Hevea brasiliensis rubber particles' fluid interfaces reveal size impact on early coagulation steps.

Natural rubber originates from the coagulation of rubber particles (RP) from Hevea brasiliensis latex. The size distribution of Hevea RP is bimodal with the presence of small rubber particles (SRP) and large rubber particles (LRP). This study aims at getting a better understanding of the early coagulation steps of Hevea RP taking into account the particle size. SRP and LRP were obtained by centrifugation of freshly tapped ammonia-free latex from RRIM600 clone. Size and zeta potential measurements showed that both RP fractions were efficiently separated and stable in basic buffer. SRP and LRP dispersions were placed in a Langmuir trough and RP were let to adsorb at the air-liquid interface to form interfacial films. Surface tension and ellipsometry indicate that the formation kinetics and the stabilization of the film at the air-liquid interface are faster for SRP than LRP. Moreover, the arrangement of RP at the interface differs between SRP and LRP, as shown by Brewster angle microscopy, atomic force microscopy and confocal laser scanning microscopy. First, the RP membrane and cis-1,4-polyisoprene core spread at the air-liquid interface before clustering. Then, while the SRP fuse, the LRP keep their structure in individual particles in floating aggregate. The role of the non-isoprene molecules on the different organization of SRP and LRP films is discussed, the one of the two major RP proteins, SRPP1 (Small Rubber Particle Protein) and Rubber Elongation Factor (REF1) in the early coagulation steps.

Assessment of multispecies biofilm growth on root canal dentin under different radiation therapy regimens.

OBJECTIVES: To assess the growth of a multispecies biofilm on root canal dentin under different radiotherapy regimens. MATERIALS AND METHODS: Sixty-three human root dentin cylinders were distributed into six groups. In three groups, no biofilm was formed (n = 3): NoRT) non-irradiated dentin; RT55) 55 Gy; and RT70) 70 Gy. In the other three groups (n = 18), a 21-day multispecies biofilm (Enterococcus faecalis, Streptococcus mutans, and Candida albicans) was formed in the canal: NoRT + Bio) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm. The biofilm was quantified (CFUs/mL). Biofilm microstructure was assessed under SEM. Microbial penetration into dentinal tubules was assessed under CLSM. For the biofilm biomass and dentin microhardness pre- and after biofilm growth assessments, 45 bovine dentin specimens were distributed into three groups (n = 15): NoRT) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm. RESULTS: Irradiated specimens (70 Gy) had higher quantity of microorganisms than non-irradiated (p = .010). There was gradual increase in biofilm biomass from non-irradiated to 55 Gy and 70 Gy (p < .001). Irradiated specimens had greater reduction in microhardness after biofilm growth. Irradiated dentin led to the growth of a more complex and irregular biofilm. There was microbial penetration into the dentinal tubules, regardless of the radiation regimen. CONCLUSION: Radiotherapy increased the number of microorganisms and biofilm biomass and reduced dentin microhardness. Microbial penetration into dentinal tubules was noticeable. CLINICAL RELEVANCE: Cumulative and potentially irreversible side effects of radiotherapy affect biofilm growth on root dentin. These changes could compromise the success of endodontic treatment in oncological patients undergoing head and neck radiotherapy.

Effect of obturation technique on dentinal tubule penetration of two tricalcium silicate-based sealers: Ex vivo study.

The aim of this study was to assess by confocal laser microscope the depth of dentinal tubule penetration of two tricalcium silicate-based sealers promoted by two obturation techniques in curved canals compared with AHPlus. One hundred and twenty canals were divided into six groups (n = 20): BCSC-Bio-C Sealer (BC) and single-cone technique (SC); BCCW-BC and continuous condensation wave (CW); TFSC-Total Fill (TF) and SC; TFCW-TF and CW; AHSC-AH Plus (AH) and SC; AHCW-AH and CW. Data were analysed using the three-way ANOVA and Tukey's test (α = 5%). Penetration depth was significantly greater for TFCW than TFSC and greater for AHCW than AHSC (p < 0.05). There was no significant difference between BCCW and BCSC (p > 0.05). The penetration of TF was significantly greater (p < 0.05). The CW technique promoted greater intratubular penetration, except for the BC sealer.

