Development and Physicochemical Characterization of Eugenia brejoensis Essential Oil-Doped Dental Adhesives with Antimicrobial Action towards Streptococcus mutans.

Dental caries is a multifactorial, biofilm-dependent infectious disease that develops when detrimental changes occur in the oral cavity microenvironment. The antimicrobial and antivirulence properties of the essential oil obtained from the leaves of Eugenia brejoensis Mazine (EBEO) have been reported against Gram-positive and Gram-negative bacteria. Herein, the antimicrobial action of EBEO towards Streptococcus mutans is reported, along with the development and characterization of dental adhesives doped with. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of EBEO were determined against S. mutans, while its toxicity was analyze using Tenebrio molitor larvae. EBEO (MIC and 10×MIC) was incorporated into the Ambar Advanced Polymerization System® (Ambar APS), a two-step total-etch adhesive system (FGM Dental Group), and the antibiofilm action was evaluated. The reflective strength, modulus of elasticity, degree of conversion, and maximum rate of polymerization of each adhesive were also determined. The MIC and MBC values of EBEO against S. mutans were 62.5 µg/mL. The tested concentrations of EBEO were non-toxic to T. molitor larvae. The formation of S. mutans biofilms was significantly inhibited by EBEO and EBEO-coated resin discs (p < 0.05). Importantly, EBEO incorporation did not affect the mechanical and physicochemical properties in relation to oil-free adhesive version. EBEO showed strong antibacterial and antibiofilm activity against S. mutans, no toxicity effect against T. molitor larvae, and did not jeopardize the physical-chemical properties tested.

Hydroalcoholic Leaf Extract of Punica granatum, alone and in Combination with Calcium Hydroxide, Is Effective against Mono- and Polymicrobial Biofilms of Enterococcus faecalis and Candida albicans.

Failures in endodontic treatments are mostly associated with the difficulty in eradicating microbes of the root canal system, highlighting the need to develop novel effective antimicrobials. Punica granatum (pomegranate) leaf hydroalcoholic extract may be a potential alternative in canal dressing, owing to its antimicrobial properties. The objective of this study was to evaluate the antimicrobial activity of hydroalcoholic leaf extract of Punica granatum (HEPg) alone or in combination with calcium hydroxide (Ca(OH)2) against Enterococcus faecalis and Candida albicans in isolation and in mono- and polymicrobial biofilms. Microdilution tests in broth and assays for inhibition of biofilm formation were carried out to evaluate the antimicrobial properties of HEPg and HEPg + Ca(OH)2 against Enterococcus faecalis and Candida albicans. The cytotoxicity of HEPg in HaCaT cells was evaluated by MTT assay. HEPg and HEPg + Ca(OH)2 exerted significant antimicrobial activity against planktonic cells and mono- and polymicrobial biofilms. The combination of Punica granatum extract with Ca(OH)2 appears to be a promising alternative in endodontic treatments, which could be tested in vivo to confirm the efficacy of this mixture in disinfecting root canal systems.

Bonding Efficacy of Universal Adhesives to Fluorotic Enamel after Pre-conditioning with EDTA.

PURPOSE: To compare the effect of active pre-conditioning with 17% ethylenediaminetetraacetic acid (EDTA) vs 37% phosphoric acid (PA) on the resin-enamel microshear bond strength (µSBS), enamel-etching pattern, and in situ degree of conversion (in situ DC) of four universal adhesives on sound and fluorotic enamel. MATERIAL AND METHODS: In this study, 448 extracted human molars (224 without fluorosis and 224 with fluorosis) were sectioned into four parts and divided into 16 experimental groups based on the enamel surface (sound or fluorotic enamel), adhesive (Clearfil Universal Bond [CUB], Futurabond U [FBU], iBond Universal [IBU], or Scotchbond Universal [SBU]), and enamel conditioning agent (PA or EDTA). The specimens were stored for 24 h and tested under shear stress at 1.0 mm/min to determine the µSBS. The adhesive-enamel interfaces were evaluated for in situ DC using micro-Raman spectroscopy. The enamel-etching pattern was evaluated using a scanning electron microscope. The µSBS and in situ DC data were analyzed separately using three-way ANOVA and Tukey's post-hoc test (a = 0.05). RESULTS: Sound enamel showed higher µSBS and in situ DC compared to fluorotic enamel (p < 0.05). However, no significant difference was observed for µSBS, in situ DC (p > 0.05), or etching patterns when PA and EDTA etching were compared in sound and fluorotic enamel. Moreover, CUB and SBU showed higher mean µSBS than did FBU and IBU in both sound and fluorotic enamel (p < 0.05). CONCLUSIONS: Compared to PA, active pre-conditioning with EDTA showed similar µSBS and enamel etching patterns for all the adhesives in fluorotic enamel, without compromising the in situ DC.

