RESUMO
Introduction. Tissue conditioners modified with antifungals are a potential alternative to denture stomatitis (DS) treatment.Gap Statement. Information on tissue response to this treatment before its clinical application is lacking.Aim. This study aimed to evaluate the tissue response of a tissue conditioner modified with antifungals in a rat model of DS.Methodology. After DS induction for 4 days under antibiotic therapy, Wistar rats had their intraoral devices (IODs) relined with the tissue conditioner Softone without (Soft) or with the MICs against Candida albicans of nystatin (Nys) or chlorhexidine (Chx) complexed or not with ß-cyclodextrin (Nys:ßCD and Chx:ßCD). Three controls were included: healthy rats [negative control (Nc)], rats using a sterile IOD [sterile device (Sd)] and rats with DS that did not receive treatment (DS). After 4 days of treatment, the palatal mucosa under the IODs underwent histological processing for morphohistopathological and histometric analyses, morphology of collagen fibres (birefringence), immunohistochemistry for the expression of cell proliferation (proliferating cell nuclear antigen) and cytokine (IL-1ß).Results. The Nc and Sd groups were similar (P>0.05), displaying epithelial and connective tissues without any discernible changes in the parameters assessed. The DS and Soft groups exhibited pronounced epithelial alterations, cell proliferation and expression of the cytokine IL-1ß. In groups treated with drug incorporation (Nys, Chx, Nys:ßCD and Chx:ßCD), all samples demonstrated a reduction in tissue inflammation or complete tissue recovery, with an epithelium compatible with health. For the immunohistochemical parameters, the Chx, Nys:ßCD and Chx:ßCD groups were comparable with Nc (P>0.05).Conclusion. The proposed treatment could be promising for DS, as it led to the tissue recovery of the palatal mucosa. Nevertheless, much lower concentrations of complexed antifungals were required to achieve a similar or higher degree of tissue response compared with uncomplexed drugs in a modified tissue conditioner formulation.
Assuntos
Antifúngicos , Candida albicans , Modelos Animais de Doenças , Mucosa Bucal , Nistatina , Ratos Wistar , Estomatite sob Prótese , beta-Ciclodextrinas , Animais , beta-Ciclodextrinas/química , Antifúngicos/farmacologia , Estomatite sob Prótese/tratamento farmacológico , Estomatite sob Prótese/microbiologia , Ratos , Nistatina/farmacologia , Nistatina/administração & dosagem , Candida albicans/efeitos dos fármacos , Mucosa Bucal/efeitos dos fármacos , Mucosa Bucal/microbiologia , Masculino , Clorexidina/farmacologia , Interleucina-1beta/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Testes de Sensibilidade MicrobianaRESUMO
Patients with orofacial clefts are more likely to develop oral fungal diseases due to anatomo-physiological changes and surgical rehabilitation treatment. This case-series study evaluated the genetic diversity and dynamics of oral colonization and spread of C. albicans and C. tropicalis in four patients with orofacial clefts, from the time of hospital admission, perioperative and outpatient follow-up, with specialized physician. Candida biotypes previously identified by CHROMagar Candida and PCR methods were studied by MALDI-TOF MS assays and clustering analyses. Possible correlations with pathogenicity characteristics were observed, including production of hydrolytic exoenzymes and the antifungal sensitivity profiles. Amphotericin B-sensitive and fluconazole-resistant (low frequency) C. tropicalis and C. albicans, including clinically compatible MIC of nystatin, were found in the oral cavity of these patients. Clusters of isolates revealed phenomena of (i) elimination in the operative phase, (ii) maintenance or (iii) acquisition of oral C. tropicalis in the perioperative period and specialized outpatient and medical follow-up. For C. albicans, these phenomena included (i) elimination in the operative phase, (ii) acquisition in the operative phase and propagation from the hospital environment, and (iii) maintenance during hospitalization and operative phase. Amphotericin B and nystatin were shown to be effective in cases of clinical treatment and/or prophylaxis, especially considering the pre-existence of fluconazole-resistant strains. This study confirmed the phenomena of septic maintenance, septic neocolonization and septic elimination involving the opportunistic pathogens. MALDI-TOF MS associated with clustering analysis may assist the monitoring of clinical isolates or groups of epidemiologically important microbial strains in the hospital setting.
