Effectiveness of a tissue conditioner containing antifungals in a rat model of denture stomatitis.

AIM: This study investigated the effectiveness of a drug-modified tissue conditioner in an animal model of denture stomatitis. METHODS AND RESULTS: Wistar rats wore a Candida albicans-contaminated palatal device for 4 days. Next, nystatin (Nys) or chlorhexidine (Chx) were added to a tissue conditioner in their raw or ß-cyclodextrin-complexed (ßCD) forms at their minimum inhibitory concentrations. As controls, one group was not subjected to any procedure (NC), one group used sterile devices, one group had denture stomatitis but was not treated (DS), and another had the devices relined with the tissue conditioner without the addition of any drug (Soft). After 4 days of treatment, treatment effectiveness was assessed visually, histologically, and through CFU count, and myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) assays. Rats from the Soft, Nys, Nys:ßCD, and Chx groups presented a significant decrease in the microbial load compared with the untreated group. Treatment groups showed lower MPO and NAG activity compared to the non-treated group. CONCLUSIONS: The addition of antifungals to a soft tissue conditioner can be a promising approach for denture stomatitis treatment.

Anti-Candida activity and in vitro toxicity screening of antifungals complexed with ß-cyclodextrin.

The emergence of resistant fungal species and the toxicity of currently available antifungal drugs are relevant issues that require special consideration. Cyclodextrins inclusion complexes could optimize the antimicrobial activity of such drugs and create a controlled release system with few side effects. This study aimed to assess the in vitro toxicity and antifungal effectiveness of nystatin (Nys) and chlorhexidine (Chx) complexed or not with ß-cyclodextrin (ßCD). First, a drug toxicity screening was performed through the Artemia salina bioassay. Then, the minimum inhibitory concentrations (MICs) against Candida albicans were determined with the broth microdilution test. After MICs determination, the cytotoxicity of the drugs was evaluated through the methyl-thiazolyl-tetrazolium (MTT) and neutral red (NR) assays and through cell morphology analysis. The PROBIT analysis was used to determine the median lethal concentration (LC50), and the cell viability values were submitted to one-way analysis of variance(ANOVA)/Tukey (α = 0.05). Overall, the ßCD-complexed antifungals were less toxic against A. salina than their raw forms, suggesting that inclusion complexes can reduce the toxicity of drugs. The MICs obtained were as follows: Nys 0.5 mg/L; Nys:ßCD 4 mg/L; Chx 4 mg/L; and Chx:ßCD 8 mg/L. Chx showed significant cytotoxicity (MTT: 12.9 ± 9.6%; NR: 10.6 ± 12.5%) and promoted important morphological changes. Cells exposed to the other drugs showed viability above 70% with no cellular damage. These results suggest that antifungals complexed with ßCD might be a biocompatible option for the treatment of Candida-related infections.

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)

Antimicrobial photodynamic therapy in the treatment of oral erythematous candidiasis: a controlled and randomized clinical trial.

OBJECTIVE: To analyze the clinical and microbiological efficacy of antimicrobial photodynamic therapy (aPDT) in patients with erythematous candidiasis (EC). METHODS: This study was a controlled and randomized clinical trial in patients diagnosed with EC, who were allocated into a control group (CG) and experimental group (EG) treated with nystatin oral suspension and aPDT with methylene blue 0.1%, respectively. A clinical index was used to classify the EC lesions from mild to severe and assess the treatment efficacy. Microbiological samples were collected before and after aPDT session and analyzed by counting colony-forming units (CFUs) of Candida and Staphylococcus sp. RESULTS: A total of 41 patients (CG (n = 18); EG (n = 23)) were analyzed in our research. Of these, 16 (94.1%) of the CG and 16 (84.2%) of the EG exhibited complete remission of the lesions. Regarding the degree of the lesion, it was observed that the severe lesions were more difficult to present remission, while all the mild and moderate lesions showed complete regression (p = 0.001). The microbiological analysis showed that Candida albicans and Staphylococcus sp. were the most prevalent microorganisms, and the aPDT group showed a decrease in CFUs of these microorganisms after the first aPDT session (p < 0.05). CONCLUSIONS: aPDT proved to be a clinically and microbiologically effective therapy for treating EC. TRIAL REGISTRATION: Registered at ClinicalTrials.gov; Set 12th, 2019; No. RBR-8w8599. CLINICAL RELEVANCE: aPDT is a promising alternative treatment since it presents satisfactory results and does not cause damage to oral tissues or develop resistance to the treatment.

