Contexte : La vaginite est un motif fréquent de consultation gynécologique. Dans les pays à ressources limitées, en l'absence de laboratoires d'analyses microbiologiques des sécrétions vaginales, le traitement est prescrit sur la base de l'évaluation clinique.Objectif général : Evaluer l'efficacité de notre pratique concernant la prise en charge des vaginites en situation de vie réelle, par la prescription de l'associationternidazole, néomycine sulfate, nystatine, prednisolone (Tergynan®).Méthodologie : Nous avons réalisé, dans trois structures sanitaires d'Abidjan, une étude prospective observationnelle sur cinq mois. Elle a inclus, de façon aléatoire, 233 patientes qui ont donné leur consentement écrit et signé et qui présentaient des signes cliniques évidents de vaginites. N'ont pas été incluses les patientes enceintes et allaitantes, celles qui présentaient des co-infections, des infections sexuellement transmissibles, le VIH, une hypersensibilité ou une idiosyncrasie à l'association ternidazole, néomycine sulfate, nystatine, prednisolone (Tergynan®). Les patientes ont été divisées en deux groupes : le groupe de référence (n = 200) pour lequel une analyse microbiologique des sécrétions vaginales était effectuée 2 à 4 jours avant la mise sous traitement et le groupe contrôle (n = 33) pour qui le traitement était débuté d'emblée. Les critères d'évaluation étaient la présence des symptômes cliniques et leur cinétique d'évolution, le bilan microbiologique réalisé 10 à 14 jours après le début du traitement et la survenue d'effets indésirables liés à l'utilisation du médicament (notamment les picotements ou les irritations locales). Les tests statistiques utilisés pour la comparaison des effectifs étaient le khi-deux et le Fisher exact avec un seuil de signification de 5% (p < 0,05).Résultats et discussion : Avant l'instauration du traitement, les leucorrhées pathologiques (178/233) et le prurit vulvaire (157/233) étaient les principaux symptômes rencontrés dans les deux groupes. La prédominance des leucorrhées pathologiques était très significativement différente dans le groupe référence versus le groupe contrôle (p = 0,001).Lors de la visite de contrôle, il n'y avait pas de différence significative entre les deux groupes concernant la présence d'une récidive (p = 0,99). Ainsi l'association ternidazole, néomycine sulfate, nystatine, prednisolone (Tergynan®) pourrait représenter une option thérapeutique intéressante chez les femmes présentant des signes cliniques évidents de vaginite, en l'absence d'analyse microbiologique préalable des sécrétions vaginales. L'association ternidazole, néomycine sulfate, nystatine, prednisolone (Tergynan®) a démontré son efficacité thérapeutique en cas de vaginite à Candida albicans (VC), de Vaginose Bactérienne (VB), de vaginite à Trichomonas vaginalis (VT), et même en cas d'infection mixte (VM). En effet il a été observé une amélioration significative et rapide (3 à 4 jours maximum) des symptômes (p = 0,001) et un faible taux de récidive biologiquement prouvée dans les deux groupes (3,4%). Durant toute la durée du traitement, les patientes n'ont déclaré aucune réaction allergique liée à l'utilisation de l'association ternidazole, néomycine sulfate, nystatine, prednisolone (Tergynan®) (picotements ou irritations locales) et 97,3% des patientes ont été globalement très satisfaites de l'utilisation de l'association ternidazole, néomycine sulfate, nystatine, prednisolone (Tergynan®).Conclusion : Dans nos pays à ressources limitées, sous-médicalisés, l'association ternidazole, néomycine sulfate, nystatine, prednisolone (Tergynan®) apparaît comme une option thérapeutique intéressante pour traiter les vaginites d'origine mycosique, parasitaire, bactériologique et mixte. Cette étude a montré que l'association ternidazole, néomycine sulfate, nystatine, prednisolone (Tergynan®) pouvait être administrée, en initiation de traitement des vaginites dès leur apparition sans besoin de procéder à des prélèvements préalables des sécrétions vaginales