Effect of different obturation techniques on sealer penetration into dentinal tubules after final irrigation with XP-Endo finisher file

The first objective of this research was to evaluate the effectiveness of XP-Endo Finisher on dentinal tubule penetration of irrigation solution using confocal laser scanning microscopy. The main purpose of this research was to compare the effect of cold lateral condensation, continuous wave obturation and core-carrier based techniques on sealer penetration. Sixty mandibular premolars were prepared and allocated into two experimental groups (n=30) as the final irrigation technique and obturation technique experiment. In the final irrigation technique experiment, final irrigation was performed with XP-Endo Finisher, passive ultrasonic irrigation (PUI) and conventional needle irrigation (CNI) (n=10). The roots in the obturation technique experiment were also assigned into 3 groups and obturated with cold lateral condensation, continuous-wave obturation and core-carrier techniques (n=10). The most effective activation method, which emerged as a result of the first part of this study, was used as the final irrigation method in the obturation technique experiment. Then, all roots were sectioned in 1-mm-thick slices at 3mm from the apex for scanning. In terms of depth and percentage of material penetration, CNI exhibited significantly the lowest values and no significant difference was found between others. Also, there was no significant difference among obturation methods. In conclusion, XP-Endo Finisher and PUI are more effective than CNI on irrigant penetration. Sealer penetration into dentinal tubules is independent of obturation techniques.

El objetivo principal de esta investigación fue evaluar la eficacia de XP- Endo Finisher en la penetración de la solución de irrigación en los túbulos dentinarios mediante microscopía de láser confocal. Se prepararon sesenta premolares mandibulares y se distribuyeron en dos grupos experimentales (n=30) según el tipo de método de evaluación utilizado. En el experimento de la técnica de irrigación final, la irrigación final se realizó con XP-Endo Finisher, irrigación ultrasónica pasiva (PUI) e irrigación con aguja convencional (CNI) (n=10). Las raíces en el experimento de la técnica de obturación también se asignaron en 3 grupos y se obturaron con técnicas de condensación lateral fría, obturación de onda continua y portador de núcleo (n=10). El método de activación más eficaz, que surgió como resultado de la primera parte de este estudio, se utilizó como método de irrigación final en el experimento de la técnica de obturación. Luego, todas las raíces se seccionaron en muestras de 1mm de espesor. En términos de profundidad y porcentaje de penetración del material, CNI exhibió significativamente los valores más bajos y no se encontraron diferencias significativas entre los demás. Además, no hubo diferencias significativas entre los métodos de obturación. En conclusión, XP-Endo Finisher y PUI son más efectivos que CNI en la penetración del irrigante. La penetración del sellador en los túbulos dentinarios es independiente de las técnicas de obturación.

Effect of ultrasonic agitation on marginal adaptation, intratubular penetration and bonding strength of calcium-silicate-based endodontic sealers: A laboratory study.

The effect of ultrasonic activation (UA) on marginal adaptation, intratubular penetration and bond strength provided by three calcium silicate-based sealers was evaluated. Ninety-six distobuccal root canals of maxillary molars were randomly divided into 8 groups (n = 12) according to the sealer and UA application: EndoSequence BC sealer (ESBC), Sealer Plus BC (SPBC) and Bio-C Sealer (BCS), using AH Plus (AH) as a control group. The specimens were sectioned at 2, 4 and 6 mm from the apex. The data were statistically analysed using Kruskall-Wallis, Dunn, Mann-Whitney and chi-squared tests. UA improved the marginal adaptation of ESBC (6 mm), SPBC (all levels), BCS (2/4 mm) and AH (4 mm) (p < 0.05); the bond strength of SBPC (2 mm) and BCS (6 mm) were also improved (p < 0.05). The UA of endodontic silicate-based sealers improved the marginal adaptation in all levels and the bond strength of SBPC and BCS sealer.

