Chlorhexidine with or without alcohol against biofilm formation: efficacy, adverse events and taste preference

Abstract In recent years, different chlorhexidine formulations have been tested, including an alcohol-free alternative, but the effect of this solution on early biofilm formation is not clear. A crossover, randomized, double-blind clinical trial was conducted to evaluate the effect of two chlorhexidine solutions against supra- and subgingival biofilm formation (NCT#02656251). Thirty-five participants were randomized and asked to rinse twice daily with 15 ml of an alcohol-containing 0.12% chlorhexidine solution, an alcohol-free 0.12% chlorhexidine solution, or placebo. The study was conducted in three experimental periods of 4 days each, with a 10-day washout between the periods. All the experimental periods followed the same protocol, except that the solutions were switched. Biofilm distribution was evaluated every 24 hours by the Plaque-Free Zone Index, during 96 hours. Adverse events were self-reported and sensory evaluation was performed using a hedonic scale. Compared to the placebo, the chlorhexidine solutions resulted in a significantly higher number of surfaces free of plaque over 96 hours (p < 0.01), and were able to prevent subgingival biofilm formation (p < 0.01). The alcohol-free chlorhexidine solution was associated with a lower incidence of adverse events, compared with alcohol-containing chlorhexidine (p < 0.05); it also received better sensory evaluation and acceptance by trial participants, compared with the alcohol-containing chlorhexidine (p = 0.007), and had a similar inhibitory effect on the formation of supra- and subgingival biofilms.

Effect of green tea extract on bonding durability of an etch-and-rinse adhesive system to caries-affected dentin

ABSTRACT Objective Green tea extract has been advocated as a matrix metalloproteinase (MMP) inhibitor; however, its effect on bond durability to caries-affected dentin has never been reported. Thus, the aim of this in vitro study was to evaluate the effect of two MMP inhibitors (2% chlorhexidine and 2% green tea extract), applied after acid etching, on bond durability of an etch-and-rinse adhesive system to caries-affected dentin. Material and Methods Occlusal enamel was removed from third molars to expose the dentin surface, and the molars were submitted to a caries induction protocol for 15 days. After removal of infected dentin, specimens were conditioned with 37% phosphoric acid (15 seconds) and randomly divided into three groups, according to the type of dentin pretreatment (n=10): NT: no treatment; GT: 2% green tea extract; CLX: 2% chlorhexidine. The etch-and-rinse adhesive system (Adper™ Single Bond 2, 3M ESPE, St. Paul, MN, USA) was applied according to the manufacturer's instructions, and composite resin restorations were built on the dentin. After 24 hours, at 37°C, the resin-tooth blocks were sectioned perpendicularly to the adhesive interface in the form of sticks (0.8 mm2 of adhesive area) and randomly subdivided into two groups according to when they were to be submitted to microtensile bond strength (μTBS) testing: immediately or 6 months after storage in distilled water. Data were reported in MPa and submitted to two-way ANOVA for completely randomized blocks, followed by Tukey’s test (α=0.05). Results After 24 hours, there was no significant difference in the μTBS of the groups. After 6 months, the GT group had significantly higher μTBS values. Conclusion It was concluded that the application of 2% green tea extract was able to increase bond durability of the etch-and-rinse system to dentin. Neither the application of chlorhexidine nor non-treatment (NT - control) had any effect on bond strength after water storage.

Antiseptics and microcosm biofilm formation on titanium surfaces

Abstract Oral rehabilitation with osseointegrated implants is a way to restore esthetics and masticatory function in edentulous patients, but bacterial colonization around the implants may lead to mucositis or peri-implantitis and consequent implant loss. Peri-implantitis is the main complication of oral rehabilitation with dental implants and, therefore, it is necessary to take into account the potential effects of antiseptics such as chlorhexidine (CHX), chloramine T (CHT), triclosan (TRI), and essential oils (EO) on bacterial adhesion and on biofilm formation. To assess the action of these substances, we used the microcosm technique, in which the oral environment and periodontal conditions are simulated in vitro on titanium discs with different surface treatments (smooth surface - SS, acid-etched smooth surface - AESS, sand-blasted surface - SBS, and sand-blasted and acid-etched surface - SBAES). Roughness measurements yielded the following results: SS: 0.47 µm, AESS: 0.43 µm, SB: 0.79 µm, and SBAES: 0.72 µm. There was statistical difference only between SBS and AESS. There was no statistical difference among antiseptic treatments. However, EO and CHT showed lower bacterial counts compared with the saline solution treatment (control group). Thus, the current gold standard (CHX) did not outperform CHT and EO, which were efficient in reducing the biofilm biomass compared with saline solution.

