Antifungal activity of synthetic antiseptics and natural compounds against Candida dubliniensis before and after in vitro fluconazole exposure

ABSTRACT INTRODUCTION: This study evaluated the susceptibilities of oral candidiasis-derived Candida albicans, fluconazole-resistant (FR) Candida dubliniensis, and fluconazole-susceptible (FS) C. dubliniensis to synthetic antiseptics [chlorhexidine gluconate (CHX), cetylpyridinium chloride (CPC), and triclosan (TRC)] and natural compounds (carvacrol, eugenol and thymol). METHODS: Susceptibility tests were performed based on the M27-A3 reference method. The fluconazole-resistant C. dubliniensis strains were obtained after prolonged in vitro exposure to increasing fluconazole concentrations. The geometric mean values for minimum inhibitory concentrations and minimum fungicidal concentrations were compared among the groups. RESULTS: Fluconazole-susceptible C. dubliniensis was more sensitive to CPC and TRC than FR C. dubliniensis and C. albicans were. However, eugenol and thymol were more active against FR C. dubliniensis. The fungicidal activities of CHX and TRC were similar for the three groups, and FR C. dubliniensis and C. albicans had similar sensitivities to CPC. CONCLUSIONS: The resistance of C. dubliniensis to fluconazole affects its sensitivity the synthetic antiseptics and natural compounds that were tested.

Effect of antibacterial agents on the surface hardness of a conventional glass-ionomer cement

In atraumatic restorative treatment (ART), caries removal with hand excavation instruments is not as efficient as that with rotary burs in eliminating bacteria under the glass ionomer cements (GICs). Thus, different antibacterial agents have been used in recent studies to enhance the antibacterial properties of the GICs, without jeopardizing their basic physical properties. OBJECTIVE: The objective of this study was to evaluate the effect of antibacterial agents on the surface hardness of a conventional GIC (Fuji IX) using Vickers microhardness [Vickers hardness number (VHN)] test. MATERIAL AND METHODS: Cetrimide (CT), cetylpyridinium chloride (CPC) and chlorhexidine (CHX) were added to the powder and benzalkonium chloride (BC) was added to the liquid of Fuji IX in concentrations of 1 percent and 2 percent, and served as the experimental groups. A control group containing no additive was also prepared. After the completion of setting reaction, VHN measurements were recorded at 1, 7, 15, 30, 60, and 90 days after storage in 37°C distilled water. A one-way ANOVA was performed followed by a Dunnett t test and Tamhane T2 tests and also repeated measurements ANOVA was used for multiple comparisons in 95 percent confidence interval. RESULTS: VHN results showed significant differences between the control and the experimental groups at all time periods (p<0.05 for all). Significant differences were observed between all study periods for individual groups (p<0.05). After 7 days, VHNs were decreased in all experimental groups while they continued to increase in the control group. BC and CHX groups demonstrated the least whereas CT and CPC groups exhibited most adverse effect on the hardness of set cements. CONCLUSIONS: Despite the decreased microhardness values in all experimental groups compared to the controls after 7 up to 90 days, incorporating certain antibacterial agents into Fuji IX GIC showed tolerable microhardness alterations within the limitations of this in vitro study.

A preliminary study of the antibacterial potential of cetylpyridinium chloride in root canals infected by E. faecalis