Acute toxicity of titanium dioxide microparticles in Artemia sp. nauplii instar I and II.

In this study, the toxicity effects of titanium dioxide (MTiO2 ) microparticles on Artemia sp. nauplii instar I and II between 24 and 48 h was evaluated. The MTiO2 were characterized using different microscopy techniques. MTiO2 rutile was used in toxicity tests at concentration of 12.5, 25, 50, and 100 ppm. No toxicity was observed in Artemia sp. nauplii instar I at the time of 24 and 48 h. However, Artemia sp. nauplii instar II toxicity was observed within 48 h of exposure. MTiO2 at concentrations of 25, 50 and 100 ppm was lethal for Artemia sp. with a significant difference (p ≤ .05) in relation to the control artificial sea water with LC50 value at 50 ppm. Analysis of optical and scanning electron microscopy revealed tissue damage and morphological changes in Artemia sp. nauplii instar II. By using confocal laser scanning microscopy, cell damage was observed due to the toxicity of MTiO2 at a concentration of 20, 50, and 100 ppm. The high mortality rate is related to the filtration of MTiO2 by Artemia sp. nauplii instar II due to the complete development of the digestive tract.

Effects of the Incorporation of Bioactive Particles on Physical Properties, Bioactivity and Penetration of Resin Enamel Infiltrant.

Purpose: The resinous infiltrant lacks remineralizing activity. This research aimed to develop and evaluate bioactivity, physico-mechanical properties and penetration of resin infiltrants containing Biosilicate or nanohydroxyapatite. Methods: Experimental resin infiltrant (ERI; 75/25 wt.% TEGDMA/BisEMA) was divided among the groups Pure Experimental (PE); ERI + Biosilicate 5 or 10% (Bio5; Bio10), ERI + 10% nanohydroxyapatite (Hap10), and Icon (DMG, Germany). Bioactivity was analyzed by SEM, EDS and FT-IR/ATR after soaking in SBF. Degree of conversion (DC), sorption and solubility (SO; SOL), flexural strength, modulus of elasticity (FS; E-modulus), contact angle (CA) and penetration were characterized. Extent of penetration was analyzed by treating white spot lesions (WSL) in human dental enamel samples with the infiltrants and subsequently analyzing specimens by confocal laser scanning microscopy. Data from each test were submitted to ANOVA and Tukey's tests (p < 0.01). Results: SEM, EDS and FT-IR showed the formation of precipitates and increase in the rates of Ca and P in the groups with bioactive particles, after storage in SBF. Hap10 showed higher DC and CA values than all the other groups. Groups Bio5 and Bio10 showed CA values similar to those of Icon, higher SO and SOL values, and reduction in other properties. All infiltrants were capable of penetrating into the WSLs. Conclusion: The incorporation of Biosilicate (5 or 10%) or nanohydroxyapatite (10%) into ERI induced mineral deposition on the surface and did not compromise infiltration and penetration into WSLs, however, compromising their physico-mechanical properties.

Curcumin-loaded microemulsion: formulation, characterization, and in vitro skin penetration.