Efecto antifúngico de diferentes concentraciones del extracto de Uncaria Tomentosa sobre Candida albicans: Estudio in vitro / Efeito antifúngico de diferentes concentrações hidroalcoólicas de UncariaTomentosa em Candida albicans: Estudo in vitro / Antifungal effect of different hydroalcoholic concentrations of Uncaria Tomentosa against Candida Albicans: In vitro study

En la actualidad el uso de la fitoterapia adquiere mayor demanda por parte de los pacientes y muchos profesionales de la salud debido a su fácil acceso, bajo costo y por presentar reacciones adversas mínimas. Objetivo: Determinar el efecto antifúngico de diferentes concentraciones hidroalcohólicas de la Uncaria tomentosa (UT) sobre Cándida albicans ATCC 10231, Mate-riales y métodos: Estudio experimental in vitro. La muestra estuvo constituida por 24 cajas petri con agar Sabourand, cada una con seis discos correspondientes a los 6 grupos de estudio siendo: G1 Nistatina de 21 ul (Control Positivo); G2 extracto hidroalcohólico de UT al 100%; G3 extracto hidroalcohólico de UT al 75%; G4 extracto hidroalcohólico de UT al 50%; G5 extracto hidroalcohólico de UT al 25%; G6 Alcohol etílico de 70° (Control negativo). Se elaboró un extracto hidroalcohólico por maceración, utilizando 75mg de planta micropulverizada y 250ml de alcohol etílico de 70°, se obtuvo 150ml de extracto, el mismo que fue diluido para obtener tres concentraciones secundarias al 75%, 50% y 25%. Los datos fueron procesados y analizados a través del test de ANOVA y de Bonferroni con un nivel de significancia de 5%. Resultados: Fue observado un halo de inhibición de 6.46 mm, 10.96 mm, 14.75 mm y 16,5 mm para los extractos hidroalcohólicos al 25%, 50%, 75% y 100% respectivamente. La Nistatina mostró un halo de inhibición de 23,42 mm y el alcohol etílico de 70° no obtuvo ningún efecto antifúngico. Fue observado diferencia estadísticamente significativa entre los grupos (< 0.001). Conclusión: El extrac-to hidroalcohólico al 100% de UT como la nistatina mostraron ser sensible, mientras que las concentraciones al 50% y 75% presentaron sensibilidad intermedia y la concentración al 25% fue resistente contra Candida albicans.

At present, the use of phytotherapy is in greater demand by patients and many health professionals due to its easy access, low cost and because of adverse reactions. Objective: To determine the antifungal effect of different hydroalcoholic concentrations of Uncaria tomentosa (UT) about Candida albicans ATCC 10231, Materials and methods: In vitro experimental study. The sample consisted of 24 box of petri dishes with Sabourand agar, each with six discs corresponding to the 6 study groups being: G1 Nistatin 21 ul (Positive Control); G2 hydroalcoholic extract of 100% UT; G3 hydroalcoholic extract of 75% UT; G4 hydroalcoholic extract of 50% UT; G5 hydroalcoholic extract of 25% UT; G6 70 ° Ethyl Alcohol (Negative Control) A hydroalcoholic extract was elaborated by maceration, using 75mg of micropulverized plant and 250ml of ethyl alcohol of 70 °, 150ml of extract was obtained, the same one that was diluted to obtain three secondary concentrations at 75%, 50% and 25%. The data were processed and analyzed through the ANOVA and Bonferroni test with a level of significance of 5%. Results: A halo of inhibition of 6.46 mm, 10.96 mm, 14.75 mm and 16.5 mm was observed for the hydroalcoholic extracts at 25%, 50%, 75% and 100% respectively. Nistatin showed a halo of inhibition of 23.42 mm and ethyl alcohol of 70 ° did not have any antifungal effect. A statistically significant difference was observed between the groups (<0.001). Conclusion: The 100% hydroalcoholic extract of UT such as nystatin showed to be sensitive, while the concentrations at 50% and 75% showed intermediate sensitivity and the concentration at 25% was resistant against Candida albicans.