The aim of this preliminary study was to verify the antibacterial potential of cetylpyridinium chloride (CPC) in root canals infected by Enterococcus faecalis. Forty human maxillary anterior teeth were prepared and inoculated with E. faecalis for 60 days. The teeth were randomly assigned to the following groups: 1: Root canal preparation (RCP) + 0.1% CPC with positive-pressure irrigation (PPI, Conventional, NaviTip®); 2: RCP + 0.2% CPC PPI; 3: RCP + 2.5% NaOCl PPI; 4: RCP + 2.5% NaOCl with negative-pressure irrigation system (NPI, EndoVac®); 5: Positive control; and 6: Negative control. Four teeth of each experimental group were evaluated by culture and 4 by scanning electron microscopy (SEM). In all teeth, the root canals were dried and filled with 17% EDTA (pH 7.2) for 3 min for smear layer removal. Samples from the infected root canals were collected and immersed in 7 mL of Letheen Broth (LB), followed by incubation at 37°C for 48 h. Bacterial growth was analyzed by turbidity of culture medium and then observed with a UV spectrophotometer. The irrigating solutions were further evaluated for antimicrobial effect by an agar diffusion test.The statistical data were treated by means, standard deviation, Kruskal-Wallis test and analysis of variance. Significance level was set at 5%. The results showed the presence of E. faecalis after root canal sanitization. The number of bacteria decreased after the use of CPC. In the agar diffusion test, CPC induced large microbial inhibition zones, similar to 2% chlorhexidine and large than 2.5% NaOCl. In conclusion, cetylpyridinium chloride showed antibacterial potential in endodontic infection with E. faecalis.

O objetivo deste estudo preliminar foi verificar o potencial antibacteriano de cloreto de cetilpiridínio (CCP) em canais radiculares infectados por E. faecalis. Quarenta dentes anteriores de humanos foram preparados e inoculados com E. faecalis por 60 dias. Os dentes foram aleatoriamente distribuídos como se segue: 1. Preparo do canal radicular (PCR) + CCP 0,1% com sistema de pressão positiva de irrigação (PPI, convencional, Navitip®); 2. PCR + CPC 0,2% PPI; 3. PCR + NaOCl 2,5% PPI, 4. PCR + NaOCl 2,5% com sistema de pressão negativa de irrigação (PNI, EndoVac®); 5 e 6. Controles positivos e negativos. Quatro dentes de cada grupo experimental foram avaliados por cultura e quatro por microscopia eletrônica de varredura (MEV). Em todos os dentes, os canais foram secos e preenchidos com EDTA 17% (pH 7,2) durante 3 min. As amostras dos canais radiculares infectados foram coletadas e imersas em 7 mL Letheen Broth (LB), seguido de incubação a 37° C durante 48 h. O crescimento bacteriano foi analisado pela turvação do meio de cultura, e mensurados por meio de um espectrofotometro (UV). As soluções irrigantes foram ainda avaliadas em teste de difusão em ágar. A análise estatística utilizou de média, desvio padrão,teste de Kruskal-Wallis e análise de variância. O nível de significância foi de 5%. Os resultados mostraram a presença de E. faecalis posterior ao processo de desinfecção do canal radicular. O cloreto de cetilpiridínio mostrou reduzir o número de bactérias. No teste de difusão em ágar, o CPC determinou inibição microbiana, com resultados semelhantes à CHX a 2% e maiores do que o hipoclorito de sódio a 2,5%. O cloreto de cetilpiridínio demonstrou potencial antibacteriano em infecção endodôntica por E. faecalis.

Prevalence of mutans streptococci isolated from complete dentures and their susceptibility to mouthrinses

The aims of this study were to evaluate the incidence of mutans streptococci (MS - sessile form) on complete maxillary dentures after use of a specific denture paste, and to determine the minimum inhibitory concentration (MIC) and maximum inhibitory dilution (MID) of 3 oral mouthrinses: Cepacol, Plax and Periogard. Seventy-seven complete denture wearers were randomly assigned into 2 groups, according to the product used for denture cleaning: Control group - conventional dentifrice (Kolynos-Super White); and Test group: experimental denture cleaning paste. Denture biofilm was collected at baseline and after 90 and 180 days after treatment by brushing the dentures with saline solution. After decimal serial dilution, samples were seeded onto agar sucrose bacitracin to count colonies with morphological characteristics of MS. MS identification was performed by the sugar fermentation tests. After this procedure, brain heart infusion broth (BHI) was added to oral mouthrinses (Plax, Cepacol e Periogard) and seeded on Petri dishes. The colonies were seeded using the Steers multiplier and, after the incubation, the MIC and MID of the mouthrinses were calculated. The results showed an incidence of 74.0 percent (n=57) of MS in the 77 complete dentures examined in the study, being 76.3 percent (n=29) of the Control group (conventional dentifrice) and 71.8 percent (28) of the Test group (experimental denture cleaning paste). In both groups, the number of positive cases for MS decreased from day 0 to day 180. In the Test group there was a slight decrease in the incidence of Streptococcus mutans 90 days after use of the experimental denture cleaning paste, which was not observed in the Control group. As regards to mouthrinses, for both groups, Periogard showed antimicrobial action with the highest dilution, followed by Cepacol and Plax. In conclusion, the incidence of MS in complete dentures was high and Periogard was the mouthrinse with the strongest antimicrobial action against MS. The experimental denture cleaning paste showed a slight action against S. mutans after 90 days of treatment.