Modulation of Streptococcus mutans virulence by dental adhesives containing anti-caries agents.

OBJECTIVES: The aim of this in vitro study was to analyze the effect of the incorporation of two anti-caries agents into dental adhesives on the reduction of the virulence of Streptococcus mutans and on the adhesion to dentin. METHODS: Apigenin (1mM) and tt-Farnesol (5mM) were added separately and in combination to a self-etch adhesive (CS3 - Clearfil S3 Bond Plus) and to an each-and-rinse adhesive (OPT - OptiBond S). Biofilm of S. mutans was grown on adhesive-coated hydroxyapatite disks for 115h and bacterial viability, dry-weight, alkali soluble, water soluble, intracellular polysaccharides and protein were quantified. Bond strength and dentin-adhesive interface were performed to analyze the effects of the incorporation on the physical properties and to identify changes in hybrid layer morphology. RESULTS: Addition of Apigenin and Apigenin+tt-Farnesol to CS3, and Apigenin or tt-Farnesol to OPT reduced the dry-weight of S. mutans biofilm. Insoluble polysaccharide decreased with the addition of Apigenin to CS3 and tt-Farnesol to OPT. Intracellular polysaccharide decreased with addition of Apigenin and Apigenin+tt-Farnesol to CS3. No changes in dentin bond strength, resin-dentin interfacial morphology, total amount of protein and soluble polysaccharide were observed with the additions. SIGNIFICANCE: Biofilms that are less cariogenic around dental restorations could decrease secondary caries formation; in addition, the reduction of virulence of S. mutans without necessarily killing the microorganism is more unlikely to induce antimicrobial resistance.

Antibacterial activity of diacetylcurcumin against Staphylococcus aureus results in decreased biofilm and cellular adhesion.

PURPOSE: Staphylococcus aureus infections have contributed to the global healthcare burden, particularly with regard to hospital-acquired meticillin-resistant S. aureus (MRSA) infections. METHODOLOGY: This study describes the antibacterial activity of diacetylcurcumin (DAC) against meticillin-susceptible S. aureus/MRSA biofilm formation, survival, metabolic activity and structure; its ability to prevent bacterial adhesion to human cells; and toxicity in Galleria mellonella larvae. RESULTS: DAC showed excellent antibacterial activity, with MIC ranging between 17.3 and 34.6 µmol l-1, and minimum bactericidal concentration ranging between 69 and 277 µmol l-1. It significantly reduced bacterial biofilm survival - by 22-63 % (at MIC, 10×MIC or 100×MIC) as compared to the 25-42 % reduction by vancomycin (P<0.0001) - and severely affected biofilm cell structures, leading to damaged architecture and the formation of amorphous cell clusters. Treatment with DAC (MIC/4) decreased bacterial adhesion to HaCaT keratinocytes from 1 to 5 h (P<0.0001). Finally, DAC demonstrated low toxicity in G. mellonella at its effective anti-biofilm concentrations. CONCLUSION: These findings open new avenues for the study of this curcumin derivative as an excellent prototype with anti-MRSA activity.

The Effect of Essential Oils and Bioactive Fractions on Streptococcus mutans and Candida albicans Biofilms: A Confocal Analysis.

The essential oils (EO) and bioactive fractions (BF) from Aloysia gratissima, Baccharis dracunculifolia, Coriandrum sativum, Cyperus articulatus, and Lippia sidoides were proven to have strong antimicrobial activity on planktonic microorganisms; however, little is known about their effects on the morphology or viability of oral biofilms. Previously, we determined the EO/fractions with the best antimicrobial activity against Streptococcus mutans and Candida spp. In this report, we used a confocal analysis to investigate the effect of these EO and BF on the morphology of S. mutans biofilms (thickness, biovolume, and architecture) and on the metabolic viability of C. albicans biofilms. The analysis of intact treated S. mutans biofilms showed no statistical difference for thickness in all groups compared to the control. However, a significant reduction in the biovolume of extracellular polysaccharides and bacteria was observed for A. gratissima and L. sidoides groups, indicating that these BF disrupt biofilm integrity and may have created porosity in the biofilm. This phenomenon could potentially result in a weakened structure and affect biofilm dynamics. Finally, C. sativum EO drastically affected C. albicans viability when compared to the control. These results highlight the promising antimicrobial activity of these plant species and support future translational research on the treatment of dental caries and oral candidiasis.