Atualmente, o uso de fitoterapia está em maior demanda pelos pacientes e muitos profissionais da saúde devido ao seu fácil aces-so, baixo custo e por apresentar reações adversas mínimas. Objetivo: Determinar o efeito antifúngico de diferentes concentrações hidroalcoólicas de Uncaria tomentosa (UT) em Candida albicans ATCC 10231, Materiais e métodos: Estudo experimental in vitro. A amostra esteve constituída por 24 placas de Petri com agar Sabourand, cada uma com seis discos correspondentes aos 6 grupos de estudo: G1 Nistatina 21 ul (Controle Positivo); extracto hidroalcoólico G2 de 100% UT; extracto hidroalcoólico G3 de 75% UT; extracto hidroalcoólico G4 de 50% UT; extrato hidroalcoólico G5 de 25% UT; G6 álcool etílico a 70 ° (controle negativo). O extrato hidroalcoólico foi elaborado por maceração, utilizando 75 mg de planta micropulverizada e 250 ml de álcool etílico de 70 °, obteve-se 150 ml de extrato, o mesmo que foi diluído para obter três concentrações secundárias de 75%, 50% e 25%. Os dados foram processados e analisados através do teste ANOVA e Bonferroni com um nível de significância de 5%. Resultados: Observou-se um halo de inibição de 6,46 mm, 10,96 mm, 14,75 mm e 16,5 mm para os extratos hidroalcoólicos a 25%, 50%, 75% e 100%, respectivamente. A nistatina mostrou um halo de inibição de 23,42 mm e álcool etílico de 70 ° não teve nenhum efeito antifúngico. Observou-se diferença estatisticamente significativa entre os grupos (<0,001). Conclusão: O extracto hidroalcoólico 100% de UT assim como a nistatina mostraram ser sensíveis, enquanto as concentrações de 50% e 75% mostraram sensibilidade intermédia e a concentração de 25% foi resistente contra a Candida albicans.

Activity of the aqueous extract of Schinus terebinthifolius Raddi on strains of the Candida genus / Atividade do extrato aquoso de Schinus terebinthifolius Raddi sobre cepas do gênero Candida

ABSTRACT Objectives: To evaluate the antifungal susceptibility profile of the aqueous extract of the bark of Schinus terebinthifolius Raddi against the strains of the genus Candida. Methods: By using the disk diffusion method, 50 samples of the genus Candida (Candida albicans; Candida krusei; Candida glabrata; and Candida tropicalis), isolated from patients receiving treatment at Hospital Santa Casa de Misericórdia de São Paulo, and 1 American Type Culture Collection (ATCC) sample of each species were tested against: the isolated aqueous extract of the bark of Schinus terebinthifolius Raddi, isolated nystatin, and the association of nystatin and the aqueous extract of Schinus terebinthifolius Raddi. Results: There were no significant differences regarding the different strains of Candida tested. In the presence of the aqueous extract of Schinus terebinthifolius Raddi, no inhibition halo was visible. Isolated nystatin formed an inhibition halo measuring respectively 18.50 mm and 19.50 mm for the Candida albicans species and the others referred to as non-Candida albicans (Candida krusei; Candida glabrata; and Candida tropicalis). The association of nystatin and the aqueous extract of Schinus terebinthifolius Raddi resulted in inhibition halos measuring 14.25 mm and 16.50 mm respectively. The comparisons of these results are statistically significant (p < 0,001). Conclusion: The aqueous extract of Schinus terebinthifolius Raddi showed no antifun-gal activity in vitro against the strains tested, whereas the association of nystatin and the aqueous extract of Schinus terebinthifolius Raddi caused a decrease in the inhibition halo when compared with isolated nystatin.