How erosive drinks and enzyme inhibitors impact bond strength to dentin

Concern has been raised about the bonding of restorative procedures to an erosive lesion, given the change in organic and inorganic composition and structure of this substrate. This in vitro study evaluated the effect of erosive drinks and an enzyme inhibitor (2% chlorhexidine digluconate – 2% CHX) on bond strength to dentin. Sixty sound human third molars were selected, and the occlusal enamel was flattened, exposing the dentin surface. The specimens were randomly divided into three groups: AS-Artificial saliva (control group), RC- Regular Cola and ZC- Zero Cola. Twenty specimens were immersed in their respective solution for 1 minute, 3 times a day, over the course of 5 days. After acid etching and before bonding with Adper Single Bond 2, half of the samples of each group (n = 10) were treated with 2% CHX, whereas the other half (n = 10) were not, forming the control group (CONV). All the specimens were restored with Filtek Z250 composite resin filled in Tygon tubes (0.48 mm2), yielding six microcylinders for microshear bond strength testing. Three composite resin microcylinders of each specimen were tested after 1 month, and the remaining microcylinders were tested after 6 months. Failure modes were determined using a stereomicroscope (40x). The data were statistically analyzed by three-way ANOVA and Tukey tests (α = 0.05). Overall bonding was reduced after 6 months, regardless of treatment. The 2% CHX enhanced bond strength after 1 month only in the ZC group, and did not enhance bonding performance to demineralized dentin by erosive protocol after 6 months in any group.

Antisépticos: una visión química / The antiseptic ones: a chemical vision

En la actualidad, los antis‚pticos son ampliamente usados en hospitales y en otros centros de salud en una variedad de aplicaciones tópicas y de superficie. Los antis‚pticos son sustancias qu¡micas que se aplican sobre pie, heridas o cortes, con la finalidad de destruir o inhibir el crecimiento de microorganismos patógenos. En el presente trabajo se busca resaltar el conocimiento qu¡mico necesario, tales como la composición, concentración qu¡mica y modo de acción de tres de los antisépticos m sésencillos y de uso común como son el alcohol, agua oxigenada y el alcohol yodado.

Antiseptics are extensively used in hospitals and other health care settings for a variety of topical and hard-surface applications. There are chemicals that are applied on skin, wounds or cuts, in order to destroy or inhibit the growth of pathogenic microorganisms. This paper seeks to highlight the necessary chemical knowledge, such as composition, chemical concentration and mode of action of three antiseptics simple and commonly used, such as alcohol, peroxide and iodine alcohol.

Chlorhexidine does not increase immediate bond strength of etch-and-rinse adhesive to caries-affected dentin of primary and permanent teeth

The aim of this study was to evaluate the effect of 2% chlorhexidine digluconate (CHX) on immediate bond strength of etch-and-rinse adhesive to sound (SD) and caries-affected (CAD) primary dentin compared with permanent dentin. Flat dentin surfaces from 20 primary molars (Pri) and 20 permanent molars (Perm) were assigned to 8 experimental groups (n=5) according to tooth type (Pri or Perm), dentin condition (SD or CAD - pH-cycling for 14 days) and treatment (control - C or 60 s application of 2% CHX solution after acid etching - CHX). The bonding system (Adper Single Bond 2) was applied according to manufacturer's instructions followed by resin composite application (Filtek Z250). After 24 h water storage, specimens with cross-section area of 0.8 mm² were prepared for being tested under microtensile test (1 mm/min). Data were submitted to ANOVA and Tukey's post hoc test (α=0.05). Failure mode was evaluated using a stereomicroscope at ×400. Treatment with CHX did not result in higher bond strength values than no pre-treatment (C groups), independently of tooth type. Primary teeth and caries-affected dentin showed significantly lower (p<0.05) bond strength means compared with permanent teeth and sound dentin, respectively. Predominance of adhesive/mixed failure was observed for all groups. CHX did not influence the immediate bond strength to sound or caries-affected dentin of primary and permanent teeth.