Apolar Bioactive Fraction of Melipona scutellaris Geopropolis on Streptococcus mutans Biofilm.

The aim of this study was to evaluate the influence of the bioactive nonpolar fraction of geopropolis on Streptococcus mutans biofilm. The ethanolic extract of Melipona scutellaris geopropolis was subjected to a liquid-liquid partition, thus obtaining the bioactive hexane fraction (HF) possessing antimicrobial activity. The effects of HF on S. mutans UA159 biofilms generated on saliva-coated hydroxyapatite discs were analyzed by inhibition of formation, killing assay, and glycolytic pH-drop assays. Furthermore, biofilms treated with vehicle control and HF were analyzed by scanning electron microscopy (SEM). HF at 250 µ g/mL and 400 µ g/mL caused 38% and 53% reduction in the biomass of biofilm, respectively, when compared to vehicle control (P < 0.05) subsequently observed at SEM images, and this reduction was noticed in the amounts of extracellular alkali-soluble glucans, intracellular iodophilic polysaccharides, and proteins. In addition, the S. mutans viability (killing assay) and acid production by glycolytic pH drop were not affected (P > 0.05). In conclusion, the bioactive HF of geopropolis was promising to control the S. mutans biofilm formation, without affecting the microbial population but interfering with its structure by reducing the biochemical content of biofilm matrix.

Antimicrobial Activity of Essential Oils against Streptococcus mutans and their Antiproliferative Effects.

This study aimed to evaluate the activity of essential oils (EOs) against Streptococcus mutans biofilm by chemically characterizing their fractions responsible for biological and antiproliferative activity. Twenty EO were obtained by hydrodistillation and submitted to the antimicrobial assay (minimum inhibitory (MIC) and bactericidal (MBC) concentrations) against S. mutans UA159. Thin-layer chromatography and gas chromatography/mass spectrometry were used for phytochemical analyses. EOs were selected according to predetermined criteria and fractionated using dry column; the resulting fractions were assessed by MIC and MBC, selected as active fractions, and evaluated against S. mutans biofilm. Biofilms formed were examined using scanning electron microscopy. Selected EOs and their selected active fractions were evaluated for their antiproliferative activity against keratinocytes and seven human tumor cell lines. MIC and MBC values obtained for EO and their active fractions showed strong antimicrobial activity. Chemical analyses mainly showed the presence of terpenes. The selected active fractions inhibited S. mutans biofilm formation (P < 0.05) did not affect glycolytic pH drop and were inactive against keratinocytes, normal cell line. In conclusion, EO showed activity at low concentrations, and their selected active fractions were also effective against biofilm formed by S. mutans and human tumor cell lines.

Reconstrução estética utilizando prótese adesiva como forma de reabilitação oral em serviço público - relato de caso / Aesthetic reconstruction using adhesive prosthesis for oral rehabilitation in public service - case report

Objetivo: Relatar um caso clínico sobre uso de prótese adesiva para reabilitação bucal em serviço público odntológico. Métodos: Foram confeccionados pônticos da prótese adesiva com resina composta reforçada por fibra de vidro para substituir dentes ântero-superiores, perdidos em paciente atendida na clínica odontológica da escola Sotero dos Reis, integrante do Programa Saúde na Escola. Resultados: Dentro das limitações de uma instituição pública, foi possível realizar um procedimento clínico, houve o restabelecimento da função, estética e a inclusão social da paciente.

Objective: To present a clinical case report about using adhesive prosthesis for mouth rehabilitation in public dental assistance. Methods: Pontics of adhesive prosthesis were placed in patient utilizing direct resin-bonded fibrereinforced composite for maxillary central anterior teeth replacement. The study was held at the dental clinics of Sotero dos Reis School Health Program. Results: Within the limitations of public service, it was possible to perform oral rehabilitation satisfactory. Conclusion: This clinical procedure was viewed as a simple means of allowing the function, aesthetic and social patient's inclusion.