RESUMO Objetivos: Avaliar o perfil de susceptibilidade antifúngica do extrato aquoso das cascas de Schinus terebinthifolius Raddi frente às cepas do gênero Candida. Métodos: Por meio do método de difusão em disco, 50 amostras do gênero Candida (Candida albicans, Candida krusei, Candida glabrata e Candida tropicalis) provenientes de pacientes do Hospital da Santa Casa de Misericórdia de São Paulo, e 1 amostra American Type Culture Collection (ATCC) de cada espécie foram testadas frente ao extrato aquoso das cascas de Schinus terebinthifolius Raddi isolado, nistatina isolada, e a associação da nistatina ao extrato aquoso de Schinus terebinthifolius Raddi. Resultados: Não houve diferenças significantes em relação às diferentes espécies de cepas de Candida testadas. O extrato aquoso de Schinus terebinthifolius Raddi não formou halo de inibição. A nistatina isolada formou halo de inibição de 18,50 mm e 19,50 mm respectivamente para as espécies Candida albicans e as demais nomeadas como não Candida albicans (Candida krusei, Candida glabrata e Candida tropicalis). A associação da nistatina ao extrato aquoso de Schinus terebinthifolius Raddi resultou no halo de inibição de 14,25 mm e 16,50 mm respectivamente, sendo que as comparações destes resultados são estatisticamente significantes (p < 0,001). Conclusão: O extrato aquoso de Schinus terebinthifolius Raddi não demonstrou propriedade antifúngica in vitro frente às cepas testadas, e a associação da nistatina ao extrato aquoso de Schinus terebinthifolius Raddi causou a diminuição do halo de inibição quando comparado à nistatina isolada.

Porosity of temporary denture soft liners containing antifungal agents

ABSTRACT Incorporation of antifungals in temporary denture soft liners has been recommended for denture stomatitis treatment; however, it may affect their properties. Objective: To evaluate the porosity of a tissue conditioner (Softone) and a temporary resilient liner (Trusoft) modified by minimum inhibitory concentrations (MICs) of antifungal agents for Candida albicans biofilm. Material and Methods: The porosity was measured by water absorption, based on exclusion of the plasticizer effect. Initially, it was determined by sorption isotherms that the adequate storage solution for specimens (65×10×3.3 mm) of both materials was 50% anhydrous calcium chloride (S50). Then, the porosity factor (PF) was calculated for the study groups (n=10) formed by specimens without (control) or with drug incorporation at MICs (nystatin: Ny-0.032 g, chlorhexidine diacetate: Chx-0.064 g, or ketoconazole: Ke-0.128 g each per gram of soft liner powder) after storage in distilled water or S50 for 24 h, seven and 14 d. Data were statistically analyzed by 4-way repeated measures ANOVA and Tukey's test (α=.05). Results: Ke resulted in no significant changes in PF for both liners in water over 14 days (p>0.05). Compared with the controls, Softone and Trusoft PFs were increased at 14-day water immersion only after addition of Ny and Chx, and Chx, respectively (p<0.05). Both materials showed no significant changes in PF in up to 14 days of S50 immersion, compared with the controls (p>0.05). In all experimental conditions, Softone and Trusoft PFs were significantly lower when immersed in S50 compared with distilled water (p<0.05). Conclusions: The addition of antifungals at MICs resulted in no harmful effects for the porosity of both temporary soft liners in different periods of water immersion, except for Chx and Ny in Softone and Chx in Trusoft at 14 days. No deleterious effect was observed for the porosity of both soft liners modified by the drugs at MICs over 14 days of S50 immersion.

Antifungal effects of synthetic human β-defensin 3-C15 peptide

OBJECTIVES: The purpose of this ex vivo study was to compare the antifungal activity of a synthetic peptide consisting of 15 amino acids at the C-terminus of human β-defensin 3 (HBD3-C15) with calcium hydroxide (CH) and Nystatin (Nys) against Candida albicans (C. albicans) biofilm. MATERIALS AND METHODS: C. albicans were grown on cover glass bottom dishes or human dentin disks for 48 hr, and then treated with HBD3-C15 (0, 12.5, 25, 50, 100, 150, 200, and 300 µg/mL), CH (100 µg/mL), and Nys (20 µg/mL) for 7 days at 37℃. On cover glass, live and dead cells in the biomass were measured by the FilmTracer Biofilm viability assay, and observed by confocal laser scanning microscopy (CLSM). On dentin, normal, diminished and ruptured cells were observed by field-emission scanning electron microscopy (FE-SEM). The results were subjected to a two-tailed t-test, a one way analysis variance and a post hoc test at a significance level of p = 0.05. RESULTS: C. albicans survival on dentin was inhibited by HBD3-C15 in a dose-dependent manner. There were fewer aggregations of C. albicans in the groups of Nys and HBD3-C15 (≥ 100 µg/mL). CLSM showed C. albicans survival was reduced by HBD3-C15 in a dose dependent manner. Nys and HBD3-C15 (≥ 100 µg/mL) showed significant fungicidal activity compared to CH group (p < 0.05). CONCLUSIONS: Synthetic HBD3-C15 peptide (≥ 100 µg/mL) and Nys exhibited significantly higher antifungal activity than CH against C. albicans by inhibiting cell survival and biofilm.