O objetivo deste estudo foi avaliar o efeito do digluconato de clorexidina (CHX) a 2% na resistência de união imediata de adesivo convencional à dentina decídua hígida (H) e afetada pela lesão de cárie (CA) comparada à dentina permanente. Superfícies dentinárias planas de 20 molares decíduos (Dec) e 20 molares permanentes (Perm) foram divididas em 8 grupos experimentais (n=5) de acordo com o tipo de dente (Dec ou Perm), condição da dentina (H ou CA - ciclagem de pH por 14 dias) e tratamento (controle - C ou CHX - aplicação por 60 s de solução de digluconato de clorexidina a 2% após condicionamento ácido). O sistema adesivo (Adper Single Bond 2) foi aplicado de acordo com as instruções do fabricante, seguido da aplicação de resina composta (Filtek Z250). Após 24 h de armazenamento em água, espécimes com área de secção transversal de 0,8 mm² foram preparados para o teste de microtração (1 mm/min). Os dados foram submetidos à Análise de Variância e teste de Tukey para comparações múltiplas (α=0,05). Modo de falha foi avaliado usando estereomicroscópio com aumento de 400×. Tratamento com CHX não resultou em maiores valores de resistência de união do que sem pré-tratamento (C grupos), independente do tipo de dente. Dentes decíduos e dentina afetada apresentaram menores médias de resistência de união em comparação a dentes permanentes e dentina hígida, respectivamente (p<0,05). Predomínio de falha adesiva/mista foi observado para todos os grupos. CHX não influenciou a resistência de união imediata à dentina hígida ou afetada pela lesão de cárie de dentes decíduos e permanentes.

Influence of chlorhexidine application on longitudinal adhesive bond strength in deciduous teeth

The aim of this study was to evaluate the influence of applying 2 percent chlorhexidine for 30 seconds after phosphoric acid conditioning of dentin on the immediate and long-term bond strengths in deciduous teeth. The occlusal enamel was removed from 40 human sound deciduous molars, which were exfoliated by natural means, and the dentin was conditioned with 37 percent phosphoric acid for 15 seconds and washed with running water. The specimens were divided into two groups of 20 teeth. The test group received an application of 2 percent chlorhexidine for 30 seconds prior to a three-step etch-and-rinse adhesive system, whereas the control group received only the adhesive system. Three cylindrical restorations were made with a composite resin for each tooth. Ten teeth in each group were submitted to a microshear bond strength test after 24 hours, while the remaining teeth were stored in distilled water at 37 °C for 6 months before testing the microshear bond strength. The test group had a higher bond strength than did the control group after 6 months of storage. No statistical differences were found when groups with the same dentin treatment were compared at different times. Short applications of chlorhexidine at low concentrations prevent hybrid layer degradation and positively affect bond strength over time.

Effect of tetracycline on the bond performance of etch-and-rinse adhesives to dentin

This study evaluated the effect of modified tetracycline on the resin-dentin bond strength (µTBS), silver nitrate uptake (SNU) and solution homogeneity (SH) of two adhesives. Dentin surfaces were treated with phosphoric acid, rinsed off and either rewetted with water (control group - CO), 2 percent minocycline (MI), 2 percent doxycyline (DO) or 2 percent chlorhexidine (CH). Adhesive systems (Adper Single Bond 2 and Prime Bond NT) and composite were applied and light-polymerized. Specimens were sectioned to obtain bonded sticks (0.8 mm²) to test under tension at 0.5 mm/min. For SNU, specimens were immersed in silver nitrate and analyzed by EDX-SEM. SH was qualitatively analyzed after mixing the adhesives with different solvent-based solutions containing MI, DO and CH. Lower µTBS values were observed in the DO group compared with MI and CH (p = 0.01). Lower SNU was observed for MI and CH. The lowest µTBS for both adhesives was observed for the DO group (p = 0.01). Signs of phase separation were observed for DO with both adhesives. MI or CH used as rewetting solutions after acid etching did not affect the µTBS and hybrid layer quality.

