Effect of iodonium salt and chitosan on the physical and antibacterial properties of experimental infiltrants

Abstract Resinous infiltrants are indicated in the treatment of incipient carious lesions, and further development of these materials may contribute to greater control of these lesions. The aim of this study was to analyze the physical and antibacterial properties of experimental infiltrants containing iodonium salt and chitosan. Nine experimental infiltrants were formulated by varying the concentration of the diphenyliodonium salt (DPI) at 0, 0.5 and 1 mol%; and chitosan at 0, 0.12 and 0.25 g%. The infiltrants contained the monomeric base of triethylene glycol dimethacrylate and bisphenol-A dimethacrylate ethoxylate in a 75 and 25% proportion by weight, respectively; 0.5 mol% camphorquinone and 1 mol% ethyl 4-dimethylaminobenzoate. The degree of conversion was evaluated using Fourier transformer infrared spectroscopy, and the flexural strength and elastic modulus using the three-point bending test. Sorption and solubility in water, and antibacterial analysis (minimum inhibitory concentration and minimum bactericidal concentration) were also analyzed. Data was analyzed statistically by two-way ANOVA and Tukey's test (p<0.05), with the exception of the antibacterial test, which was evaluated by visual inspection. In general, the infiltrant group containing 0.5% DPI and 0.12% chitosan showed high values of degree of conversion, higher values of elastic modulus and flexural strength, and lower sorption values in relation to the other groups. Antibacterial activity was observed in all the groups with DPI, regardless of the concentration of chitosan. The addition of DPI and chitosan to experimental infiltrants represents a valid option for producing infiltrants with desirable physical and antibacterial characteristics.

In vitro and in vivo evaluations of glass-ionomer cement containing chlorhexidine for Atraumatic Restorative Treatment

Abstract Objectives: Addition of chlorhexidine has enhanced the antimicrobial effect of glass ionomer cement (GIC) indicated to Atraumatic Restorative Treatment (ART); however, the impact of this mixture on the properties of these materials and on the longevity of restorations must be investigated. The aim of this study was to evaluate the effects of incorporating chlorhexidine (CHX) in the in vitro biological and chemical-mechanical properties of GIC and in vivo clinical/ microbiological follow-up of the ART with GIC containing or not CHX. Material and Methods: For in vitro studies, groups were divided into GIC, GIC with 1.25% CHX, and GIC with 2.5% CHX. Antimicrobial activity of GIC was analyzed using agar diffusion and anti-biofilm assays. Cytotoxic effects, compressive tensile strength, microhardness and fluoride (F) release were also evaluated. A randomized controlled trial was conducted on 36 children that received ART either with GIC or GIC with CHX. Saliva and biofilm were collected for mutans streptococci (MS) counts and the survival rate of restorations was checked after 7 days, 3 months and one year after ART. ANOVA/Tukey or Kruskal-Wallis/ Mann-Whitney tests were performed for in vitro tests and in vivo microbiological analysis. The Kaplan-Meier method and Log rank tests were applied to estimate survival percentages of restorations (p<0.05). Results: Incorporation of 1.25% and 2.5% CHX improved the antimicrobial/anti-biofilm activity of GIC, without affecting F release and mechanical characteristics, but 2.5% CHX was cytotoxic. Survival rate of restorations using GIC with 1.25% CHX was similar to GIC. A significant reduction of MS levels was observed for KM+CHX group in children saliva and biofilm 7 days after treatment. Conclusions: The incorporation of 1.25% CHX increased the in vitro antimicrobial activity, without changing chemical-mechanical properties of GIC and odontoblast-like cell viability. This combination improved the in vivo short-term microbiological effect without affecting clinical performance of ART restorations.