Lipid Raft Integrity Is Required for Survival of Triple Negative Breast Cancer Cells / 한국유방암학회지

PURPOSE: Lipid rafts are cholesterol enriched microdomains that colocalize signaling pathways involved in cell proliferation, metastasis, and angiogenesis. We examined the effect of methyl-β-cyclodextrin (MβCD)-mediated cholesterol extraction on the proliferation, adhesion, invasion, and angiogenesis of triple negative breast cancer (TNBC) cells. METHODS: We measured cholesterol and estimated cell toxicity. Detergent resistant membrane (DRM) and non-DRM fractions were separated using the OptiPrep gradient method. Cell cycles stages were analyzed by flow cytometry, apoptosis was assessed using the TdT-mediated dUTP nick end-labeling assay, and metastasis was determined using a Matrigel invasion assay. Neo-vessel pattern and levels of angiogenic modulators were determined using an in vitro angiogenesis assay and an angiogenesis array, respectively. RESULTS: The present study found that the cholesterol-depleting agent MβCD, efficiently depleted membrane cholesterol and caused concentration dependent (0.1–0.5 mM) cytotoxicity compared to nystatin and filipin III in TNBC cell lines, MDA-MB 231 and MDA-MB 468. A reduced proportion of caveolin-1 found in DRM fractions indicated a cholesterol extraction-induced disruption of lipid raft integrity. MβCD inhibited 52% of MDA-MB 231 cell adhesion on fibronectin and 56% of MDA-MB 468 cell adhesion on vitronectin, while invasiveness of these cells was decreased by 48% and 52% respectively, following MβCD treatment (48 hours). MβCD also caused cell cycle arrest at the G2M phase and apoptosis in MDA-MB 231 cells (25% and 58% cells, respectively) and in MDA-MB 468 cells (30% and 38% cells, respectively). We found that MβCD treated cells caused a 52% and 58% depletion of neovessel formation in both MDA-MB 231 and MDA-MB 468 cell lines, respectively. This study also demonstrated that MβCD treatment caused a respective 2.6- and 2.5-fold depletion of tyrosine protein kinase receptor (TEK) receptor tyrosine kinase levels in both TNBC cell lines. CONCLUSION: MβCD-induced cholesterol removal enhances alterations in lipid raft integrity, which reduces TNBC cell survival.

Obtenção de gel mucoadesivo de nistatina para o tratamento da candidíase oral. Desenvolvimento e caracterização de dispersões sólidas de nistatina / Mucoadhesive gel obtaining nystatin for the treatment of oral candidiasis. Development and characterization of solid dispersions of nystatin

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

Obtenção de gel mucoadesivo de nistatina para o tratamento da candidíase oral. Desenvolvimento e caracterização de dispersões sólidas de nistatina / Mucoadhesive gel obtaining nystatin for the treatment of oral candidiasis. Development and characterization of solid dispersions of nystatin

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

Efeito da incorporação de agentes antifúngicos na resistência à tração e porosidade de materiais resilientes temporários para base de próteses / Effect of incorporation of antifungal agents on the ultimate tensile strength and porosity of temporary soft denture liners