Mechanism of oil-pulling therapy - In vitro study.

Background: Oil pulling has been used extensively as a traditional Indian folk remedy without scientific proof for many years for strengthening teeth, gums and jaws and to prevent decay, oral malodor, bleeding gums and dryness of throat and cracked lips. Aim: The aim of this study was to evaluate the antibacterial activity of sesame oil and lignans isolated from sesame oil on oral microorganisms and to check whether saponification or emulsification occurs during oil-pulling therapy. Materials and Methods: The in vitro study was carried out in three different phases: (1) Antibacterial activity of the lignans and sesame oil were tested by minimum inhibitory concentration assay by agar dilution method and agar well diffusion method, respectively. (2) Increase in free fatty acid level of oil and the quantity of sodium hydroxide (NaOH) used up in the titration are good indicators of saponification process. This was assessed using analytical tests for vegetable oils. (3) Swished oil was observed under light microscope to assess the status of the oil, presence of microorganisms, oral debris and foreign bodies. Results: Sesamin and sesamolin isolated from sesame oil did not have any antibacterial effect against oral microorganisms like Streptococcus mutans, Streptococcus mitis and Streptococcus viridans. Emulsification of sesame oil occurs during oil-pulling therapy. Increased consumption of NaOH in titration is a definite indication of a possible saponification process. Conclusion: The myth that the effect of oil-pulling therapy on oral health was just a placebo effect has been broken and there are clear indications of possible saponification and emulsification process, which enhances its mechanical cleaning action.

Evaluation of the antibacterial and physical properties of glass ionomer cements containing chlorhexidine and cetrimide: An in-vitro study.

Background: Incorporation of antibacterial agents frequently results in changes in the physical properties of restorative materials. Materials and Methods: This in-vitro study investigated the antibacterial and physical properties of Glass ionomer cement (GIC) with chlorhexidine and cetrimide, to determine the optimal concentration, for incorporation of these agents to obtain antibacterial GICs for use with the Atraumatic restorative treatment approach. This was assessed using the agar diffusion test. Chlorhexidine diacetate and cetrimide were added to Glass ionomer cement type-IX (GIC-FUJI IX) at 1 and 2% W/W ratio. The experimental GIC specimens were placed on agar plates inoculated with Lactobacillus casei, and the area of inhibition was calculated after 48 hours. Results: All the experimental GICs exhibited inhibition of bacteria, but the sizes were dependent on the concentration of the antibacterial agent. Incorporation of chlorhexidine diacetate and cetrimide, at 2%, significantly decreased the compressive strength, and the setting time was extended a little by the addition of any concentration of chlorhexidine and cetrimide. Conclusions: The present study demonstrated that experimental GICs containing chlorhexidine diacetate and cetrimide were effective in inhibiting bacteria associated with caries, and incorporation of 1% cetrimide was optimal to give the appropriate antibacterial and physical properties.

Influence of serum and necrotic soft tissue on the antimicrobial effects of intracanal medicaments

The purpose of this study was to investigate the influence of serum and necrotic soft tissue on the antimicrobial activity of intracanal medicaments. The medicaments tested were: calcium hydroxyde/glycerin paste, calcium hydroxide/chlorhexidine paste, calcium hydroxide/camphorated paramonochlorophenol/glycerin paste, and chlorhexidine/zinc oxide paste. Survival of Enterococcus faecalis and Candida albicans exposed to the medicaments tested in the presence or absence of serum or necrotic tissue was monitored in three in vitro experiments where samples for culturing were taken at different time periods. The overall results demonstrated that the antimicrobial activity of all intracanal medicaments tested was slowed down in the presence of necrotic tissue. Calcium hydroxide pastes in glycerin or chlorhexidine were significantly affected by serum. Of the medicaments tested in this study, the least affected was the calcium hydroxide/camphorated paramonochlorophenol/glycerin paste.