OBJECTIVE: Formulation of curcumin in a microemulsion with a high loading capacity and that favors its penetration into the skin. SIGNIFICANCE: Take advantage of the properties of microemulsions to promote the penetration of curcumin into the skin, with the aim of enhancing its therapeutic effects. METHODS: Curcumin was formulated in microemulsions based on oleic acid (oil phase), Tween® 80 (surfactant), and Transcutol® HP (cosurfactant). The microemulsion formation area was mapped by constructing pseudo-ternary diagrams for surfactant:co-surfactant ratios 1:1, 1:2, and 2:1. Microemulsions were characterized through measurements of specific weight, refractive index, conductivity, viscosity, droplet size, and in vitro skin permeation studies. RESULTS: Nine microemulsions were prepared and characterized, showing clear, stable formulations with globule size dependent on the proportion of the components. The microemulsion with the highest loading capacity (60 mg/mL), based on Tween® 80, Transcutol® HP, oleic acid, and water (40:40:10:10) was able to penetrate the viable epidermis, finding a total amount of curcumin in the receptor medium at 24 h of 10.17 ± 9.7 µg/cm2. The distribution of curcumin in the skin, visualized by confocal laser scanning microscopy, showed that the maximum amount was located between 20 and 30 µm. CONCLUSION: The inclusion of curcumin in a microemulsion allows its passage into and through the skin. The localization of curcumin, especially in the viable epidermis, would be important for those cases where local conditions are sought to be treated.

Biofilm-forming microorganisms causing hospital-acquired infections from intravenous catheter: A systematic review.

The high prevalence of nosocomial infections is related to the use of medical insertion devices such as central venous catheters (CVCs). Most of the microorganisms causing nosocomial infections are biofilm producers, this characteristic allows them to adhere to abiotic surfaces and cause initial catheter infections that can lead to bloodstream infections. Our main goal in this systematic review was to evaluate the prevalence of biofilm among CVC-related infections, particularly among Intensive Care Unit (ICU) patients, in the studies applying different in vitro and in vivo methodologies. All studies reporting clinical isolates from patients with catheter-related nosocomial infections and biofilm evaluation published up to 24 June 2022 in the PubMed and Scopus databases were included. Twenty-five studies met the eligibility criteria and were included in this systematic review for analysis. Different methodologies were applied in the assessment of biofilm-forming microorganisms including in vitro assays, catheter-infected in vitro, and in vivo mouse models. The present study showed that between 59 and 100% of clinical isolates were able to form biofilms, and the prevalence rate of biofilm formation varied significantly between studies from different countries and regions. Among the clinical isolates collected in our study set, a wide variety of microorganisms including Gram-positive strains, Gram-negative strains, and Candida albicans were found. Many authors studied resistance mechanisms and genes related to biofilm development and surface adherence properties. In some cases, the studies also evaluated biofilm inhibition assays using various kinds of catheter coatings.

Effect of obturation technique on penetration of calcium silicate-based sealer into dentinal tubules after endodontic retreatment of mandibular premolars.

OBJECTIVE: Confocal laser scanning microscopy (CLSM) was used to investigate the penetration of endodontic sealers into the dentinal tubules after retreatment using two different obturation techniques. MATERIALS AND METHODS: Thirty mandibular premolars were prepared up to instrument F3 (ProTaper Universal, Dentsply) and filled with Endofill using the single cone technique. The canals were retreated using Mtwo instruments. Reobturation was performed with the Bio-C sealer mixed with a fluorophore dye (Fluo-3) using either the lateral condensation technique (group LC) or the single cone technique (group SC) (n = 15). Teeth were sectioned 2, 4, and 6 mm from the apex and analyzed with CLSM to assess the penetration of the sealer into the canal perimeter and the maximum depth of penetration of the sealer into the dentinal tubules. Data were analyzed using ANOVA and the Student-t and Holm-Sidak tests. RESULTS: In the apical segment, the penetrated perimeter was significantly higher in the LC group than in the SC group (p < 0.05); no significant difference was found in the middle and cervical segments (p > 0.05). In terms of penetration depth, no significant differences were found for any of the segments studied (p > 0.05). CONCLUSION: The LC technique promoted a higher percentage of canal circumference penetrated by the sealer than the SC technique in the apical segment after endodontic retreatment. CLINICAL RELEVANCE: CLSM demonstrated that the LC technique promoted a higher percentage of canal perimeter penetrated by the Bio-C sealer than the SC technique in the apical segment of mandibular premolars after retreatment.