Assessing nystatin cream treatment efficacy against Leishmania (L.) amazonensis infection in BALB/c model.

The current scenario for cutaneous leishmaniasis treatment includes the use of first and second-choice drugs, both therapeutic strategies presenting several adverse effects and being related to an increment of treatment-refractory parasite strains. These facts encourage the search for new treatment approaches, including repositioning drugs, such as nystatin. Although in vitro assays show that this polyene macrolide compound has leishmanicidal activity, no in vivo evidence for a similar activity has been shown so far for the commercial nystatin cream formulation. This work assessed the effects of nystatin cream (25,000 IU/g) administered on mice in an amount to completely cover the paw surface of BALB/c mice infected with Leishmania (L.) amazonensis once a day, until a total of up to 20 doses. The data presented herein points to unequivocal evidence that treatment with this formulation causes a statistically significant reduction of swelling/edema in mice paws when compared to animal groups not submitted to this treatment regimen after the fourth week of infection: lesion sizes at the sixth (p = 0.0159), seventh (p = 0.0079) and eighth (p = 0.0079) week. Furthermore, swelling/edema reduction relates to a decrease in parasite load in the footpad (∼48%) and in draining lymph nodes (∼68%) at eight weeks post-infection. This is the first report of the effectiveness of nystatin cream used as a topical treatment in BALB/c model for cutaneous leishmaniasis.

Effectiveness of fluconazole as antifungal prophylaxis in cancer patients undergoing chemotherapy, radiotherapy, or immunotherapy: systematic review and meta-analysis.

This review assessed the effectiveness of fluconazole as antifungal prophylaxis on the incidence of oral fungal diseases in patients undergoing cancer treatment. The secondary outcomes evaluated were the adverse effects, discontinuation of cancer therapy due to oral fungal infection, mortality by a fungal infection, and the mean duration of antifungal prophylaxis. Twelve databases and records were searched. The RoB 2 and ROBINS I tools were used to assess the risk of bias. The relative risk (RR), risk difference, and standard mean difference (SMD) were applied with 95% confidence intervals (CI). The certainty of the evidence was determined by GRADE. Twenty-four studies were included in this systematic review. In randomized controlled trials pooling, fluconazole was a protective factor for the primary outcome (RR = 0.30; CI: 0.16, 0.55; p < 0.01, vs placebo). Compared to other antifungals, fluconazole was only more effective than the subgroup of amphotericin B and nystatin (alone or in combination) (RR = 0.19; CI: 0.09, 0.43; p < 0.01). Fluconazole was also a protective factor in non-randomized trials pooling (RR = 0.19; CI: 0.05, 0.78; p = 0.02, vs untreated). The results showed no significant differences for the secondary outcomes. The certainty of the evidence was low and very low. In conclusion, prophylactic antifungals are necessary during cancer treatment, and fluconazole was shown to be more effective in reducing oral fungal diseases only compared with the subgroup assessing amphotericin B and nystatin, administered alone or in combination.

Characterization, Antifungal Evaluation against Candida spp. Strains and Application of Nystatin: ß-cyclodextrin Inclusion Complexes.