O objetivo deste estudo foi avaliar a incidência de estreptococos do grupo mutans (forma séssil) de próteses totais superiores após o uso de uma pasta específica para higienização de próteses totais, bem como determinar a concentração inibitória mínima (CIM) e diluição inibitória máxima (DIM) de 3 enxaguatórios bucais: Cepacol, Plax e Periogard. Setenta e sete usuários de próteses totais foram distribuídos em 2 grupos: (A) grupo Controle: dentifrício convencional (Kolynos Super Branco) e (B) grupo Teste: pasta experimental para higiene de próteses (1). O biofilme das próteses foi coletado no início e após 90 e 180 dias dos tratamentos, por meio de escovação das próteses com solução salina. Após a diluição decimal seriada, as amostras foram semeadas em agar sacarose bacitracina para a contagem de colônias características de estreptococos do grupo mutans. A identificação dos estreptococos do grupo mutans foi realizada por meio de testes de fermentação de açúcares. Após este procedimento, o caldo de infusão de cérebro e coração (BHI) foi adicionado aos antissépticos (Plax, Cepacol e Periogard) e semeado em placas de Petri. As colônias foram semeadas usando o mutiplicador de Steers e após a incubação, a concentração inibitória mínima e a concentração inibitória máxima dos enxaguatórios foram calculadas. Os resultados mostraram que a incidência de estreptococos do grupo mutans nas 77 próteses totais examinadas foi de 74,0 por cento (n=57), sendo 76,3 por cento (n=29) do grupo Controle (pasta convencional) e 71,8 por cento (n=28) do grupo Teste (pasta experimental). Em ambos os grupos, o número de casos positivos para estreptococos do grupo mutans diminuiu, do início (0 dias) para 180 dias. Houve uma pequena diminuição da incidência de Streptococcus mutans após 90 dias do uso da pasta experimental, o que não foi observado no grupo controle. Com relação aos enxaguatórios, para ambos os grupos, o Periogard apresentou atividade antimicrobiana com diluições maiores, seguido do Cepacol e Plax. Pôde-se concluir que a incidência de estreptococos do grupo mutans em próteses totais foi elevada e que o Periogard foi o enxaguatório com melhor ação antimicrobiana contra os estreptococos do grupo mutans. A pasta experimental mostrou uma pequena ação contra S. mutans após 90 dias de tratamento.

Transport and storage of sputum specimen by using cetylpyridinium chloride for isolation of mycobacteria.

Of the 191 sputum specimens that were collected from pulmonary tuberculosis patients, 78.65% (140/178) specimens were culture positive when processed within 48 h by the NaOH method. The culture positivity in the same specimen that were stored with cetylpyridinium chloride (CPC) and processed after 7-8 days was 70.22% (125/178), whereas those stored without CPC and processed by the NaOH method was 46.62% (83/178). The difference in number of positive cultures obtained before storage and after storage (without CPC) was statistically significant (P = 0.001). Culture positivity by the CPC method was comparable with that of NaOH method before storage and the difference was not statistically significant (P = 0.35).

