Análise in vitro da atividade antimicrobiana, citotoxicidade e genotoxicidade de um princípio ativo (Timol) e de um enxaguatório bucal (LISTERINE® ZERO™)

Pós-graduação em Biopatologia Bucal - ICT

Propriedades antimicrobianas, físico-mecânicas e de liberação de fluoreto do cimento de ionômero de vidro associado ao hexametafosfato de sódio microparticulado e nanoparticulado

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito antimicrobiano e citotóxico do óleo essencial de Cymbopogon citratus sobre células odontoblastóides

The aim of this study was to evaluate the antimicrobial and cytotoxic effect of essential oil (EO) of lemon grass (Cymbopogon citratus). From the agar diffusion method, different concentrations of EO (0.135%, 0.2% and 1%), and control solutions (chlorhexidine (Chx), distilled water (Ad) and cereal alcohol (Ac)) were applied on cultures of Candida albicans (C.a), Streptococcus mutans (S.m), Streptococcus sobrinus (S.sob) and Lactobacillus acidophilus (L.a). For C.a, S.m and S.sob, the largest inhibition zones in descending order were: Chx, Ac and EO 1%, while the latter two were statistically similar (Mann-Whitney, p> 0.05). For L.a, the largest inhibition halo was observed for the Chx, followed by EO at 1%, 0.2%, 0.135% and Ac. For evaluation of cytotoxicity, the following groups were set: G1: 0,1% EO; G2: pure EO; G3 (positive control): H2 O2 ; G4: cereal alcohol; and G5 (negative control): culture medium – DMEM. The solutions were applied on the cultured MDPC-23 cells, which were plated (30,000 cells/cm2 ) in wells of 24 well-dishes. Cell metabolism was evaluated by MTT assay. Considering G5 (negative control) as 100% of cell metabolism, it was observed for G1, G2, G3 and G4 a percentage reduction in cell metabolism of 29.6%, 82%, 81.2% and 33.4%, respectively. It was concluded that the low concentration of 0,1% OE (C. citratus) was able to inhibit the growth of the strains tested as well as caused mild cytotoxicity to the cultured MDPC-23 cells.

Incorporation of chlorhexidine gluconate or diacetate into a glass-ionomer cement: porosity, surface roughness, and anti-biofilm activity

To evaluate the porosity, surface roughness and anti-biofilm activity of a glass-ionomer cement (GIC) after incorporation of different concentrations of chlorhexidine (CHX) gluconate or diacetate. Methods: For the porosity and surface roughness tests, 10 test specimens were fabricated of the GIC Ketac Molar Easy Mix (KM) and divided into the following groups: Control, GIC and 0.5% CHX diacetate; GIC and 1.0% CHX diacetate; GIC and 2.0% CHX diacetate; GIC and 0.5% CHX gluconate; GIC and 1.0% CHX gluconate; GIC and 2.0% CHX gluconate. To evaluate porosity, the test specimens were fractured. The fragments were photographed by scanning electron microscopy (SEM), and the images analyzed with the aid of the software program Image J. The surface roughness (Ra) was obtained by the mean value of three readouts performed on the surface of each specimen, always through the center. To analyze the anti-biofilm activity, strains of S. mutans ATCC 35688 were used, and the groups control and GIC +CHX diacetate 1% were divided as follows: GIC (1 day); GIC (7 days), GIC (14 days), GIC (21 days); GIC+CHX (1 day), GIC+CHX (7 days), GIC+CHX (14 days), GIC+CHX (21 days); GIC+ CHX (1 day), GIC+ CHX (7 days), GIC+ CHX (14 days) and GIC+ CHX (21 days) using 10 test specimens per group. For biofilm growth, the specimens were placed in a vertical position in 24-well plates and incubated overnight 10 times. The culture medium was renewed every 24 hours. The suspension was diluted and seeded on BHI agar for quantification of the bacteria present. For evaluation of all the tests the two-way ANOVA was used, and if necessary, the Tukey test was applied, with a level of significance of 5%. Results: Regarding GIC porosity, the ANOVA showed that the presence of CHX increased the porosity (P< 0.001) proportionally to the increase in concentrations (P= 0.001), without however, presenting interaction between material and concentration (P= 0.705). Regarding the number of pores, a significant increase in pores was observed with the increase in CHX concentration (P= 0.003). The surface roughness test demonstrated no statistically significant effect as to increase or reduction in roughness at any of the CHX concentrations used (P> 0.05). Anti-biofilm activity analysis pointed out a significant effect of the factors material (P= 0.006) and time (P< 0.001), with CHX diacetate CHX presenting greater effectiveness in reducing microorganisms.