BACKGROUND: Nystatin (Nys) is a fungicidal drug commonly prescribed for candidiasis disease in several administration routes. However, Nys is a class IV drug, according to the Biopharmaceutical Classification System, that possesses limited bioavailability and is used for local activity. OBJECTIVE: This study developed and characterized nystatin:ß-cyclodextrin (Nys:ßCD) inclusion complexes and evaluated their activity against Candida spp. METHODS: Complexes were characterized by physicochemical techniques and drug dissolution profiles. The susceptibility of C. albicans, C. krusei, C. parapsilosis, C. glabrata, C. guilliermondii, C. tropicalis, and C. auris was assessed using the broth microdilution method. The applicability of Nys:ßCD inclusion complex was evaluated by incorporating it into a temporary soft material for denture stomatitis treatment. RESULTS: Nys was better complexed in a 1:1 molar ratio by freeze-drying and spray-drying methods. The inclusion complexes show bi-exponential release, an initial burst release followed by a sustained manner, presenting higher dissolution efficiency than raw Nys. The 1:1 freeze-drying Nys:ßCD complex presents antifungal activity against all evaluated Candida strains, showing the maintenance of the drug effectiveness. The inclusion complex incorporated into a tissue conditioner material for denture stomatitis treatment effectively inhibited more than 90% of C. albicans biofilm growth during 7 and 14 days, in a half dose compared to raw Nys. CONCLUSION: This work represents a significant contribution to treating a wide variety of diseases caused by the Candida species, optimizing the drug bioavailability and compliance to the treatment due to improved drug solubility, dissolution, and sustained delivery.

Combined antibiofilm activity of synthetic peptides and antifungal drugs against Candida spp.

Introduction: Candida krusei and Candida albicans are biofilm-forming drug-resistant yeasts that cause bloodstream infections that can lead to death. Materials & methods: nystatin and itraconazole were combined with two synthetic peptides, PepGAT and PepKAA, to evaluate the synergistic effect against Candida biofilms. Additionally, scanning electron and fluorescence microscopies were employed to understand the mechanism behind the synergistic activity. Results: Peptides enhanced the action of drugs to inhibit the biofilm formation of C. krusei and C. albicans and the degradation of mature biofilms of C. krusei. In combination with antifungal drugs, peptides' mechanism of action involved cell wall and membrane damage and overproduction of reactive oxygen species. Additionally, in combination, the peptides reduced the toxicity of drugs to red blood cells. Conclusion: These results reveal that the synthetic peptides enhanced the antibiofilm activity of drugs, in addition to reducing their toxicity. Thus, these peptides have strong potential as adjuvants and to decrease the toxicity of drugs.

Candida krusei and Candida albicans are biofilm-forming, drug-resistant yeasts that cause bloodstream infections that can lead to death. In this study, biofilms of C. krusei and C. albicans were treated with a solution composed of synthetic peptides and antifungal drugs, none of which were effective alone. The synthetic peptides reduced the toxicity of drugs to red blood cells. These results may pave the way to the application of synthetic peptides as a beneficial additional to antifungal drugs to treat fungi that cannot be killed by drugs alone.

Nystatin effect on chlorhexidine efficacy against Streptococcus mutans as planktonic cells and mixed biofilm with Candida albicans.

OBJECTIVE: The aim of this study was to evaluate the effect of nystatin on the efficacy of chlorhexidine against Streptococcus mutans in planktonic cells and mixed biofilm with Candida albicans. MATERIAL AND METHODS: S. mutans ATCC 25,175 in suspension and also combined with C. albicans ATCC 18,804 in biofilm were cultured. Minimum inhibitory concentration (MIC), crystal violet colorimetric assay, and colony-forming unit counting (CFUs/mL) were performed. RESULTS: An increased MIC of chlorhexidine against S. mutans was observed when the drugs were administered mixed in a single formulation and with time intervals in between, except for the 30-min interval. The biofilm optical density (OD) in treatments using chlorhexidine and nystatin combined did not significantly differ from chlorhexidine alone. Either in biofilm colorimetric assay or determination of CFUs, the combined treatments with nystatin administered before chlorhexidine had less effect on chlorhexidine efficacy. CONCLUSIONS: Nystatin interferes with the action of chlorhexidine against S. mutans. The antimicrobial effectiveness of the combined drugs depends on their concentration, time interval used, and the planktonic or biofilm behavior of the microorganisms. CLINICAL RELEVANCE: In view of the great number of patients that can receive a prescription of chlorhexidine and nystatin concomitantly, this study contributes to the knowledge about the effect of the combined drugs. Given the high prevalence of prescriptions of chlorhexidine and nystatin in dentistry, dental professionals should be aware of their possible antagonistic effect.