Este estudo investigou a resistência à tração (ou limite de resistência à tração- LRT) e a porosidade de reembasadores resilientes temporários modificados por concentrações inibitórias mínimas (CIMs) de agentes antifúngicos para o biofilme Candida albicans (SC5314). Para os testes de LRT, corpos de prova em forma de halteres (n=7) com uma área transversal de 33 mm x 6 mm x 3 mm foram produzidos para os materiais resilientes (Trusoft e Softone) sem (controle) ou com incorporação de cinco fármacos em suas CIMs: nistatina- 0,032 g; diacetato de clorexidina- 0,064; cetoconazol- 0,128 g; miconazol- 0,256 g; itraconazol-0,256 g (grama de fármaco por grama de pó de material resiliente). Após a plastificação, as amostras foram imersas em água destilada a 37°C durante 24 h, 7 e 14 dias e, então, testadas em tensão em uma máquina universal de ensaios (EMIC DL-500 MF) a 40 mm/min. A porosidade foi mensurada por absorção de água, com base na exclusão do efeito plastificante. Inicialmente, determinou-se por isotermas de sorção, que a solução de armazenagem adequada para os corpos de prova (65 mm x 10 mm x 3,3 mm) de ambos os materiais foi o cloreto de cálcio anidro a 50% (S50). Assim, o fator de porosidade (FP) foi calculado para os grupos de estudo (n=10) formados por espécimes sem (controle) ou com incorporação de fármaco em suas CIMs (nistatina, clorexidina ou cetoconazol) após a armazenagem em água destilada ou S50 por 24 h, 7 e 14 dias. Os dados de resistência à tração (MPa) e percentagem de alongamento (%) foram submetidos à ANOVA de 3 fatores seguida pelo teste de Tukey (=0,05). Os dados de porosidade foram analisados estatisticamente por ANOVA de medidas repetidas para 4 fatores e teste de Tukey (=0,05). Ao final de 14 dias, a resistência à tração para ambos os materiais foi significativamente menor nos grupos modificados pelo miconazol e itraconazol em relação aos outros grupos (P<0,0001), que não mostraram diferenças significativas entre si (P>0,05). Após 7 e 14 dias em água, o miconazol e itraconazol adicionados a ambos os materiais resultaram em percentagens significativamente menores de alongamento em comparação com os outros fármacos e ao controle (P<0,0001), que foram semelhantes entre si (P>0,05). O cetoconazol não resultou em alterações significativas no FP para ambos os materiais resilientes em água ao longo de 14 dias (P>0,05). Em comparação aos controles, houve aumento dos FPs do Softone e Trusoft aos 14 dias de imersão em água somente após a adição de nistatina e clorexidina e de clorexidina, respectivamente (P<0,05). Ambos os materiais não apresentaram alterações significativas no FP em até 14 dias de imersão na S50, em comparação aos controles (P>0,05). Em todas as condições experimentais, os FPs do Softone e Trusoft foram significativamente menores quando imersos em S50 em comparação com a água destilada (P<0,05). Concluiu-se que a adição de nistatina, clorexidina e cetoconazol nas CIMs para o biofilme de C. albicans não resultou em efeitos deletérios na resistência à tração e na percentagem de alongamento dos materiais resilientes temporários para base de prótese até o período de 14 dias. A adição de antifúngicos nas CIMs não resultou em efeitos adversos à porosidade de ambos os materiais resilientes temporários em diferentes períodos de imersão em água, com exceção da clorexidina e nistatina no Softone e clorexidina no Trusoft aos 14 dias. Não foram observados efeitos deletérios para a porosidade de ambos os materiais resilientes modificados com as CIMs dos fármacos durante os 14 dias de imersão na S50.(AU)