Assuntos
Anfotericina B , Antifúngicos , Candida albicans , Candida tropicalis , Farmacorresistência Fúngica , Genótipo , Testes de Sensibilidade Microbiana , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Humanos , Candida tropicalis/efeitos dos fármacos , Candida tropicalis/isolamento & purificação , Candida tropicalis/genética , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candida albicans/genética , Candida albicans/efeitos dos fármacos , Candida albicans/isolamento & purificação , Anfotericina B/farmacologia , Anfotericina B/uso terapêutico , Masculino , Feminino , Boca/microbiologia , Criança , Fluconazol/farmacologia , Fluconazol/uso terapêutico , Candidíase Bucal/microbiologia , Nistatina/farmacologia , Nistatina/uso terapêutico , Técnicas de Tipagem Micológica , Fissura Palatina/cirurgia , Fenda Labial/cirurgia , Adolescente , Análise por Conglomerados , Pré-EscolarRESUMO
Although Streptococcus pyogenes and Candida albicans may colonize tonsillar tissues, the interaction between them in mixed biofilms has been poorly explored. This study established an interkingdom biofilm model of S. pyogenes and C. albicans and verified the dose-response validation of antimicrobials. Biofilms were formed on microplates, in the presence or absence of a conditioning layer of human saliva, using Brain Heart Infusion (BHI) broth or artificial saliva (AS) as a culture medium, and with variations in the microorganism inoculation sequence. Biofilms grown in AS showed higher mass than those grown in BHI broth, and an opposite trend was observed for metabolism. The number of S. pyogenes colonies was lower in AS. Amoxicillin and nystatin showed dose-dependent effects. The inoculation of the two species at the same time, without prior exposure to saliva, and using BHI broth would be the model of choice for future studies assessing the effects of antimicrobials on dual S. pyogenes/C. albicans biofilms.
Assuntos
Biofilmes , Candida albicans , Streptococcus pyogenes , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Biofilmes/efeitos dos fármacos , Streptococcus pyogenes/efeitos dos fármacos , Streptococcus pyogenes/fisiologia , Humanos , Relação Dose-Resposta a Droga , Saliva/microbiologia , Testes de Sensibilidade Microbiana , Meios de Cultura/química , Amoxicilina/farmacologia , Nistatina/farmacologia , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologiaRESUMO
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.
Assuntos
Antifúngicos , Candida albicans , Clorexidina , Nistatina , Ratos Wistar , Estomatite sob Prótese , Animais , Estomatite sob Prótese/microbiologia , Estomatite sob Prótese/tratamento farmacológico , Ratos , Antifúngicos/uso terapêutico , Antifúngicos/farmacologia , Nistatina/farmacologia , Nistatina/uso terapêutico , Clorexidina/farmacologia , Candida albicans/efeitos dos fármacos , Modelos Animais de Doenças , Masculino , Contagem de Colônia Microbiana , Testes de Sensibilidade Microbiana , Candidíase Bucal/tratamento farmacológico , Candidíase Bucal/microbiologia , Peroxidase/metabolismo , Acetilglucosaminidase/metabolismo , beta-CiclodextrinasRESUMO
This study aimed to answer the question formulated according to the PICO strategy: 'Which essential oils show antimicrobial activity against biofilms formed on dental acrylic resin?' composed by population (dental acrylic resin), intervention (application of essential oils), comparison (denture cleansers, antifungal drugs, chlorhexidine, and oral mouthwashes), and outcome (antibiofilm activity). In vitro experimental studies evaluating the activity of EOs on biofilm formed on acrylic resin were included. PRISMA guidelines were followed, and the search was performed in the PubMed, Science Direct, Embase, and Lilacs databases and in the gray literature using Google Scholar and ProQuest in December 2023. A manual search of the reference lists of the included primary studies was performed. Of the 1467 articles identified, 37 were selected for full-text reading and 12 were included. Twelve EOs were evaluated, of which 11 showed activity against Candida spp., 3 against Staphylococcus aureus, and 1 against Pseudomonas aeruginosa. The EOs of Cymbopogon citratus, Cinnamomum zeylanicum, and Cymbopogon nardus showed higher action than chlorhexidine, C. nardus higher than Listerine, C. citratus higher than nystatin, and Melaleuca alternifolia higher than fluconazole and nystatin. However, chlorhexidine was more effective than Lippia sidoides and Salvia officinalis, sodium hypochlorite was more effective than L. sidoides, nystatin was more effective than Zingiber officinale, Amphotericin B more effective than Eucalyptus globulus and M. alternifolia. In conclusion, the EOs of C. zeylanicum, C. citratus, C. nardus, and M. alternifolia showed antimicrobial activity to reduce biofilm on dental acrylic resin.