Andean crops: kañiwa and tarwi flours used for the development of vegan gluten-free muffins.

BACKGROUND: The flours of two Andean crops, tarwi (Lupinus mutabilis) and kañiwa (Chenopodium pallidicaule Aellen), present an excellent nutritional profile for inclusion in vegan gluten-free muffin formulations. In this study, the proximal composition and techno-functional properties of tarwi and kañiwa flours, and the technological quality of batters and muffins (potato starch-based) formulated with 50% of these flours were evaluated. RESULTS: Andean flours have high protein, fiber, and fat content, and display high water and oil absorption. In premixes formulated with potato starch and Andean flours, a reduction in paste viscosity was observed due to starch dilution and lower water availability. Depending on their ability to interact with water, the batters formulated with these flours had a higher consistency. Confocal laser scanning micrographs showed that batters with Andean flours presented a complex matrix with dispersed starch granules surrounded by proteins and fiber fragments. Muffins made with Andean flours had a slightly lower specific volume than the control, but crumb hardness was not modified by tarwi flour (50%) and a mixture of tarwi (25%) and kañiwa (25%) flours. The intrinsic color of these flours modified crumb color, and their reducing sugar content also favored Maillard reactions in the crust. These color changes are desirable in gluten-free products because such products are frequently pale due to their high starch content. CONCLUSIONS: This study showed that tarwi and kañiwa flours are suitable for developing vegan, gluten-free muffins of good technological quality and improved nutritional profile, adding value to these underutilized ancestral flours. © 2022 Society of Chemical Industry.

Fluorescence Microscopy Methods for the Analysis and Characterization of Lignin.

Lignin is one of the most studied and analyzed materials due to its importance in cell structure and in lignocellulosic biomass. Because lignin exhibits autofluorescence, methods have been developed that allow it to be analyzed and characterized directly in plant tissue and in samples of lignocellulose fibers. Compared to destructive and costly analytical techniques, fluorescence microscopy presents suitable alternatives for the analysis of lignin autofluorescence. Therefore, this review article analyzes the different methods that exist and that have focused specifically on the study of lignin because with the revised methods, lignin is characterized efficiently and in a short time. The existing qualitative methods are Epifluorescence and Confocal Laser Scanning Microscopy; however, other semi-qualitative methods have been developed that allow fluorescence measurements and to quantify the differences in the structural composition of lignin. The methods are fluorescence lifetime spectroscopy, two-photon microscopy, Föster resonance energy transfer, fluorescence recovery after photobleaching, total internal reflection fluorescence, and stimulated emission depletion. With these methods, it is possible to analyze the transport and polymerization of lignin monomers, distribution of lignin of the syringyl or guaiacyl type in the tissues of various plant species, and changes in the degradation of wood by pulping and biopulping treatments as well as identify the purity of cellulose nanofibers though lignocellulosic biomass.

Penetrability of AH Plus Sealer into Dentinal Tubes after Retreatment Procedure with Rotary or Reciprocating Systems and Two Additional Cleaning Methods.