Effect of light-curing on the antibacterial activity of dentin bonding agents

Aim: This study evaluated the effect of light-activation on the antibacterial activity of dentin bonding systems. Methods: Inocula of Streptococcus mutans and Lactobacillus casei cultures were spread on the surface of BHI agar and the materials were applied and subjected or not to light-activation. Zones of bacterial growth inhibition around the discs were measured. Results: Excite, Single Bond and the bond of Clearfil SE Bond (SE) and Clearfil Protect Bond (CP) did not show any antibacterial activity. The strongest inhibitory activity was observed for the primers of CP and Prompt (PR) against S. mutans and the primers of SE and PB against L. casei. Conclusion: Light-activation significantly reduced or suppressed the antibacterial activity of the initially active uncured dentin bonding systems

Ph and antimicrobial activity of portland cement associated with different radiopacifying agents

The aim of this study was to evaluate the antimicrobial activity and pH changes induced by Portland cement (PC) alone and in association with radiopacifiers. Methods. The materials tested were pure PC, PC + bismuth oxide, PC + zirconium oxide, PC + calcium tungstate, and zinc oxide and eugenol cement (ZOE). Antimicrobial activity was evaluated by agar diffusion test using the following strains: Micrococcus luteus, Streptococcus mutans, Enterococcus faecalis, Pseudomonas aeruginosa, and Candida albicans. After 24 hours of incubation at 37°C, inhibition of bacterial growth was observed and measured. For pH analysis, material samples (n=10) were placed in polyethylene tubes and immersed in 10 mL of distilled water. After 12, 24, 48, and 72 hours, the pH of the solutions was determined using a pH meter. Results. All microbial species were inhibited by the cements evaluated. All materials composed of PC with radiopacifying agents promoted pH increase similar to pure Portland cement. ZOE had the lowest pH values throughout all experimental periods. Conclusions. All Portland cement-based materials with the addition of different radiopacifiers (bismuth oxide, calcium tungstate, and zirconium oxide) presented antimicrobial activity and pH similar to pure Portland cement.

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

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effects of probiotic fermented milk on biofilms, oral microbiota, and enamel

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ação da histatina 5, uma proteína antimicrobiana, no desenvolvimento de biofilmes de Candida Albicans

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Indicadores de qualidade e composição de leite informal comercializado na região Sudeste do Estado de São Paulo

The informal milk is recognized as the product marketed without inspection. We evaluated the microbiological quality, nutritional ingredients and substances inhibiting bacterial growth in 100 informal samples from cows milk, marketed in the southwest region of the state of Sao Paulo, Brazil. It was found that 77% of Somatic Cell Count (SCC) and 86% of Total Bacterial Count (TBC) of the samples were at odds with the maximum values required by Instruction Rules 62 of the Ministry of Agriculture and Livestock Supply. It was identified to solids contents in disagreement (36%), protein (23%), fat (38%), nonfat dry extract (43%) and 73% urea nitrogen. In 59% of the samples was detected residues of the bacterial growth inhibitors. Were isolated 240 strains of micro-organisms with a prevalence of enterobacteria, streptococci, staphylococci and fungi. The informal milk is marketed in the region without the minimum hygienic conditions and present risks to public health, and represents serious socio-economic problem for the region.