Antimicrobial activity of ozone and NaF-chlorhexidine on early childhood caries

Abstract An early childhood carie (ECC) is an extremely destructive form of tooth decay. The aim of this study was to investigate the action of ozone (O3), and the association of sodium fluoride (NaF) with chlorhexidine (CHX) on bacteria related to ECC. Overnight culture of the bacteria was performed. On exponential phase the suspension was adjusted (101-108 CFU/mL). A drop (10μL) of each concentration of bacteria was applied on sheep blood agar plates and treated with O3 (2, 20, 200, and 2,000 ppm); after 18 hours, recovery analysis of CFU verified the reduction of bacterial activity. For NaF-CHX, sterile 96-well plates were prepared and divided into groups: G1 (150 µL TSB); G2 (20 µL of bacteria + 25 µL CHX + 25 µL NaF); and G3 (150 µL TSB + 20 µL of bacteria + 50 µL water). The plates were verified by analysis of the optical density (0, 12, 14, 16, and 18 hours). The data from O3 test were submitted to ANOVA and Tukey’s test (p < 0.05). For the data from NaF-CHX, the ANOVA 2-way and Bonferroni’s test (p < 0.05) were used. The number of CFU/mL showed death > 3log10 (99.9%) for all bacteria (ozone ≥ 20ppm), while the combination of NaF-CHX was more effective (p < 0.001) compared to each substance tested alone and the control group. The antimicrobial agents tested were able to inhibit all bacteria tested; O3 seemed to be a good alternative for controlling progression of carious lesions, while the association of NaF-CHX showed to be a good antimicrobial with easy and inexpensive application.

Cymbopogon citratus essential oil: effect on polymicrobial caries-related biofilm with low cytotoxicity

Abstract The objective of this study was to evaluate the effects of Cymbopogon citratus essential oil and its main compound (citral) against primary dental colonizers and caries-related species. Chemical characterization of the essential oil was performed by gas chromatography/mass spectroscopy (GC/MS), and the main compound was determined. Antimicrobial activity was tested against Actinomyces naeslundii, Lactobacillus acidophilus, S. gordonii, S. mitis, S. mutans, S. sanguinis and S. sobrinus. Minimum inhibitory and bactericide concentrations were determined by broth microdilution assay for streptococci and lactobacilli reference, and for clinical strains. The effect of the essential oil on bacterial adhesion and biofilm formation/disruption was investigated. Negative (without treatment) and positive controls (chlorhexidine) were used. The effect of citral on preformed biofilm was also tested using the same methodology. Monospecies and microcosm biofilms were tested. ANOVA or Kruskal-Wallis tests were used (α=0.05). Cytotoxicity of the essential oil to human keratinocytes was performed by MTT assay. GC/MS demonstrated one major component (citral). The essential oil showed an inhibitory effect on all tested bacterial species, including S. mutans and L. acidophilus. Essential oil of C. citratus (10X MIC) reduced the number of viable cells of lactobacilli and streptococci biofilms (p < 0.05). The essential oil inhibited adhesion of caries-related polymicrobial biofilm to dental enamel (p < 0.01). Citral significantly reduced the number of viable cells of streptococci biofilm (p < 0.001). The essential oil showed low cytotoxicity to human keratinocytes. Based on these findings, this study can contribute to the development of new formulations for products like mouthwash, against dental biofilms.

Addition of Silver Nanoparticles to Composite Resin: Effect on Physical and Bactericidal Properties In Vitro

The objectives of this study were to evaluate physical properties and antibacterial activity of a light-activated composite modified with silver nanoparticles. Discs were produced with unmodified resin (control group - CG) and modified resin with silver nanoparticles at two concentrations, 0.3% wt (MR03) and 0.6% wt (MR06). Streptococcus mutans and Lactobacillus acidophilus biofilms were induced in vitro by incubation of discs in a 20% sucrose medium, followed by sonication and counting of viable cells after 1, 4 and 7 days (n=9). The arithmetic roughness of all three groups was evaluated by atomic force microscopy (n=9). Compression assay was conducted in all groups to measure the compressive strength at failure and elasticity modulus (n=5). Data were subjected to ANOVA and Tukey's tests (α=0.05%). At all three time points the number of viable cells was statistically lower for MR03 and MR06 compared with CG, for both specimens. MR03 and MR06 showed no significant differences. Microscopic analysis demonstrated no significant differences for roughness among the three groups (p>0.05). The MR03 was stronger to compression than CG, and MR06 was statistically lower than CG and MR03. It was concluded that the MR03 were less conducive to biofilm growth, without compromising the strength in compression and surface roughness.