Development of purified cashew gum mucoadhesive buccal tablets containing nystatin for treatment of oral candidiasis.

OBJECTIVE: The objective of this work was to prepare mucoadhesive buccal tablets containing nystatin and purified cashew gum for the treatment of oral candidiasis. SIGNIFICANCE: Mucoadhesive buccal tablets containing the drug nystatin are an alternative to oral suspensions, which cause low therapeutic adherence to the treatment of oral candidiasis. Purified cashew gum has been studied as a diluent and mucoadhesive agent in tablets. METHODS: Two batches of mucoadhesive tablets were produced, MT1 and MT 2, containing purified cashew gum, nystatin (500,000 IU), flavoring agent and with or without the presence of lubricant agent. The average weight, mechanical properties, dose uniformity, drug release profile, mucoadhesive properties and antimicrobial activity against Candida albicans were evaluated. RESULTS: Tablets presented average weight of 329.1 ± 3.1 mg (MT1) and 334.6 ± 1.5 mg (MT2), hardness of 9.8 ± 0.8 KgF (MT1) and 8.3 ± 0.4 KgF (MT2), friability of 0.2% (MT1 and MT2), and dose uniformity of 102.20 ± 1.17% (MT1) and 99.06 ± 7.40% (MT2). MT1 and MT2 were able to swell, erode, release the drug and remain adhered to the pig's cheek up to 3 h for batch MT1 and 4 h for batch MT2, and the amount of nystatin released since the beginning of the test in both batches was sufficient to inhibit the growth of the fungus. CONCLUSIONS: Therefore, the proposed formulation proved to be very promising and met all the studied criteria, showing to be ideal for the treatment of oral candidiasis.

Antifungal activity of Punicalagin-nystatin combinations against Candida albicans.

OBJECTIVES: Oral candidiasis is the most common opportunistic fungal infection of oral mucosa and results from an overgrowth of Candida, especially Candida albicans. The potential anti-C. albicans and cytotoxicity of punicalagin (PCG), isolated from Punica granatum, alone or with nystatin (NYS) were evaluated. METHODS: Activity of compounds alone or in combinations was determined against two C. albicans strains (ATCC 90028 and SC5314). Minimal inhibitory concentration (MIC)-50 and Minimum Fungicidal Concentration (MFC) were assessed by XTT assay and CFU counts, respectively. For combinations, determination of fractional inhibitory concentration index was performed. Ergosterol pathway was investigated as a possible PCG antifungal mechanism. Cytotoxicity assays were undertaken on human primary oral keratinocytes and gingival fibroblasts incubated with antifungal concentrations of PCG and/or NYS for 24 hr. RESULTS: Combination of NYS and PCG increased antifungal efficacy, compared with compounds tested alone. Combinations 4 (PCG-6.25 µg/ml; NYS-3.9 µg/ml) and 5 (PCG-12.5 µg/ml; NYS-1.95 µg/ml) were more effective since they reduced the MIC-50 of PCG (50 µg/ml) by 8 and 4 times, respectively, increased the candidal inhibition and nullified the PCG cytotoxicity for keratinocytes. PCG antifungal mechanism did not involve ergosterol biosynthesis pathway. CONCLUSIONS: The favorable outcomes for combination of PCG and NYS encourage further testing this therapeutic strategy against C. albicans.