O objetivo deste estudo foi avaliar a influência do soro e de tecido mole necrosado na atividade antimicrobiana de medicamentos intra-canais. Os medicamentos testados foram pastas de hidróxido de cálcio/glicerina, hidróxido de cálcio/clorexidina, hidróxido de cálcio/paramonoclorofenol canforado/glicerina e clorexidina/óxido de zinco. A sobrevivência de Enterococcus faecalis e Candida albicans expostos aos medicamentos na presença ou ausência de soro ou tecido necrosado foi monitorada em três experimentos in vitro nos quais amostras para cultura foram avaliadas em diferentes períodos de tempo. No geral, os resultados demonstraram que a atividade antimicrobiana de todos os medicamentos testados foi retardada na presença de soro ou de tecido necrosado. As pastas de hidróxido de cálcio em glicerina ou clorexidina foram significativamente afetadas pelo soro. Dos medicamentos testados, o menos afetado foi a pasta de hidróxido de cálcio/paramonoclorofenol canforado/glicerina.

Concentration of hydrogen ions in several calcium hydroxide pastes over different periods of time

The purpose of this in vitro study was to measure the concentration of hydrogen ions (pH) of calcium hydroxide [(Ca(OH)2] pastes combined with different vehicles over 7 periods of time. The Ca(OH)2 was manipulated with the following vehicles: i: sterile water; ii: iodoform plus sterile water; iii: local anesthetics (Lydocaine 2 percent with 1: 100,000 epinephrine); iv: polyethyleneglycol; v: glycerin; vi: 2.0 percent chlorhexidine gel; vii: camphorated paramonochlorophenol (CMCP); viii: (CMCP) + glycerin; and ix: polyethyleneglycol plus CMCP. The pastes were made on a glass plate to toothpaste consistency and the pH was measured at the following times: 5 min, 1, 24, 48 h; 7, 14 and 28 days. The data were statistically analyzed (Kruskal-Wallis at p<0.05). At 5 min, 1 and 24 h, the pH of all tested pastes ranged from 13.05 to 11.16. At 48 h and 7 days the pH of all tested pastes ranged from 11.66 to 8.92. At 14 and 28 days almost all pastes had pH means lower than 10. In conclusion, the mean pH of all tested calcium hydroxide pastes decreased with the time. Pastes made with aqueous vehicles (especially with sterile water), followed by oily vehicles (especially with CMCP + glycerin), held the highest pH means over the periods of time tested.

O objetivo deste estudo in vitro foi mensurar a concentração de ions hidroxila (pH) de pastas de hidróxido de cálcio manipuladas com diversos veículos em 7 intervalos de tempo. As pastas foram manipuladas com os seguintes veículos: (i) água destilada estéril; (ii) idodofórmio + água destilada estéril; (iii) anestésico local (Lidocaína a 2 por cento com 1:100.000 epinefrina); (iv) polietilenoglicol (Calen); (v) glicerina; (vi) clorexidina gel 2 por cento; (vii) paramonoclorofenol canforado (PMCC); (viii) PMCC + glicerina; e (ix) PMCC + polietilenoglicol (Calen PMCC). As pastas foram manipuladas em consistência de pasta de dente e os pH mensurados 5 min; 1, 24, 48 h; 7, 14 e 28 dias após manipulação. Os resultados foram analisados estatisticamente através do teste Kruskal-Wallis (p<0,05). Aos 5 min, 1 e 2 h após manipulação o pH de todas as pastas ficou entre 13.05 e 11.16. Aos 48 h e 7 dias após a manipulação, o pH de todas as pastas testadas variou de 11.66 a 8.92. Aos 14 e 28 dias, quase todas as pastas mostraram pH menor que 10. Concluiu-se que o pH de todas as pastas hidróxido de cálcio decresceram de acordo com o tempo. Pastas feitas com veículos aquosos (especialmente com água destilada), seguida de veículos oleosos (especialmente com CMCP + glicerina) mantiveram as maiores médias de pH durante os períodos testados.