Assuntos
Resinas Acrílicas , Biofilmes , Óleos Voláteis , Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans , Clorexidina/farmacologia , Nistatina/farmacologia , Óleos Voláteis/farmacologiaRESUMO
The pathogenic nature of infections caused by Candida spp. underscores the necessity for novel therapeutic agents. Extracts of Schinopsis brasilienses Engl are \ a promising source of agents with antifungal effects. This study aimed to assess the antifungal potential of the leaf extract of S. brasilienses. The antifungal activity was evaluated by determining the minimum inhibitory concentrations and fungicide concentrations (MIC and MFC). The antibiofilm potential was assessed by counting colony-forming units/mL. The study examined the inhibition kinetics of fungal growth and potential synergism between gallic acid or the extract and nystatin using the Checkerboard method. Cytotoxicity was evaluated through the MTT assay. The extract exhibited antifungal effect against all tested strains, with MIC and MFC ranging from 31.25-250 µg/mL. Gallic acid, the main isolated compound, displayed a MIC of 2000 µg/mL. The extract of S. brasilienses at 31.25 µg/mL inhibited the formation of biofilm by C. albicans and significantly reduced the mass of mature biofilm after 24 and 48 h (p < 0. 05). At a concentration of 125 µg/mL, the extract demonstrated significant inhibition of fungal growth after 6 hours. The combination of gallic acid or extract with nystatin did not exhibit synergistic or antagonistic effect. Furthermore, the extract did not induce cytotoxicity to a human cell line. The extract of S. brasiliensis demonstrates antifungal activity against Candida, generally exhibiting fungicidal action and capacity to inhibit biofilm formation as well as reduce mature biofilms. Additionally, the extract showed low cytotoxicity to human cells.
Assuntos
Anacardiaceae , Candida , Humanos , Antifúngicos , Nistatina , Candida albicans , Biofilmes , Ácido Gálico , Extratos VegetaisRESUMO
OBJECTIVE: This study proposed to assess the effect of Cryptocarya moschata extract on single and mixed biofilms formed on denture base and reline acrylic resin. MATERIALS AND METHODS: Single and mixed biofilms of Candida albicans and Streptococcus mutans were formed on the samples and treated with C. moschata extract; Nystatin solution at 100,000 IU/mL or Penicillin antibiotic solution at 100,000 IU/mL; or PBS solution. Antimicrobial activity was analyzed by counting colony-forming units, metabolism assay, assessment of protein components of the biofilm matrix, and of cell viability using confocal laser scanning microscopy (CLSM). Data were submitted to ANOVA and Tukey's post-test (α = 0.05). RESULTS: Cryptocarya moschata extract reduced cell viability of C. albicans and S. mutans single and mixed biofilms formed on samples. For all types of biofilms in the C. moschata group, there was a log reduction of the biofilm, proven by the Alamar Blue assay. Analyzing the extracellular matrix protein components, groups treated with the extract exhibited a lower level of fluorescence compared to the PBS groups. Reduction in thickness biofilm and viable cells was perceptible in the C. moschata group when assessing through CLSM. CONCLUSION: Cryptocarya moschata extract reduced the single and mixed biofilms of C. albicans and S. mutans on acrylic resins.
