Aline Rogéria Freire de Castilho

In vitro and in vivo investigation of the biological and mechanical behaviour of resin-modified glass-ionomer cement containing chlorhexidine

OBJECTIVES To evaluate: (1) the in vitro antibacterial, cytotoxic and mechanical properties of a resin-modified glass ionomer cement (RMGIC) containing different concentrations of chlorhexidine (CHX) and (2) the in vivo microbiologic action of the best concentration of CHX associated with the RMGIC applied on remaining dentine after indirect pulp treatment (IPT). METHODS For the in vitro studies, RMGIC was associated with 0.2, 0.5, 1.25 and 2.5% CHX. Microbiologic evaluation consisted of an agar diffusion test on cariogenic bacteria for 24h. Odontoblast-like cell metabolism and morphology analyses measured the cytotoxic effects of the RMGIC groups after 24h. The same groups were submitted to compressive and diametral tensile strength. The in vivo treatment consisted of IPT using an RMGIC associated with the best CHX concentration. Clinical and microbiologic evaluations were performed before and after 3 months. RESULTS The use of 1.25% CHX significantly improved the antibacterial effects of the evaluated RMGIC, without causing any detrimental effects to the odontoblast-like cells and on the mechanical properties. This RMGIC and CHX combination completely eliminated mutans streptococci after 3 months of IPT. CONCLUSION The RMGIC and 1.25% CHX mixture showed great biological and mechanical behaviour and could be a good treatment against caries progression. CLINICAL SIGNIFICANCE The association of CHX with a liner RMGIC opens a new perspective for arresting residual caries after IPT.

Mechanical and biological characterization of resin-modified glass-ionomer cement containing doxycycline hyclate

OBJECTIVES To characterize the mechanical and biological properties of a resin-modified glass ionomer cement (RMGIC) containing doxycycline hyclate. METHODS The antibacterial effect of RMGIC containing 1.5, 3.0 and 4.5% doxycycline hyclate was assessed using two experiments - agar diffusion test for 24h and biofilm assay for 24h and 7 days - against some cariogenic bacteria. Briefly, base layers of BHI agar and 300μL of each inoculum were prepared in Petri dishes with 6 wells that were completely filled with materials. After 24h incubation, zones of bacterial growth inhibition were measured using a digital caliper. Biofilm assays were conducted using RMGIC specimens immersed in 24-well plates containing the inoculum in BHI broth. After 24h and 7 days, each specimen were removed, vortexed and the suspension diluted and inoculated in BHI plates for subsequent bacterial counting. Cytotoxicity tests used 50 specimens made in sterilized metal molds, including Vitrebond as positive control. Extracts from every specimen were applied on the MDPC-23 odontoblast-like cells for 24h. The MTT assay and SEM evaluation determined cell metabolism and morphology, respectively. 80 cylindrical specimens were made from the previously cited groups, and were submitted to testing with a universal testing machine (Instron 4411) using a crosshead speed of 1.0mm/min for compressive strength and 0.5mm/min for diametral tensile strength, respectively. Data from antibacterial and cytotoxic effects, and mechanical properties were submitted to appropriated statistical tests. RESULTS All tested groups showed growth inhibition of all tested strains (p<0.05) in 24h for both microbiological tests, but only 4.5% doxycycline have antibacterial effect after 7 days. None of doxycycline concentrations caused toxic effect to the MDPC-23 cells or presenting alterations to mechanical properties. CONCLUSION The incorporation of up to 4.5% doxycycline hyclate into RMGIC inhibits important oral microorganisms, without modifying biological and mechanical characteristics of the dental material, suggesting a new alternative for the treatment of dental caries.

Protective effect of experimental mouthrinses containing NaF and TiF4 on dentin erosive loss in vitro

Objective This in vitro study assessed the anti-erosive effect of experimental mouthrinses containing TiF4 and NaF on dentin erosive loss. Material and Methods Bovine dentin specimens were randomly allocated into the groups (n=15): 1) SnCl2/NaF/AmF (Erosion Protection®/GABA, pH 4.5, positive control); 2) experimental solution with 0.0815% TiF4 (pH 2.5); 3) 0.105% NaF (pH 4.5); 4) 0.042% NaF+0.049% TiF4 (pH 4.4); 5) 0.063% NaF+0.036% TiF4 (pH 4.5); 6) no treatment (negative control). Each specimen was cyclically demineralized (Sprite Zero, pH 2.6, 4x90 s/day) and exposed to artificial saliva between the erosive challenges for 7 days. The treatment with the fluoride solutions was done 2x60 s/day, immediately after the first and the last erosive challenges of the day. Dentin erosive loss was measured by profilometry (μm). The data were analyzed using Kruskal Wallis/Dunn tests (p<0.05). Results Mouthrinses containing TiF4 or Sn/F were able to show some protective effect against dentin erosive loss compared to negative control. The best anti-erosive effect was found for experimental solution containing 0.0815% TiF4 (100% reduction in dentin loss), followed by 0.042% NaF+0.049% TiF4 (58.3%), SnCl2/NaF/AmF (52%) and 0.063% NaF+0.036% TiF4 (40%). NaF solution (13.3%) did not significantly differ from control. Conclusion The daily application of experimental mouthrinse containing TiF4 and NaF has the ability to reduce dentin erosion, as well as Erosion Protection® and TiF4 alone.