Determination of the maximum inhibitory dilution of cetylpyridinium chloride-based mouthwashes against staphylococcus aureus: an in vitro study

The aim of this in vitro study was to determine the maximum inhibitory dilution (MID) of four cetylpyridinium chloride (CPC)-based mouthwashes: CPC+Propolis, CPC+Malva, CPC+Eucaliptol+Juá+Romã+Propolis (Natural Honey®) and CPC (Cepacol®), against 28 Staphylococcus aureus field strains, using the agar dilution method. Decimal dilutions ranging from 1/10 to 1/655,360 were prepared and added to Mueller Hinton Agar. Strains were inoculated using Steers multipoint inoculator. The inocula were seeded onto the surface of the culture medium in Petri dishes containing different dilutions of the mouthwashes. The dishes were incubated at 37ºC for 24 h. For readings, the MID was considered as the maximum dilution of mouthwash still capable of inhibiting microbial growth. The obtained data showed that CPC+Propolis had antimicrobial activity against 27 strains at 1/320 dilution and against all 28 strains at 1/160 dilution, CPC+Malva inhibited the growth of all 28 strains at 1/320 dilution, CPC+Eucaliptol+Juá+Romã+Propolis inhibited the growth of 2 strains at 1/640 dilution and all 28 strains at 1/320 dilution, and Cepacol® showed antimicrobial activity against 3 strains at 1/320 dilution and against all 28 strains at 1/160 dilution. Data were submitted to Kruskal-Wallis test, showing that the MID of Cepacol® was lower than that determined for the other products (p<0.05). In conclusion, CPC-mouthwashes showed antimicrobial activity against S. aureus and the addition of other substances to CPC improved its antimicrobial effect.

Determinaçäo do consumo de glocose "in vitro" pelos microorganismos presentes na saliva total de humanos. I - Açäo do cloreto de cetilpiridínio e do triclosan/gantrez / Determination of the consumptionof glicose \"in vitro\" by microorganisms present in the total salive of human. I - Action of cetyl-pyridinium chloride and triclosan/gantrez

A via glicolítica é um caminho metabólico utilizado pelas células em geral, e particularmente pelos microorganismos, visando a obtençäo de energia. A cárie dentária e a doença periodontal estäo diretamente relacionadas à placa bacteriana. Tanto a placa recente quanto a placa näo perturbada têm na glicose o substrato necessário às atividades metabólicas, cuja patogenicidade é caracterizada pela intensidade de ácidos resultantes da utilizaçäo deste substrato. O íon fluoreto e os antissépticos cloreto de cetilpiridinio e triclosan / gantrez apresentam, isoladamente ou de forma associada, propriedades inibidoras dos microorganismos. O presente estudo realizado "in vitro" demonstra a açäo destes produtos farmacológicos sobre o consumo de glicose pelos microorganismos presentes na saliva total de humanos. Foram testadas soluçöes de fluoreto de sódio - 226ppm. de cloreto de cetilpiridinio monohidratado a 0,05 porcento e das associaçöes de cloreto de cetilpiridinio monohidratado a 0,05 porcento / fluoreto de sódio - 228 ppm e de triclosan a 0,03 porcento, gantrez a 0,15 porcento e fluoreto de sódio - 225ppm. Os resultados experimentais expressam, através do consumo de glicose pelos microorganismos presentes na saliva total, inibiçäo estatisticamente significante, causada por estas substâncias medicamentosas a partir de 180 minutos após o contato entre estes fármacos e a mistura saliva total / glicose. Näo se observou diferentes eficácias significantes entre estes fármacos no processo de inibiçäo do consumo da glicose pelos microorganismos (Kruskal-Wallis, ANOVA seguido pelo Mann Whitney). Esta atividade inibidora, atribuída ao fluoreto de sódio, ao cloreto de cetilpiridinio e às associaçöes de cloreto de cetilpiridinio monohidratado / fluoreto de sódio e de triclosan / gantrez / fluoreto de sódio, contribui sobremaneira na prevençäo da cárie dentária e da doença periodontal