Assuntos
Resinas Acrílicas , Biofilmes , Candida albicans , Bases de Dentadura , Extratos Vegetais , Streptococcus mutans , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Resinas Acrílicas/farmacologia , Streptococcus mutans/efeitos dos fármacos , Extratos Vegetais/farmacologia , Bases de Dentadura/microbiologia , Microscopia Confocal , Nistatina/farmacologiaRESUMO
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.
Assuntos
Antifúngicos , beta-Ciclodextrinas , Antifúngicos/toxicidade , Candida , Nistatina/toxicidade , Candida albicans , Clorexidina/farmacologia , beta-Ciclodextrinas/toxicidadeRESUMO
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)
Assuntos
Estomatite sob Prótese , Candida albicans , NistatinaRESUMO
Aim: This study evaluated the antifungal efficacy of gentian violet (GV) in an experimental vulvovaginal candidiasis (VVC) model. Materials & methods: In vitro susceptibility and cytotoxicity assays were performed to validate the antifungal potential and safety of GV. The antifungal efficacy was then evaluated in vivo through comparative analysis of the fungal burden following treatment with GV or nystatin, as well as assessment of the vaginal tissue by histology and electron microscopy. Results: GV demonstrated a safe antifungal profile against C. albicans, with a significant decrease in fungal burden and an improvement in the inflammatory process evaluated histologically. Conclusion: The results of this study motivate further assessment of GV as a promising alternative for VVC therapy.
Assuntos
Candidíase Vulvovaginal , Feminino , Humanos , Camundongos , Animais , Candidíase Vulvovaginal/tratamento farmacológico , Candidíase Vulvovaginal/microbiologia , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Violeta Genciana/uso terapêutico , Candida albicans , Nistatina/farmacologia , Nistatina/uso terapêuticoRESUMO
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 (â¼48%) and in draining lymph nodes (â¼68%) 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.
Assuntos
Leishmania , Leishmaniose Cutânea , Animais , Camundongos , Nistatina/farmacologia , Nistatina/uso terapêutico , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/parasitologia , Resultado do Tratamento , Edema , Camundongos Endogâmicos BALB CRESUMO
We developed a simple new selective LB-based medium, named CYP broth, suitable for recovering long-term stored Y. pestis subcultures and for isolation of Y. pestis strains from field-caught samples for the Plague surveillance. It aimed to inhibit the growth contaminating microorganisms and enrich Y. pestis growth through iron supplementation. The performance of CYP broth on microbial growth from different gram-negative and gram-positive strains from American Type Culture Collection (ATCC®) and other clinical isolates, field-caught rodent samples, and more importantly, on several vials of ancient Y. pestis subcultures was evaluated. Additionally, other pathogenic Yersinia species such as Y. pseudotuberculosis and Y. enterocolitica were also successfully isolated with CYP broth. Selectivity tests and bacterial growth performance on CYP broth (LB broth supplemented with Cefsulodine, Irgasan, Novobiocin, nystatin and ferrioxamine E) were evaluated in comparison with LB broth without additive; LB broth/CIN, LB broth/nystatin and with traditional agar media including LB agar without additive, and LB agar and Cefsulodin-Irgasan-Novobiocin Agar (CIN agar) supplemented with 50 µg/mL of nystatin. Of note, the CYP broth had a recovery twofold higher than those of the CIN supplemented media or other regular media. Additionally, selectivity tests and bacterial growth performance were also evaluated on CYP broth in the absence of ferrioxamine E. The cultures were incubated at 28 °C and visually inspected for microbiological growth analysis and O.D.625 nm measurement between 0 and 120 h. The presence and purity of Y. pestis growth were confirmed by bacteriophage and multiplex PCR tests. Altogether, CYP broth provides an enhanced growth of Y. pestis at 28 °C, while inhibiting contaminant microorganisms. The media is a simple, but powerful tool to improve the reactivation and decontamination of ancient Y. pestis culture collections and for the isolation of Y. pestis strains for the Plague surveillance from various backgrounds. KEY POINTS: ⢠The newly described CYP broth improves the recuperation of ancient/contaminated Yersinia pestis culture collections ⢠CYP broth was also efficient in reducing environmental contamination in field-capture samples, improving Y. pestis isolation ⢠CYP broth can also be used for the isolation of Y. enterocolitica and Y. pseudotuberculosis.