Avaliação da incidência de cárie em pacientes com síndrome de Down após sua inserção em um programa preventivo

The objective of this work was to verify the incidence of dental caries by means of the CPO-D, CPO-S, ceo-d and ceo-s indexes in patients with Down syndrome regularly enrolled in a preventive program. Twenty four Down syndrome patients of both sexes age range of one to 48 years were examined. The prevalence (initial experimental situation) and incidence (final experimental situation) of dental caries were verified using of the initial and final CPO-D, CPO-S, ceo-d and ceo-s indexes of the participants. From 24 individual examined, 10 (42.0%) were free of caries. The prevalence of dental caries showed values of CPO-D= 2.33; CPO-S= 3.60; ceo-d= 1.75 e ceo-s= 2.80; while the incidence of caries showed values of 2.33; 3.80; 1.10 e 1.90, respectively. Down syndrome individuals evaluated in this study presented low level of caries and small incidence of new lesions, emphasizing the importance of the maintenance of these patients at preventive programs.

Consequences and treatment after multiple avulsions of deciduous teeth – a case report

The prevalence of traumatisms with avulsion of deciduous teeth varies from 7 to 13%, usually involving one tooth only. In this case report, a trauma with multiple losses of deciduous teeth and laceration of soft tissue is described as a consequence of a horse backward kick. After suture of extra-oral tissue wounds and application of anti-tetanic vaccine, the patient was referred to the emergency ambulatory of Sacred Heart University. Treatment procedures included radiographic analysis, removal of bone fragments and of a residual root of the tooth involved, remodeling and suture of intra-oral tissues. The patient was observed periodically and after full recovery, a prosthetic appliance was installed for functional and esthetic rehabilitation.

Doxycycline-containing glass ionomer cement for arresting residual caries: an in vitro study and a pilot trial

Abstract In a previous study, we demonstrated that the incorporation of doxycycline hyclate (DOX) into resin-modified glass ionomer cement (RMGIC) inhibited important cariogenic microorganisms, without modifying its biological and mechanical characteristics. In this study, we keep focused on the effect of that experimental material as a potential therapy for arresting residual caries by analyzing other in vitro properties and conducting a pilot clinical trial assessing the in vivo effect of DOX-containing RMGIC on residual mutans streptococci after partial carious removal in primary molars. Specimens of the groups RMGIC (control); RMGIC + 1.5% DOX; RMGIC + 3% DOX; and RMGIC + 4.5% DOX were made to evaluate the effect of DOX incorporation on surface microhardness and fluoride release of RMGIC and against biofilm of Streptococcus mutans. Clinical intervention consisted of partial caries removal comparing RMGIC and RMGIC + 4.5% DOX as lining materials. After 3 months, clinical and microbiologic evaluations were performed. Data were submitted to ANOVA/Tukey or Wilcoxon/Mann-Whitney set as α=0.05. Fluoride release and surface microhardness was not influenced by the incorporation of DOX (p>0.05). There was a significant reduction of S. mutans biofilm over the material surface with the increase of DOX concentration. After clinical trial, the remaining dentin was hard and dry. Additionally, mutans streptococci were completely eliminated after 3 months of treatment with RMGIC + 4.5% DOX. The incorporation of DOX provided better antibiofilm effect, without jeopardizing fluoride release and surface microhardness of RMGIC. This combination also improved the in vivo shortterm microbiological effect of RMGIC after partial caries removal.

Antibacterial activity against cariogenic bacteria and cytotoxic and genotoxic potential of Anacardium occidentale L. and Anadenanthera macrocarpa (Benth.) Brenan extracts

OBJECTIVES The present study aimed to assess the antibacterial activity against bacteria with cariogenic relevance, toxic and genotoxic potential of the plants Anacardium occidentale L. and Anadenanthera macrocarpa (Benth.) Bernam. DESIGN Using a microdilution technique, the extracts were submitted to minimum inhibitory concentration (MIC) testing against Streptococcus mitis (ATCC 903), Streptococcus mutans (ATCC 25175), Streptococcus oralis (ATCC 10557), Streptococcus salivarius (ATCC 7073), Streptococcus sanguinis (ATCC 15300) and Streptococcus sobrinus (ATCC 27609). The toxicity of the extracts was then verified against eukaryotic cells. Additionally, a micronucleus assay was performed to investigate the potential mutagenic effects of the extracts on rat erythrocytes. The Students t-test, Bonferroni test, and one-way ANOVA followed by Tukeys tests were used for statistical analysis, at a significance level of 5%. RESULTS While the A. occidentale extract was able to inhibit all of the tested strains, with S. mutans and S. mitis being the most susceptible to that extract́s action, the A. macrocarpa did not show antimicrobial activity. Interestingly, the hemolytic, oxidant and antioxidant activities were slightly observed for either extract, even at high concentrations (1000mg/mL). The micronucleus assay showed no significant changes in the cells exposed to the extracts. CONCLUSION The A. occidentale extract has potential as an antimicrobial agent with low eukaryotic cell toxicity or mutagenic activity. The A. macrocarpa extract, although absent of antibacterial activity might as well be a safe and effective phytotherapeutic alternative.

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.