Freeze-drying as an alternative method of human sclera preservation / Liofilização como alternativa à preservação de esclera humana

PURPOSE: To compare the effect of preserving sclera samples in either 95 percent ethanol or freeze-dried. METHODS: Ninety-six samples of human sclera were studied. Half of them were freeze-dried and half preserved in 95 percent ethanol. Preservation periods of 18, 45, 90 or 174 days were studied. Automated immunostaining was carried out in the Ventana BenchMarkR LT platform using collagen 1 and fibronectin antibodies. Histological staining was also performed with hematoxilin-eosin and Masson trichrome. Samples were classified according to the degree of collagen fiber parallelism (0-2), intensity of Masson staining (0-2), and the expression of both antibodies (0-3). Friedman and Wilcoxon tests were applied to compare preservation methods and p-values below 0.05 were considered to ensure statistical significance. RESULTS: Relevant results were found in three situations: (i) Friedman's test showed better collagen fiber integrity in the freeze-dried group rehydrated after 174-days as compared to the 90-day group; (ii) Wilcoxon's test showed better collagen fiber integrity in the freeze-dried group after 18 and 174 days as compared to the ethanol group; (iii) the freeze-dried group disclosed higher immunohistochemical expression for COL-1 antibody in the sclera samples rehydrated after 45, 90 and 174 days as compared to the ones rehydrated after 18 days. CONCLUSION: Histological and immunohistochemical analysis showed freeze-drying to be a superior method for sclera preservation as compared to 95 percent ethanol. This technique provides an easy method to manipulate tissue, with longer shelf life, and storage at room temperature.

OBJETIVO: Comparar dois métodos de preservação de esclera humana, liofilização e álcool 95 por cento, em diferentes períodos de tempo. MÉTODOS: Foram avaliados 96 fragmentos de seis escleras humanas. Metade das amostras foi submetida ao processo de liofilização e metade conservada em álcool 95 por cento. Dois fragmentos de cada grupo foram avaliados pelas colorações de hematoxilina-eosina e tricrômio de Masson e submetidos a técnica de imuno-histoquímica para os anticorpos fibronectina e colágeno 1, após 18, 45, 90 e 174 dias de preservação. Os espécimens foram classificados de acordo com o paralelismo (PF:0-2) e integridade (IF:0-1) das fibras de colágeno e expressão imuno-histoquímica para os anticorpos fibronectina (FIB:0-3) e colágeno 1 (COL-1:0-3). A análise estatística foi realizada por meio dos testes de Friedman e Wilcoxon e o valor de p menor que 0,05 foi considerado estatisticamente significante. RESULTADOS: Verificaram-se diferenças significantes em três situações: (i) maior integridade das fibras de colágeno das escleras liofilizadas após 174 dias quando comparado aos 90 dias; (ii) maior expressão imuno-histoquímica para o anticorpo COL-1 nas amostras de escleras liofilizadas após os 18 dias iniciais de preservação; (iii) maior integridade das fibras de colágeno das escleras liofilizadas após 18 e 174 dias quando comparado às escleras preservadas em álcool. CONCLUSÕES: A preservação de tecido escleral por liofilização mostrou-se técnica tão eficaz quanto a preservação em álcool, apresentado vantagem quando considerada a integridade das fibras de colágeno. A liofilização mostra-se benéfica por permitir a estocagem do tecido em temperatura ambiente e com prazo de validade estendido.

Evaluation of the antimicrobial activity of each component in Grossman's sealer

Os autores estudaram a atividade antimicrobiana do cimento de Grossman e de seus componentes sobre 13 diferentes cepas pelo método de difusão de poço em camada dupla. Os resultados revelaram que o cimento de Grossman apresentou atividade antimicrobiana contra todas as cepas utilizadas. Dos componentes do cimento, o tetraborato de sódio foi o que apresentou maior atividade antimicrobiana, tanto por tipo como por tamanho do halo e aro de inibição. O pó do cimento, o breu e o eugenol apresentaram atividades semelhantes, sendo que eles não tiveram ação sobre P. aeruginosa e C. albicans e, dos três componentes, somente o eugenol teve ação sobre E coli. O óxido de zinco somente teve ação sobre S. sobrinus e E. coli. O sulfato de bário e o subcarbonato de bismuto não tiveram nenhuma ação antimicrobiana.