Different bacterial models for in vitro induction of non-cavitated enamel caries-like lesions: microhardness and polarized light miscroscopy analyses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Anthocyanin incorporated dental copolymer: bacterial growth inhibition, mechanical properties, and compound release rates and stability by (1)h NMR

Objective. To evaluate bacterial growth inhibition, mechanical properties, and compound release rate and stability of copolymers incorporated with anthocyanin (ACY; Vaccinium macrocarpon). Methods. Resin samples were prepared (Bis-GMA/TEGDMA at 70/30 mol%) and incorporated with 2 w/w% of either ACY or chlorhexidine (CHX), except for the control group. Samples were individually immersed in a bacterial culture (Streptococcus mutans) for 24 h. Cell viability (n = 3) was assessed by counting the number of colony forming units on replica agar plates. Flexural strength (FS) and elastic modulus (E) were tested on a universal testing machine (n = 8). Compound release and chemical stability were evaluated by UV spectrophotometry and (1)H NMR (n = 3). Data were analyzed by one-way ANOVA and Tukey's test ( α = 0.05). Results. Both compounds inhibited S. mutans growth, with CHX being most effective (P < 0.05). Control resin had the lowest FS and E values, followed by ACY and CHX, with statistical difference between control and CHX groups for both mechanical properties (P < 0.05). The 24 h compound release rates were ACY: 1.33 μg/mL and CHX: 1.92 μg/mL. (1)H NMR spectra suggests that both compounds remained stable after being released in water. Conclusion. The present findings indicate that anthocyanins might be used as a natural antibacterial agent in resin based materials.

Antimicrobial activity of a tissue conditioner combined with a biocide polymer

The characteristics of tissue conditioners support microorganism development that can threaten the health of the dentures user. The object of this study was to evaluate the effect on antimicrobial activity, roughness and wettability surface of a tissue conditioners material combined with the antimicrobial polymer poly (2-tert-butilaminoethyl) methacrylate (PTBAEMA). Specimens of tissue conditioner (Coe Soft(®)) were divided into three groups, according to the concentration of PTBAEMA incorporated (0, 10 and 25%). Antimicrobial activity was assessed by adherence assay of one of the microorganisms, Staphylococcus aureus, Streptococcus mutans and Candida albicans. Roughness measurements were made using a Mitutoyo SJ-400, and the mean arithmetic roughness values (Ra) obtained were used for the comparisons. The wettability properties were determined by contact angle measurements. The group containing 25% of PTBAEMA inhibited totally the S. aureus and S. mutans biofilm formation. A significant reduc tion in the S. aureus (Kruskal-Wallis, p = 0,001) and S. mutans (Kruscal-Wallis, p = 0,001) count for 10% PTBAEMA group compared with respective control group. No significant difference was found for C. albicans among PTBAEMA groups and control group (ANOVA, p > 0,05). Incorporating 10 and 25% PTBAEMA increased surface roughness and decreased contact angles (ANOVA and Tukey's post hoc tests, α = 5%). Incorporating 10% PTBAEMA into tissue conditioner increases wettability and roughness of tissue conditioner surface; and decreases the adhesion of S. mutans and S. aureus on material surface, but did not exhibit antimicrobial effect against C. albicans. The PTBAEMA incorporated into tissue conditioner could prevent biofilm formation on elderly patient.

Photodynamic inactivation of a multispecies biofilm using Photodithazine(®) and LED light after one and three successive applications

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Protective efficacy of psidium cattleianum (Sabine) and myracrodruon urundeuva (Allemao) leaf extracts against caries development in rats

This study evaluated the influence of Psidium cattleianum Sabine (Myrtaceae) and Myracrodruon urundeuva Allemão (Anacardiaceae) aqueous extracts on S. mutans counts and dental enamel micro-hardness of rats submitted to a cariogenic challenge. Sixty Wistar rats were distributed in three groups and received water (control) or aqueous extracts of Psidium cattleianum or Myracrodruon urundeuva as hydration solution. Initially the animals had their sublingual and submandibular salivary glands surgically removed and the parotid ducts ligated. Then the rats were inoculated with 106 CFU of Streptococcus mutans ATCC 35668 and were fed with a cariogenic diet. To detect and quantify the presence of S. mutans, oral biofilms were sampled and microbial DNA was extracted and submitted to amplification by means of real-time PCR (Polymerase Chain Reaction). After seven weeks the animals were sacrificed and enamel demineralization was analyzed by cross-sectional micro-hardness. Both extracts produced a significant reduction on S. mutans counts and decreased the enamel demineralization. It can be concluded that the extracts tested had a significant effect on S. mutans in oral biofilm of the rats, decreasing S. mutans accumulation and enamel demineralization.