Efficacy of copper sulphate on candida albicans on heat-polymerized acrylic resin / Eficacia del sulfato de cobre sobre candida albicans en resinas acrílicas de termocurado

The ageing of population is increasing, and a great percentage of these patients wear removable prostheses, and can suffer denture stomatitis, a condition that has been associated with candidiasis. Aims: To evaluate in vitro the effectiveness of Copper Sulfate against Candida albicans in samples of heat-polymerized acrylic resin, compared to nystatin, sodium hypochlorite and chlorhexidine. Materials and Methods: Initially, the minimum inhibitory concentration (MIC) of copper sulfate for Candida albicans was determined by microdilution. Then, 54 resin samples were divided into 6 treatment groups corresponding to Nystatin 100.000 UI, Sodium Hypochlorite 0.5%, chlorhexidine 0.12%, Copper Sulfate 4.7µg/ml, Copper Sulfate 9.4µg/ml and physiological saline solution, in which samples were submerged for 6 hours. Resin samples were then washed and cultured on solid media at 37°C for 72 hours. The number of colony-forming units was determined using a Quebec colony counter. The statistical analysis was performed using the Kruskal-Wallis test and the Mann-Whitney U test. Results: Copper sulfate at a concentration of 9.4µg/ml presented a similar effectiveness as the other control products regarding the reduction in the number of colonies of Candida albicans post-treatment. Conclusion: The effectiveness of copper sulfate against Candida albicans on acrylic resin was similar to that of nystatin, sodium hypochlorite and chlorhexidine.

En las últimas décadas se ha observado un aumento de la población de adultos mayores, de los cuales un gran porcentaje es portador de prótesis removible, y dos tercios pueden sufrir estomatitis subprotésica, enfermedad que es asociada a infecciones como candidiasis. Objetivo: Evaluar la efectividad antimicótica in vitro del sulfato de cobre en placas de resinas acrílicas de termocurado inoculadas con Candida albicans, frente a Nistatina, Hipoclorito de Sodio y Clorhexidina. Material y Métodos: Inicialmente, y mediante microdilución del sulfato de cobre, se determinó la concentración mínima inhibitoria (CMI) para Candida albicans. En la fase experimental, 54 muestras de resina se dividieron en 6 grupos correspondientes a Nistatina 100.000 UI, Hipoclorito 0.5%, Clorhexidina 0.12%, Sulfato de Cu 4.7µg/ml, Sufato de Cu 9.4 µg/ml y suero fisiológico. Las muestras fueron sumergidas en estos agentes por 6 horas, para posteriormente ser lavadas y cultivada en medios solidos a 37°C por 72 horas. Luego se realizó el conteo de unidades formadoras de colonias mediante contador tipo Quebec. El análisis estadístico se realizó mediante la prueba de Kruskal-Wallis y la prueba U de Mann-Whitney. Resultado: El sulfato de cobre a una concentración de 9.4µg/ ml presentó una efectividad similar a los otros productos, en la reducción de colonias de Candida albicans. Conclusión: La efectividad del sulfato de cobre contra Candida albicans fue semejante a la de Nistatina, Hipoclorito y Clorhexidina.

Uso da terapia fotodinâmica antimicrobiana no tratamento de candidose oral: ensaio clínico, controlado e randomizado / Use of antimicrobial photodynamic therapy in the treatment of oral candidosis: a randomized, controlled clinical trial