Introduction: The aim of this study was to evaluate the AH Plus sealer penetration into dentinal tubules after root canal retreatment followed by two additional cleaning methods. Methods and Materials: Thirty-two mandibular premolars with single canal were prepared up to the F4 ProTaper Universal instrument and filled by a single cone technique with the addition of Rhodamine B dye to Endofill sealer. For the retreatment procedure, the teeth were randomly divided into four experimental groups (n=8) as follows: Reciproc R40 with ultrasonic activation (RU), Reciproc R40 with sonic agitation (RS), ProTaper Next until X4 instrument with ultrasonic activation (PTNU), and ProTaper Next (X4) with sonic activation (PTNS). A new root canal filling was done using a System B technique, and the AH Plus sealer was mixed with Fluorescein dye. The roots were axial sectioned at 3, 5, and 7 mm from the apex and were assessed by a confocal laser scanning microscopy using the method of epifluorescence with wavelengths of absorption and emission for rhodamine B and fluorescein. In the obtained images, the sealer penetration into the dentinal tubules was evaluated. The data were converted into percentages and submitted to Mann-Whitney and Kruskal-Wallis followed by Dunn's tests (P<0.05). Results: In all groups were found penetration of the AH Plus into the dentinal tubules. Statistical difference was found (P<0.05) in the ProTaper Next groups in relation the Reciproc groups for 3 mm root canal level regardless of additional cleaning method used. For the other sections the sealer penetration was similar (P>0.05) for all groups. Conclusion: Based on this ex vivo study the retreatment with ProTaper Next showed significantly greater penetration of AH Plus sealer into dentinal tubules in the apical third. The additional cleaning methods did not improve the removal of filling material in all sections evaluated.

Antibacterial Efficacy of Triple Antibiotic Medication With Macrogol (3Mix-MP), Traditional Triple Antibiotic Paste, Calcium Hydroxide, and Ethanol Extract of Propolis: An Intratubular Dentin Ex Vivo Confocal Laser Scanning Microscopic Study.

INTRODUCTION: This study aimed to assess the effectiveness of antibacterial activity of medications used in regenerative endodontic treatment. METHODS: Sixty-seven dentin cylinders of single-rooted teeth were contaminated with a culture of Enterococcus faecalis (ATCC 29212; American Type Culture Collection, Manassas, VA) for 5 days. Samples were divided into 1 control group and the following experimental groups according to the medication applied: traditional triple antibiotic paste (TAP), clindamycin-modified TAP (mTAP), triple antibiotic medication with macrogol (3Mix-MP), clindamycin-modified 3Mix-MP (m3Mix-MP), calcium hydroxide (CH), and ethanol extract of propolis (EEP). After 14 days, the medications were removed, and the samples were submitted to confocal laser scanning microscopic analysis to quantify the percentage of viable bacteria. The distribution of data was confirmed by the Shapiro-Wilk test. The Kruskal-Wallis and Dunn tests were used for intergroup comparisons, and the Wilcoxon test was used for comparison between superficial and deep antibacterial efficacy for the same medication. The level of significance was set at P < .05. RESULTS: 3Mix-MP and m3Mix-MP presented significantly higher antibacterial efficacy compared with the other tested medications (P < .05), except for mTAP. mTAP was more effective than TAP (P < .05). The antibacterial efficacy of EEP and CH did not differ significantly from TAP and mTAP (P > .05). All medications showed effective antibacterial action compared with the control group (P < .05). CONCLUSIONS: 3Mix-MP and m3Mix-MP, which present extremely high concentrations of antibiotics (1500 mg/mL), were not more effective than mTAP at the concentration recommended by the American Association of Endodontists (5 mg/mL). Moreover, CH and EEP were as effective as TAP and mTAP.

Chemical and morphological changes of femtosecond laser-irradiated enamel using subablative parameters.

Chemical composition of dental enamel has a great relationship with the prevention of caries. The objective of the present work was to evaluate the chemical and morphological changes of femtosecond laser-irradiated enamel with subablative parameters using Raman spectroscopy, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM). Bovine incisor teeth were used to obtain 30 enamel specimens (5 × 5 mm2 ). The chemical composition of the control sample was analyzed by Raman spectrometry to acquire the absorption spectrum, delimiting the areas under the carbonate and phosphate bands. This analysis was used to evaluate the change in the chemical composition of the sample after irradiation. The specimens were irradiated (IRR) with a Ti:Sapphire laser system (pulsed and focused modes, femtosecond regime 70 fs, average power of 1 W and exposure time of 15 s). After irradiation, the areas under the carbonate and phosphate absorption bands were delimited in each specimen. Raman spectrometry data were analyzed using Student's t-test (α = 5%). By comparing the spectra of the IRR and non-irradiated (NI) specimens, the results showed a significant increase in the area value for the phosphate peaks and a significant reduction in the area value for the carbonate peak and the carbonate:phosphate ratio. CLSM and SEM analyses did not reveal structural alterations in the subsurface nor morphological alterations in the IRR enamel surface, respectively. It was concluded that femtosecond laser irradiation using subablative parameters reduced the carbonate content and the carbonate/phosphate ratio without altering the structure and morphology of the dental enamel.