Assuntos
Peste , Yersinia pestis , Humanos , Ágar , Peste/microbiologia , Novobiocina/farmacologia , Nistatina , Meios de Cultura/farmacologia , Cefsulodina/farmacologiaRESUMO
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)
Assuntos
Humanos , Feminino , Adulto , Nistatina , Vaginose Bacteriana/etiologia , Terapia Combinada , Metronidazol , Nifuratel , Lactobacillus delbrueckiiRESUMO
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.
Assuntos
Antifúngicos , Estomatite sob Prótese , Antifúngicos/farmacologia , Nistatina/farmacologia , Candida , Estomatite sob Prótese/tratamento farmacológico , Estomatite sob Prótese/microbiologia , Testes de Sensibilidade Microbiana , Candida albicans , Candida parapsilosisRESUMO
OBJECTIVES: This randomized clinical trial evaluated the effectiveness of an interim denture resilient liner (Trusoft) modified with minimum inhibitory concentrations (MICs) of antimicrobial agents for Candida albicans biofilm in the denture stomatitis (DS) treatment. METHODS: Forty participants with DS and maxillary complete denture (MCD) wearers were randomly assigned to one of the treatments for 14 days (n=10): nystatin oral suspension (Control-100,000IU/mL; 4 × /day), MCD relined with Trusoft either without (Tru) or with nystatin (Ny) or chlorhexidine diacetate (Chx) at MICs. Cytological smears and mycological quantitative cultures were taken from the palate and denture before treatment (baseline), at the end of treatment (day 14), and at follow-up (days 30, 45, and 60). Photographs of the palate were made at each visit. Data were analyzed by the Cochran and χ2 tests, ANOVA and the Tukey test or the Friedman and Kruskal-Wallis tests (α=0.05). RESULTS: Palatal smears of the Ny and Chx groups exhibited no mycelial Candida (0%) on day 14, and, at the 60-day follow-up, it was observed for only 1 participant from Chx group. MCD smears showed reduction in mycelial forms for all groups on day 14 (P<0.05), but this difference was maintained at follow-up only for the relined dentures (P<0.05). Unlike Tru and the control groups, Ny and Chx groups evidenced a significant reduction in CFU/mL values and DS severity throughout the trial (P<0.05). CONCLUSIONS: The addition of nystatin and chlorhexidine at MICs to an interim resilient liner is a promising treatment for DS, with better results than those for conventional therapy with nystatin suspension, including the follow-ups. CLINICAL SIGNIFICANCE: Compared with conventional topical antifungal therapy, modifying a denture reline material with antimicrobials provided a therapeutic option for denture stomatitis. This non-invasive, straightforward therapeutic approach can be easily adopted by dentists and has the advantage of not requiring patient compliance while maintaining therapeutic concentrations on the denture base.