INTRODUÇÃO: A candidose eritematosa oral é uma infecção fúngica comum em usuários de prótese. Em algumas situações os pacientes exibem resistência ao tratamento convencional. Novas terapias vêm surgindo como alternativas para o seu tratamento, como a terapia fotodinâmica antimicrobiana (TFDa). OBJETIVO: Analisar a eficácia da TDFa no tratamento da candidose eritematosa oral em comparação com o tratamento convencional a base de nistatina. METODOLOGIA: O presente estudo consiste em um ensaio clínico, controlado e randomizado, realizado por meio de um estudo clínico e análise microbiológica em pacientes diagnosticados com candidose eritematosa oral, os quais foram divididos entre grupo controle, tratados com suspensão oral de nistatina e grupo caso, tratados com sessões de TFDa. Os pacientes foram avaliados quanto a eficácia do tratamento, através da aplicação de um índice clínico, além de serem realizadas análises microbiológicas, em períodos préestabelecidos (7,14,21 e 30 dias). RESULTADOS: Dos 41 pacientes analisados, sendo 18 deles do grupo controle e 23 do grupo caso, 32 (78%) obtiveram sucesso dos tratamentos utilizados. Quanto ao grau clínico, as severas exibiram mais dificuldade em apresentar remissão, enquanto todas as lesões leves e moderadas regrediram. O grupo caso, exibiu resultados semelhantes ao do grupo controle, no entanto, não houve diferenças estatisticamente significativa entre os grupos, tanto quanto ao sucesso do tratamento, nas análises microbiológicas, no qual foi contatado o predomínio da Candida albicans,entre as espécies de Candida ssp. e de Staphylococcus ssp. nas próteses dentarias. CONCLUSÕES: A TFDa é uma alternativa de tratamento promissora, por apresentar resultados satisfatórios, não causar alterações aos tecidos orais e seu uso não levar ao desenvolvimento de resistência ao tratamento (AU).

INTRODUCTION: Oral erythematous candidiasis is a common fungal infection in users of complete dentures. patients often exhibit resistance to conventional treatment. New therapies are emerging as alternatives for its treatment, such as antimicrobial photodynamic therapy(aPDT). AIM:Analysis in patients diagnosed with oral erythematous candidiasis, who were divided into a control group, treated with oral nystatin suspension, and an intervention group, treated with aPDT sections. METHODS: The present study consists of a clinical, controlled and randomized trial conducted by means of a clinical and microbiological analysis in patients diagnosed with oral erythematous candidiasis, who were divided into a control group, treated with oral nystatin suspension, and an intervention group, treated with aPDT sections. Patients were reassessed as to the effectiveness of treatment through the application of a clinical index, in addition to microbiological analysis,in pre-established periods (7,14,21 and 30 days).RESULTS:Of 41 patients analyzed in our research, 32 (78%) were successful in their treatment,4 (9,8%) were unsuccessful, and 5 (12,2%) gave up treatment. Patients were also analyzed for the degree of the lesion, and it was observed that those of the severe type had more difficulty in presenting remission, while the lesions mild and moderate, all showed regression. The case group exhibited similar results to the control group, however, there were no statistically significant differences between the groups, as far as treatment success was concerned, in microbiological analyzes, in which the predominance of Candida albicans, among the species of Candida ssp. and Staphylococcus ssp. in dental prostheses. CONCLUSIONS: aPDT is a promising treatment alternative, as it presents satisfactory results, does not cause changes to oral tissues and its use does not lead to the development of resistance to treatment, so further studies with larger samples are needed to verify its effectiveness (AU).

Atividade antifúngica do a-terpinen sobre Candida albicans / Antifungal activity of a-terpinen on Candida albicans