Effect of Sodium Thiosulfate on Interfacial Adaptation and Penetration of an Epoxy Resin-Based Root Canal Sealer.

Introduction: Our study evaluated the impact of sodium thiosulfate (ST) irrigation, subsequent to sodium hypochlorite (NaOCl) and just before root canal filling, on the filling quality (interfacial adaptation and penetration segment) of an epoxy resin-based root canal sealer. Methods and Materials: Twenty single-rooted human teeth were prepared with the ProTaper system. The specimens were then divided into the following groups: 5.25% NaOCl irrigation (NaOCl group) and 5.25% NaOCl irrigation+0.5% sodium thiosulfate (NaOCl+ST group). The root canals were filled using single-cone technique with ProTaper F3 cones and AH-Plus sealer, labeled with rhodamine B dye to allow analysis under a confocal laser scanning microscopy (CLSM). All samples were sectioned at 2, 4, and 6 mm from the apex and prepared for CLSM analysis. The percentage of voids, gaps and dentinal sealer penetration segment of the canal were calculated at the apical, middle and coronal thirds. The non-parametric Mann-Whitney statistical test was used at 5% significance level. Results: Higher percentage of gaps and voids were observed at all root thirds of the NaOCl group when compared to the NaOCl+ST group (P<0.05). There was a significant increase in the penetration segment of NaOCl+ST group at the coronal and middle root third when compared to the NaOCl group (P<0.05). Conclusion: Our in vitro results showed that the use of ST as an antioxidant agent after NaOCl irrigation promoted a better interfacial adaptation and penetration of epoxy resin-based root canal fillings.

Chemical and mechanical influence of root canal irrigation on biofilm removal from lateral morphological features of simulated root canals, dentine discs and dentinal tubules.

AIM: To investigate the anti-biofilm efficacy of irrigation using a simulated root canal model, the chemical effect of irrigants against biofilms grown on dentine discs and their impact on biofilm viscoelasticity, the efficacy of the irrigants in decontaminating infected dentinal tubules and the capacity of bacteria to regrow. METHODOLOGY: Biofilm removal, viscoelastic analysis of remaining biofilms and bacterial viability were evaluated using a simulated root canal model with lateral morphological features, dentine discs and a dentinal tubule model, respectively. Experiments were conducted using a two-phase irrigation protocol. Phase 1: a modified salt solution (RISA) and sodium hypochlorite (NaOCl) were used at a low flow rate to evaluate the chemical action of the irrigants. Ultrasonic activation (US) of a chemically inert solution (buffer) was used to evaluate the mechanical efficacy of irrigation. Phase 2: a final irrigation with buffer at a high flow rate was performed for all groups. Optical coherence tomography (OCT), low load compression testing (LLCT) and confocal scanning laser microscopy analysis were used in the different models. One-way analysis of variance (anova) was performed for the OCT and LLCT analysis, whilst Kruskal-Wallis and Wilcoxon ranked tests for the dentinal tubule model. RESULTS: US and high flow rate removed significantly more biofilm from the artificial lateral canal. For biofilm removal from the artificial isthmus, no significant differences were found between the groups. Within-group analysis revealed significant differences between the steps of the experiment, with the exception of NaOCl. For the dentine discs, no significant differences regarding biofilm removal and viscoelasticity were detected. In the dentinal tubule model, NaOCl exhibited the greatest anti-biofilm efficacy. CONCLUSIONS: The mechanical effect of irrigation is important for biofilm removal. An extra high flow irrigation rate resulted in greater biofilm removal than US in the artificial isthmus. The mechanical effect of US seemed to be more effective when the surface contact biofilm-irrigant was small. After the irrigation procedures, the remaining biofilm could survive after a 5-day period. RISA and NaOCl seemed to alter post-treatment remaining biofilms.