Os objetivos do estudo foram avaliar as propriedades físicas e a atividade antibacteriana de uma resina composta fotopolimerizável modificada com nanopartículas de prata. Discos foram produzidos com resina não modificada (grupo controle - GC) e resina modificada com nanopartículas de prata em duas concentrações 0,3 e 0,6% em massa (MR03 e MR06, respectivamente). Biofilmes de Streptococcus mutans e Lactobacillus acidophilus foram induzidos in vitro pela incubação dos discos em meio líquido com 20% de sacarose, seguida pela sonicação e contagem de células viáveis após 1, 4 e 7 dias. A rugosidade aritmética de todos os grupos foi avaliada por microscopia de força atômica (MFA). Os dados foram submetidos à ANOVA e ao Teste de Tukey (α=0,05%). Nos três períodos de tempo, o número de células viáveis foi estatisticamente mais baixo para MR03 e MR06, em comparação com GC, para ambas as espécies. MR03 e MR06 não apresentaram diferenças significantes. A análise por meio de MFA demonstrou que não houve diferença significante para a rugosidade entre os três grupos. MR03 apresentouse mais resistente em compressão do que GC, e o desempenho em compressão em MR06 foi estatisticamente pior do que em GC e MR03. Foi possível concluír que as resinas modificadas de MR03 inibiram o crescimento dos biofilmes, sem comprometimento da resistência à compressão e da rugosidade superficial.

Addition of Chlorhexidine Gluconate to a Glass Ionomer Cement: A Study on Mechanical, Physical and Antibacterial Properties

The objective of this work was to determine the effect of different concentrations of chlorhexidine digluconate (CHX) on setting time, surface hardness, maximum tensile bond strength and antibacterial activity of a glass ionomer cement (GIC). The material used as control was Ketac Molar Easymix GIC. CHX was incorporated into the GIC during its manipulation at concentrations of 0.5, 1.0 and 2.0%. Antimicrobial activity against S. mutans and L. acidophilus was evaluated by means of agar diffusion test. Tensile bond strength data were analyzed statistically using Analysis of variance and Tukey's test. Setting time, Vickers hardness and agar diffusion test were analyzed using Kruskal-Wallis and Mann-Whitney tests at a significance level of 5%. It was observed that adding CHX at concentrations of 1% and 2% increased significantly the setting time of the material (p=0.012 and p=0.003, respectively). There was no significant difference between control and 0.5% CHX groups regarding the setting time. Addition of 2% CHX decreased significantly the surface hardness in relation to the control group (p=0.009), followed by the 1% CHX group (p=0.009). The tensile bond strength of the material also decreased significantly after adding CHX at a concentration of 2% (p=0.001). Addition of CHX promoted formation of an inhibition halo in both bacterial strains for all concentrations. The results showed that the best option for clinical use of GIC with CHX is at 0.5% concentration, since antibacterial activity increased and the physical-mechanical properties remained unchanged.

O objetivo deste trabalho foi determinar o efeito de diferentes concentrações de gluconato de clorexidina (CLX) sobre o tempo de presa, dureza superficial, resistência máxima a tração e atividade antibacteriana de um cimento de ionômero de vidro (CIV). O material utilizado como controle foi o CIV Ketac Molar Easymix. O gluconato de CHX foi incorporado no CIV durante a sua manipulação, em concentrações de 0,5, 1,0 e 2,0%. Atividade antimicrobiana contra S. mutans e L. acidophilus foi avaliada por meio de teste de difusão em ágar. Análise de variância (ANOVA) e teste de Tukey foram usadas para avaliar a resistência a tração do material. Para avaliar tempo de presa, dureza Vickers e teste de difusão em ágar foram usados os testes de Mann-Whitney e Kruskal-Wallis ao nível de significância de 5%. Observou-se que a adição de CHX, em concentrações de 1% e 2%, aumentou significativamente o tempo de presa do material (p = 0,012 e p = 0,003, respectivamente). Não houve diferença significativa entre os grupos controle e CHX 0,5% em relação ao tempo de presa. A adição de 2% de CHX diminuiu significativamente a dureza superficial, em relação ao grupo controle (p = 0,009), seguido pelo grupo 1% de CHX (p = 0,009). A resistência à tração do material também diminuiu significativamente após a adição de CHX a uma concentração de 2% (p = 0,001). A adição de CHX promoveu formação de halo de inibição em ambas as linhagens bacterianas para todas as concentrações. Os resultados mostraram que a melhor opção para o uso clínico de CIV com CHX está numa concentração de 0,5%, uma vez que a atividade antibacteriana aumentada e as propriedades físico-mecânicas permaneceram inalteradas.

Chitosan-based polyherbal toothpaste: As novel oral hygiene product.