Assuntos
Nistatina , Estomatite sob Prótese , Humanos , Nistatina/uso terapêutico , Nistatina/farmacologia , Antifúngicos , Estomatite sob Prótese/tratamento farmacológico , Estomatite sob Prótese/microbiologia , Clorexidina/uso terapêutico , Candida albicansRESUMO
This study was performed to evaluate the biocompatibility and antifungal effect of Rosmarinus officinalis against Candida albicans in Galleria mellonella model. Five different concentrations of R. officinalis glycolic extract (50; 25; 12.5 e 6.25 mg/mL) were used to evaluate its biocompatibility in G. mellonella model, in which the nystatin suspension (100; 50; 25; 12.5 e 6.25%) was used as a control group. The antifungal action of R. officinalis glycolic extract was evaluated on C. albicans for 72, 48 and 12 h at two different phases: (1) using the extract as therapeutic agent; and (2) using the extract as prophylactic agent. PBS was used as a negative control group. G. mellonella survival curves were plotted using the Kaplan-Meier method and statistical analysis was performed using the log-rank test (Mantel-Cox) and the significance level was set at (α ≤ 0.05). There was no significant difference among the groups in which all were biocompatible except of a significant death rate of 26.6% with nystatin 100%. In phase 1, it was found that after 7 days, there was no statistically significant difference among the prophylactic treatment groups. In phase 2, the groups of R. officinalis 6.25 mg/mL for 72 h and R. officinalis of 12.5 mg/mL for 24 h promoted the survival rate of the larvae in comparison with the control group with a significant difference (p = 0.017) and (p = 0.032) respectively. Therefore, R. officinalis extract is biocompatible in different concentrations and can be used as a prophylactic agent against fungal infection.
Assuntos
Mariposas , Rosmarinus , Animais , Antifúngicos/farmacologia , Candida albicans , Nistatina/farmacologia , Extratos Vegetais/farmacologiaRESUMO
Amphotericin B is increasingly used in treatment of leishmaniasis. Here, fourteen independent lines of Leishmania mexicana and one L. infantum line were selected for resistance to either amphotericin B or the related polyene antimicrobial, nystatin. Sterol profiling revealed that, in each resistant line, the predominant wild-type sterol, ergosta-5,7,24-trienol, was replaced by other sterol intermediates. Broadly, two different profiles emerged among the resistant lines. Whole genome sequencing then showed that these distinct profiles were due either to mutations in the sterol methyl transferase (C24SMT) gene locus or the sterol C5 desaturase (C5DS) gene. In three lines an additional deletion of the miltefosine transporter gene was found. Differences in sensitivity to amphotericin B were apparent, depending on whether cells were grown in HOMEM, supplemented with foetal bovine serum, or a serum free defined medium (DM). Metabolomic analysis after exposure to AmB showed that a large increase in glucose flux via the pentose phosphate pathway preceded cell death in cells sustained in HOMEM but not DM, indicating the oxidative stress was more significantly induced under HOMEM conditions. Several of the lines were tested for their ability to infect macrophages and replicate as amastigote forms, alongside their ability to establish infections in mice. While several AmB resistant lines showed reduced virulence, at least two lines displayed heightened virulence in mice whilst retaining their resistance phenotype, emphasising the risks of resistance emerging to this critical drug.
Assuntos
Antiprotozoários , Leishmania mexicana , Camundongos , Animais , Anfotericina B/farmacologia , Leishmania mexicana/metabolismo , Nistatina , Soroalbumina Bovina/metabolismo , Esteróis , Estresse Oxidativo , Polienos , Transferases/metabolismo , Glucose , Ácidos Graxos Dessaturases/metabolismo , Antiprotozoários/farmacologiaRESUMO
The objective of this study was evaluate, in vivo model, the antifungal activity of Cryptocarya moschata extract against Candida albicans and its biocompatibility. The animals (N = 50) were divided into groups (n = 5): CI/CG: candidiasis was induced and treated with C. moschata extract (0.045 g/mL); CI/NG: candidiasis was induced and treated with nystatin; CI/NT: candidiasis was induced and no treated; CI/CG-2: candidiasis was induced and treated with C. moschata extract (0.045 g/mL), reapplied after 24 h; CI/NG-2: candidiasis was induced and treated with nystatin, reapplied after 24 h; NCI/NT: candidiasis was not induced and no treated; NCI/CG: candidiasis was not induced and treated with C. moschata extract (0.045 g/mL); NCI/NG: candidiasis was not induced treated with nystatin; NCI/CG-2: candidiasis was not induced and treated with C. moschata extract (0.045 g/mL), reapplied after 24 h; NCI/NG-2: candidiasis was not induced and treated with nystatin, reapplied after 24 h. The fungi present in the lingual dorsum of mice were collected and analyzed by the count of colony-forming units. In addition, histological analysis was performed. Histologically, there was no cell damage in the mice's tongue, and there was a decrease in Candida biofilm, similar to the use of nystatin. It was concluded that the C. moschata extract was effective against C. albicans and was biocompatible.