This study investigated the tensile strength (ultimate tensile strength- UTS) and porosity of temporary soft denture liners modified by minimum inhibitory concentrations (MICs) of antifungal agents for Candida albicans biofilm (SC5314). For UTS tests, dumbbell-shaped specimens (n=7) with a central cross-sectional area of 33 mm x 6 mm x 3 mm were produced by resilient materials (Trusoft and Softone) without (control) or with incorporation of five drugs at MICs: nystatin- 0.032 g; chlorhexidine diacetate-0.064 g; ketoconazole- 0.128 g; miconazole- 0.256 g; itraconazole- 0.256 g (each per gram of soft liner powder). After plasticization, specimens were immersed in distilled water at 37°C for 24 h, 7 and 14 days, and then tested in tension in a universal testing machine (EMIC DL-500 MF) at 40 mm/min. The porosity was measured by water absorption, based on exclusion of the plasticizer effect. Initially, it was determined by sorption isotherms that the adequate storage solution for specimens (65 mm x 10 mm x 3.3 mm) of both materials was 50% anhydrous calcium chloride (S50). Then, the porosity factor (PF) was calculated for the study groups (n=10) formed by specimens without (control) or with drug incorporation at MICs (nystatin, chlorhexidine or ketoconazole) after storage in distilled water or S50 for 24 h, 7 and 14 days. Data of tensile strength (MPa) and elongation percentage (%) were submitted to 3-way ANOVA followed by Tukey's test (=0.05). Data of porosity were statistically analyzed by 4-way repeated measures ANOVA and Tukeys test (=0.05). At the end of 14 days, the tensile strength for both materials was significantly lower in the groups modified by miconazole and itraconazole compared to the other groups (P<0.0001), which showed no significant difference between them (P>0.05). After 7 and 14 days in water, miconazole and itraconazole added into both materials result in significant lower elongation percentages compared to the other drugs and control (P<.0001), which were similar to each other (P>0.05). Ketoconazole resulted in no significant changes in PF for both liners in water over 14 days (P>0.05). Compared to the controls, Softone and Trusoft PFs were increased at 14-day water immersion only after addition of nystatin and chlorhexidine, and chlorhexidine, respectively (P<0.05). Both materials showed no significant changes in PF in up to 14 days of S50 immersion, compared to the controls (P>0.05). In all experimental conditions, Softone and Trusoft PFs were significantly lower when immersed in S50 compared to distilled water (P<0.05). It was concluded that the addition of the nystatin, chlorhexidine and ketoconazole at MICs for C. albicans biofilm resulted in no harmful effects on the ultimate tensile strength and elongation percentage of the temporary soft denture liners up to 14-day period. The addition of antifungals at MICs resulted in no detrimental effects for the porosity of both temporary soft liners in different periods of water immersion, except for chlorhexidine and nystatin in Softone and chlorhexidine in Trusoft at 14 days. No deleterious effect was observed for the porosity of both soft liners modified by the drugs at MICs over 14 days of S50 immersion.(AU)

Impact of brief exposure to antifungal agents on the post-antifungal effect and hemolysin activity of oral Candida albicans

AbstractPost-antifungal effect (PAFE) of Candida and its production of hemolysin are determinants of candidal pathogenicity. Candida albicans is the foremost aetiological agent of oral candidosis, which can be treated with polyene, azole, and echinocandin antifungals. However, once administered, the intraoral concentrations of these drugs tend to be subtherapeutic and transient due to the diluent effect of saliva and cleansing effect of the oral musculature. Hence, intra-orally, Candidamay undergo a brief exposure to antifungal drugs.Objective Therefore, the PAFE and hemolysin production of oral C. albicans isolates following brief exposure to sublethal concentrations of the foregoing antifungals were evaluated.Material and Methods A total of 50 C. albicans oral isolates obtained from smokers, diabetics, asthmatics using steroid inhalers, partial denture wearers and healthy individuals were exposed to sublethal concentrations of nystatin, amphotericin B, caspofungin, ketoconazole and fluconazole for 60 min. Thereafter, the drugs were removed and the PAFE and hemolysin production were determined by previously described turbidometric and plate assays, respectively.Results Nystatin, amphotericin B, caspofungin and ketoconazole induced mean PAFE (hours) of 2.2, 2.18, 2.2 and 0.62, respectively. Fluconazole failed to produce a PAFE. Hemolysin production of these isolates was suppressed with a percentage reduction of 12.27, 13.47, 13.33, 8.53 and 4.93 following exposure to nystatin, amphotericin B, caspofungin, ketoconazole and fluconazole, respectively.Conclusions Brief exposure to sublethal concentrations of antifungal drugs appears to exert an antifungal effect by interfering with the growth as well as hemolysin production of C. albicans.