Objetivo: avaliar a atividade antifúngica do a- terpinen sobre culturas planctonicas e biofilme de Candida albicans. Material e Métodos: Primeiramente, foi determinada a Concentração Inibitória Mínima (CIM) e a Concentração Fungicida Mínima (CFM) do a-terpinen sobre microrganismos planctônicos. A Nistatina foi utilizada como controle positivo. Biofilme de Candida albicans foi desenvolvido e, após o tratamento com diferentes concentrações de α-terpinen, foi quantificado em UFC/mL, além da atividade metabólica das células ser avaliada por XTT. Resultados: a menor concentração capaz de inibir o crescimento (CIM) foi 0,2 % para o a-terpinen e 4 µg/mL para a Nistatina. Na CIM, os resultados mostraram que a partir da concentração 0,05 % de α-terpinen e 2 µg/mL de Nistatina houve diminuição de C.albicans quando comparado ao controle. A CFM foi para a-terpinen 0,2 % e Nistatina 8 µg/mL. Na quantificação as concentrações eficazes foram de α-terpinen (0,1%) e Nistatina (128µg/mL), e no teste do XTT, observou-se que α -terpinen (0,1%) e Nistatina (256µg/mL) diminuem a viabilidade quando comparado com o controle. Conclusão: Assim, pode-se afirmar que α -terpineol pode ser uma alternativa para tratamento de infecções fúngicas.

Objective: to evaluate the antifungal activity of a-terpinen on planktonic cultures and biofilm of Candida albicans. Material and Methods: first, Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (CFM) of a-terpinen were determined. Nystatin was used as a positive control. Biofilm of Candida albicans was developed and, after treatment with different concentrations of a-terpinen, was quantified in CFU/mL, in addition to metabolic activity of the cells being evaluated by XTT. Results: the lowest concentration able to inhibit the growth (MIC) was 0.2% for a-terpinen and 4 µg / mL for Nystatin. Results showed that from the concentration 0.05% of α -terpinen and 2 µg / mL of Nystatin, there was a decrease of Candida albicans when compared to the control, in planktonic culture. CFM was 0.2% for α -terpinen and 8 µg / mL for Nystatin. Regarding the quantification, effective concentrations were α-terpinen (0.1%) and Nystatin (128 µg/mL), and in the XTT test, α-terpinen (0.1%) and Nystatin (256 µg/mL) decreased metabolic activity when compared to control. Conclusion: Thus, it can be stated that a-terpineol may be an alternative for the treatment of fungal infections.

Activity of Fenticonazole, Tioconazole and Nystatin on New World Leishmania Species.

Leishmaniasis is an infectious disease caused by protozoal parasites belonging to Leishmania genus. Different clinical outcomes can be observed depending on the parasite species and health condition of patients. It can range from single cutaneous lesion until deadly visceral form. The treatment of all forms of leishmaniasis is based on pentavalent antimonials, and in some cases, the second-line drug, amphotericin B is used. Beside the toxicity of both drugs, parasites can be resistant to antimonial in some areas of the world. This makes fundamental the characterization of new drugs with leishmanicidal effect. Thus, the aim of the present work was to study the leishmanicidal activity of drugs able to interfere with ergosterol pathway (fenticonazole, tioconazole, nystatin, rosuvastatin and voriconazole) against promastigote and amastigote forms of L.(L.) amazonensis, L.(V.) braziliensis and L.(L.) infantum, and its impact on morphological and physiological changes in L.(L.) amazonensis or in host macrophages. We observed that fenticonazole, tioconazole and nystatin drugs eliminated promastigote and intracellular amastigotes, being fenticonazole and nystatin the most selective towards amastigote forms. Rosuvastatin and voriconazole did not present activity against amastigote forms of Leishmania sp. In addition, the drugs with leishmanicidal activity interfered with parasite mitochondrion. Although drugs did not stimulate NO and H2O2, specially fenticonazole was able to alkalize infected host macrophages. These results suggest well established and non-toxic antifungal drugs can be repurposed and used in leishmaniasis.

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.

Diversity, frequency and antifungal resistance of Candida species in patients with type 2 diabetes mellitus.