Assuntos
Antifúngicos , Cryptocarya , Animais , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candida albicans , Camundongos , Testes de Sensibilidade Microbiana , Nistatina/farmacologia , Extratos Vegetais/farmacologiaRESUMO
OBJECTIVE: This study assessed the effects of chitosan (CS) on microcosm biofilms derived from saliva of patients with Candida-associated denture stomatitis. METHODS: Five removable denture wearers with denture stomatitis were included in the study. The minimum inhibitory concentration (MIC) of CS against clinical isolates of Candida albicans was determined according to the broth microdilution method. Pooled saliva from the donors was used as an inoculum for the formation of biofilms, which were developed during 72 h on acrylic surfaces in the Amsterdam Active Attachment model. The biofilms were then treated with different concentrations of CS, and the antibiofilm effects were evaluated through the quantification of colony-forming units (CFUs), total biomass (TB), metabolic activity (MA), lactic acid production (LAP), and cell viability (by confocal laser scanning microscopy). Chlorhexidine, miconazole, and nystatin were tested as positive controls, while the negative control (NC) was the untreated biofilm. Data were analyzed by 1-way ANOVA and Fischer LSD's post hoc test (α=0.05). RESULTS: MIC values of CS ranged from 500 to 800 µg/mL. For CFUs, 2500 µg/mL CS was the most effective treatment in reducing total anaerobes, mutans streptococci, and Lactobacillus spp., significantly differing from the controls. For C. albicans CFUs, CS and positive controls did not differ from each other but led to significant reductions compared to NC. Regarding TB, MA, LAP, and cell viability, 2500 µg/mL CS promoted the greatest reductions compared to NC. CONCLUSION: CS has similar or superior effects to conventional active principles on important parameters of oral candidiasis microcosm biofilms. CLINICAL RELEVANCE: The antibiofilm effects of CS show that this compound has great potential to improve the clinical condition of denture stomatitis patients, and formulations containing this natural polymer could be useful for controlling oral candidiasis.
Assuntos
Candidíase Bucal , Quitosana , Estomatite sob Prótese , Humanos , Resinas Acrílicas/farmacologia , Antifúngicos/farmacologia , Biofilmes , Candida albicans , Candidíase Bucal/tratamento farmacológico , Quitosana/farmacologia , Clorexidina/farmacologia , Ácido Láctico/farmacologia , Miconazol/farmacologia , Nistatina/farmacologia , Estomatite sob Prótese/tratamento farmacológicoRESUMO
Aim: This study investigated the effect of denture liners surface modification with Equisetum giganteum (EG) and Punica granatum (PG) on Candida albicans biofilm inhibition supposing its usage as a sustained-release therapeutical delivery system for Candida-associated denture stomatitis. Materials & methods:C. albicans biofilm (SC5314 or ATCC 90028) was formed on soft liners superficially modified by a primer mixed to drugs at minimum inhibitory concentrations (0.100 g for EG and PG or 0.016 g for nystatin per ml of primer). After 24 h, 7 or 14 days, antibiofilm activity was evaluated by colony-forming unit counts. Results: Not all groups were equi-efficient to nystatin after 24 h and 7 days. After 14 days, EG and PG efficacies were not different from nystatin (almost 100% inhibition). Conclusion: The proposed protocol presents a promising option to allopathic drugs for Candida-associated denture stomatitis treatment.