Objective: The objective of the present work was to develop chitosan-based polyherbal toothpaste and evaluate its plaque-reducing potential and efficacy in reduction of dental pathogens. Materials and Methods: Antimicrobial activity of herbal extracts against dental pathogens were performed by using disk diffusion method. The pharmaceutical evaluation of toothpaste was carried out as per the US Government Tooth Paste Specifications. A 4-week clinical study was conducted in patients with oro-dental problems to evaluate the plaque removing efficacy of chitosan-based polyherbal toothpaste with commercially available chlorhexidine gluconate (0.2% w/v) mouthwash as positive control. Total microbial count was carried out to determine the percentage decrease in the oral bacterial count over the period of treatment. Result: Herbal extracts were found to possess satisfactory antimicrobial activity against most of the dental pathogens. Chitosan-containing polyherbal toothpaste significantly reduces the plaque index by 70.47% and bacterial count by 85.29%, and thus fulfills the majority of esthetic and medicinal requirements of oral hygiene products. Conclusion: Chitosan-based polyherbal toothpaste proves itself as a promising novel oral hygiene product as compared with currently available oral hygiene products. A further study to confirm the exact mechanism and active constituents behind antiplaque and antimicrobial activity of chitosan-based polyherbal toothpaste and its efficacy in large number of patient population is on high demand.

In vitro evaluation of antibacterial activity of an herbal dentifrice against Streptococcus mutans and Lactobacillus acidophilus.

Antibacterial activity of a herbal dentifrice Arodent against Streptococcus mutans and Lactobacillus acidophilus was evaluated using Colgate as standard. Both bacterial strains were isolated from the oral cavity on selective media and identified by standard methods. The antibacterial activity was assayed by cup-well method. The bacterial lawn of facultative anaerobe S. mutans was established between two layers of agar under microaerophilic conditions. Five and a half millimeters and 10 mm zones of inhibition were produced by Arodent against S. mutans and L. acidophilus , respectively, under microaerophilic conditions. On the other hand, the standard dentifrice Colgate produced 5.83 mm and 10.17 mm zones of inhibition against S. mutans and L. acidophilus , respectively, under microaerophilic condition. The results suggest that Arodent is an effective antibacterial herbal dentifrice.

Comparison of antibacterial properties of two fluoride-releasing and a nonfluoride-releasing pit and fissure sealants.

Occlusal pits and fissures are the most susceptible sites for the development of dental caries and prevention is difficult in the areas of occurrence. Although only 12.5% of all the tooth surfaces are occlusal, these surfaces develop more than two-thirds of total caries experienced by children. The fate of the bacteria is of significance when they are inadvertently sealed by a dental sealant. This study aimed in examining the antibacterial properties of three commercially available sealants, viz., Teethmate-F, Helioseal-F and Helioseal. Antibacterial testing was done by disc diffusion method. Selective agars were used to grow Streptococcus mutans and Lactobacillus acidophilus, which are implicated in the pathogenesis of dental caries. Statistical analysis was carried out using student's t test. The study proved that Helioseal-F and Helioseal did not possess antibacterial properties and Teethmate-F was the only sealant that showed antibacterial properties.

Antibiotic sensitivity pattern of various Lactobacillus acidophilus strains.

Seven L. acidophilus strains were examined for their antibiotic sensitivity against various chemotherapeutic agents and all of them were found to be sensitive to novobicin, chloramphenicol, ampicillin, erythromycin, oxytetracycline and chlorotetracycline, whereas, all were resistant to norfloxacin and nalidixic acid. The sensitivity towards remaining antibiotics varied among the strains. The results show that cultures should be tested for their sensitivity towards commonly used chemotherapeutic agents to eliminate starter failure during manufacture of cultured milk products.

Inhibición del crecimiento bacteriano "in vitro" producido por un medio de cementación temporal experimental / In vitro bacterial growth inhibition produced by and experimental temporary luting cement

Se demostró la actividad antibacteriana "in vitro" de un medio de cementación temporal experimental, en base a óxido de zinc eugenol e hidróxido de calcio, frente a una cepa de S. mutans y frente a una de L. acidophillus, ambos microorganismos indicados como algunos de los principales en la producción de la caries dentaria. El medio de cementación experimental presentó inhibición del desarrollo bacteriano "in vitro", tanto frente al Streptococcus mutans como frente al Lactobacillus acidophillus