A Novel Method Using Confocal Laser Scanning Microscopy for Three-Dimensional Analysis of Human Dental Enamel Subjected to Ceramic Bracket Debonding.

The aim of this in vitro study was to present a method using confocal laser scanning microscopy for three-dimensional analysis of human dental enamel subjected to ceramic bracket debonding. The labial enamel surfaces of three upper central incisors were prepared and mounted in the form of standardized specimens. A sample repositioning protocol was established to enable surface measurement and analysis before and after bracket debonding. Observations were made of representative areas measuring 1,280 × 1,280 µm2, in the center of the enamel samples, as well as of the total topography (2,500 × 3,500 µm) of the bonding areas provided by the equipment software. Noncontact three-dimensional high-resolution image analyses revealed the capabilities of the employed technique and methodology to permit the examination of specific characteristics and alterations on the surfaces, before and after the debonding and finishing procedures. The new protocol was effective to provide qualitative and quantitative assessments of changes on the same dental surfaces at different trial times. The methodology constitutes a feasible tool for revealing the effects of debonding of ceramic brackets on sound and previously injured dental enamel surfaces.

Complementary Dynamics of Banana Root Colonization by the Plant Growth-Promoting Rhizobacteria Bacillus amyloliquefaciens Bs006 and Pseudomonas palleroniana Ps006 at Spatial and Temporal Scales.

Banana (Musa acuminata) growth for commercial purposes requires high amounts of chemical fertilizers, generating high costs and deleterious effects on the environment. In a previous study, we demonstrated that two plant growth-promoting rhizobacteria (PGPR), Bacillus amyloliquefaciens Bs006 and Pseudomonas palleroniana Ps006, isolated in Colombia, could partially replace chemical fertilizers for banana seedling growth. In a second work, the effects of the two inoculants on banana transcripts were found to occur at different times, earlier for Bs006 and later for Ps006. This leads to the hypothesis that the two rhizobacteria have different colonization dynamics. Accordingly, the aim of this work was to analyze the dynamics of root colonization of the two PGPR, Bs006 and Ps006, on banana growth over a time frame of 30 days. We used fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy (CLSM), followed by three-dimensional reconstruction and quantitative image analysis. Bacillus amyloliquefaciens Bs006 abundantly colonized banana roots earlier (from 1 to 48 h), ectophytically on the rhizoplane, and then decreased. Pseudomonas palleroniana Ps006 was initially scarce, but after 96 h it increased dramatically and became clearly endophytic. Here we identify and discuss the potential genetic factors responsible for this complementary behavior. This information is crucial for optimizing the formulation of an effective biofertilizer for banana and its inoculation strategy.

Visualization of Arbuscular Mycorrhizal Fungal Extraradical Hyphae and Spores Vitality and Activity.

The hyphae and spores of arbuscular mycorrhizal (AM) fungi represent an essential component in the extraradical zone due to their role in nutrients and water uptake and as propagules that allow the perpetuation of the AM symbiosis over time, respectively. However, the attention of scientific literature is usually more focused on root colonization than on the study of the extraradical components of AM fungi, especially their vital, active, or functional fractions. This chapter presents some easy-to-use alternatives for staining vital, active, or functional structures of AM fungi for their subsequent microscopic visualization, such as the application of enzyme-based stains, NADPH formation, and also nucleus staining. Some modified methods for the extraction of mycelium from the soil are also presented.