OBJECTIVE: To determine number, species of Candida and Candida resistance to antifungal therapy according to the metabolic control state and the associated salivary changes in patients with type 2 diabetes mellitus (DM2). MATERIALS AND METHODS: Samples of non-stimulated saliva were collected from 52 patients with DM2. Salivary pH was measured and cultured on Sabouraud glucose agar and the values of CFU/ml were calculated. The species were presumptively identified using CHROMagar Candida® plates, and identification was confirmed by polymerase chain reaction (PCR). C. albicans isolates were cultured on SGA tetracycline agar with nystatin and fluconazole diffusion disks to measure susceptibility. RESULTS: Sixty six percent of the yeasts isolated were Candida albicans, followed by C. glabrata (20.7%). In patients with decompensated DM2, there was an inverse association between HbA1c value and salivary pH. At higher levels of salivary acidification, a greater diversity and quantity of yeasts of the genus Candida were observed. With nystatin, higher inhibition was observed at lower pH. CONCLUSIONS: The antifungal therapies could be more effective if it consider, qualitative salivary characteristics as pH, that could determine the susceptibility of species of Candida to at least to nystatin, which is the most used antifungal for treatment to oral candidiasis in patients with DM2.

Stability and efficacy of combined nystatin and chlorhexidine against suspensions and biofilms of Candida albicans.

OBJECTIVE: Nystatin and chlorhexidine are extensively used in oral medicine; however, there is some controversy about the possibility of these drugs showing antagonism. To clarify this issue, this study investigated the efficacy and stability of nystatin and chlorhexidine in combination. DESIGN: An in vitro study was conducted to analyze the effect of nystatin and chlorhexidine combined on Candida albicans ATCC 18804, using the drugs mixed as a single formulation and as independent formulations used sequentially with different time intervals between them. The minimum inhibitory concentration (MIC) and effects on C. albicans suspensions and biofilms were evaluated. Also, the stability of nystatin and chlorhexidine in a mixture was tested by high performance liquid chromatography (HPLC). RESULTS: When nystatin and chlorhexidine were mixed in a single formulation, there was no significant difference in MIC compared to that of the drugs used alone (as the only treatment). However, when these drugs were used as independent formulations, sequentially with time intervals in between, their MICs were higher than the respective MIC of the drug used alone, except for the MIC of chlorhexidine with a 10-min interval. Nystatin/chlorhexidine combinations showed lower activity against C. albicans biofilms, except for that with a 30-min interval. The drugs when combined showed high percentages of degradation at all the times evaluated. CONCLUSIONS: The combination of nystatin and chlorhexidine seems to interfere with the efficacy of the drugs and to increase their rate of degradation.

Use of nystatin and chlorhexidine in oral medicine: Properties, indications and pitfalls with focus on geriatric patients.

OBJECTIVES: The aim of this work was to review the scientific literature on the properties, indications and pitfalls related to nystatin and chlorhexidine in oral medicine and also to compare these to other topical antifungal agents, considering the elderly population. BACKGROUND: Nystatin is a polyene antifungal widely used as a topical formulation to treat candidiasis, whereas chlorhexidine is a wide-spectrum antimicrobial, especially used against bacteria, but also effective in treating some fungal infections including those caused by Candida spp. These compounds have been prescribed for immunocompromised patients, hospitalized or not, some of them undergoing head and neck radiation therapy and/or chemotherapy, including elderly patients. MATERIALS AND METHODS: Dental and medical literature concerning the use of nystatin and chlorhexidine in oral medicine were selected and reviewed. RESULTS: Nystatin and chlorhexidine are gold-standard antimicrobial mouthrinses respectively for Candida spp. and bacteria. Although recognized as effective in cotrolling oral infections, both nystatin and chlorhexidine are just complementary to systemic therapy in cases of systemic infections already established. The prescriber should also take into account that some commercial nystatin and chlorhexidine formulations contain compounds such as sugar and ethanol, which can be associated with side effects. Meanwhile, alternative formulations in which these compounds are absent are available and should be considered. CONCLUSIONS: Further studies investigating new drugs and interactions of drug combinations are necessary to improve the therapeutic management of oral infections.