Confocal Laser Scanning Microscopic Evaluation of Sealer Penetration in Root Canals of Teeth with the butterfly and Non-butterfly Effect: An In vitro Study

AIM: The study aimed to investigate the penetration depth of calcium hydroxide-based root canal sealer into buccolingual and mesiodistal aspects of roots with and without the butterfly effect at coronal and middle root sections. METHODS AND MATERIALS: Twenty single-rooted maxillary premolars were decoronated at the cementoenamel junction and viewed under a light microscope and grouped as Group 1 – butterfly (B) and Group 2 – non-butterfly according to the presence or absence of the effect. Canals were prepared till working length followed with copious irrigation. Canals were finally rinsed with 5 ml of 17% ethylenediaminetetraacetic acid solution and activated using EndoActivator followed by obturation using gutta-percha (warm vertical compaction technique) with Sealapex sealer. To provide fluorescence for confocal laser scanning microscopy (CLSM), the Sealapex was mixed with rhodamine B dye. Root sectioning yielded coronal and middle sections. CLSM was used to assess the penetration of the sealer. STATISTICAL ANALYSIS: Shapiro–Wilk test, unpaired “t-test.” RESULTS: Teeth with the butterfly effect had greater mean penetration buccolingually (905.2 μm) than mesiodistally (182.1 μm; p < 0.001). Coronal sections had greater penetration (517.4 μm) compared with the middle (354.6 μm). CONCLUSION: Sealapex sealer exhibited maximum tubular penetration in teeth with butterfly effect in buccolingual direction at the coronal third level.

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Identification TIMP-1 and TIMP-2 in Human Radicular Dentine - In Vitro Study

Journal of Academy of Dental Education ◽

10.18311/jade/2014/2390 ◽

2014 ◽

Vol 1 (2) ◽

pp. 12

Author(s):

Kandaswamy Eswar ◽

Rubin Mohamed Ismail ◽

Hannah Rosaline ◽

Nagendrababu Venkateshbabu ◽

Deivanayagam Kandaswamy

Keyword(s):

Confocal Laser Scanning Microscopy ◽

Laser Scanning ◽

Isotonic Saline ◽

In Vitro Study ◽

Laser Scanning Microscopy ◽

Confocal Laser Scanning ◽

Scanning Microscopy ◽

Confocal Laser ◽

Group 2

Aim: To identify TIMP – 1 and TIMP – 2 in human radicular dentine using confocal laser scanning microscopy. Materials and Methods: Thirty freshly extracted non carious human single rooted pre molars were obtained and stored in isotonic saline at -70°C prior to use. All the teeth were decoronated at the CEJ using a diamond. Teeth were divided into 2 groups (Group 1: TIMP-1 analysis n = 15; Group 2: TIMP-2 analysis n = 15). Teeth were sectioned using a hard tissue microtome, mounted and viewed under confocal laser scanning microscopy. Results: TIMP-1 and TIMP-2 were detected in radicular dentine and were seen to be distributed more towards the inner dentine layer closer to the pulp. Conclusion: Due to a shorter half life of TIMP-1 and 2 as compared to the MMP, there is a need to use MMP inhibitors prior to obturation of the root canal.

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Endotracheal tubes coated with a broad-spectrum antibacterial ceragenin reduce bacterial biofilm in an in vitro bench top model

Journal of Antimicrobial Chemotherapy ◽

10.1093/jac/dkab019 ◽

2021 ◽

Author(s):

María Consuelo Latorre ◽

María Jesús Pérez-Granda ◽

Paul B Savage ◽

Beatriz Alonso ◽

Pablo Martín-Rabadán ◽

...

Keyword(s):

Biofilm Formation ◽

Laser Scanning ◽

In Vitro Study ◽

Bacterial Biofilm ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Endotracheal Tubes ◽

Clinical Strains ◽

Number Of Cells

Abstract Background Ventilator-associated pneumonia is one of the most common nosocomial infections, caused mainly by bacterial/fungal biofilm. Therefore, it is necessary to develop preventive strategies to avoid biofilm formation based on new compounds. Objectives We performed an in vitro study to compare the efficacy of endotracheal tubes (ETTs) coated with the ceragenin CSA-131 and that of uncoated ETTs against the biofilm of clinical strains of Pseudomonas aeruginosa (PA), Escherichia coli (EC) and Staphylococcus aureus (SA). Methods We applied an in vitro bench top model using coated and uncoated ETTs that were treated with three different clinical strains of PA, EC and SA for 5 days. After exposure to biofilm, ETTs were analysed for cfu count by culture of sonicate and total number of cells by confocal laser scanning microscopy. Results The median (IQR) cfu/mL counts of PA, EC and SA in coated and uncoated ETTs were, respectively, as follows: 1.00 × 101 (0.0–3.3 × 102) versus 3.32 × 109 (6.6 × 108–3.8 × 109), P < 0.001; 0.0 (0.0–5.4 × 103) versus 1.32 × 106 (2.3 × 103–5.0 × 107), P < 0.001; and 8.1 × 105 (8.5 × 101–1.4 × 109) versus 2.7 × 108 (8.6 × 106–1.6 × 1011), P = 0.058. The median (IQR) total number of cells of PA, EC and SA in coated and non-coated ETTs were, respectively, as follows: 11.0 [5.5–not applicable (NA)] versus 87.9 (60.5–NA), P = 0.05; 9.1 (6.7–NA) versus 62.6 (42.0–NA), P = 0.05; and 97.7 (94.6–NA) versus 187.3 (43.9–NA), P = 0.827. Conclusions We demonstrated significantly reduced biofilm formation in coated ETTs. However, the difference for SA was not statistically significant. Future clinical studies are needed to support our findings.

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Consequences of topoisomerase II inhibition in early embryogenesis of Drosophila revealed by in vivo confocal laser scanning microscopy

Journal of Cell Science ◽

10.1242/jcs.104.4.1175 ◽

1993 ◽

Vol 104 (4) ◽

pp. 1175-1185 ◽

Cited By ~ 12

Author(s):

P. Buchenau ◽

H. Saumweber ◽

D.J. Arndt-Jovin

Keyword(s):

Confocal Laser Scanning Microscopy ◽

Laser Scanning ◽

Topoisomerase Ii ◽

Metaphase Plate ◽

Laser Scanning Microscopy ◽

Confocal Laser Scanning ◽

Scanning Microscopy ◽

Confocal Laser

The regulation of DNA topology by topoisomerase II from Drosophila melanogaster has been studied extensively by biochemical methods but little is known about its roles in vivo. We have performed experiments on the inhibition of topoisomerase II in living Drosophila blastoderm embryos. We show that the enzymatic activity can be specifically disrupted by microinjection of antitopoisomerase II antibodies as well as the epipodophyllotoxin VM26, a known inhibitor of topoisomerase II in vitro. By labeling the chromatin of live embryos with tetramethylrhodamine-coupled histones, the effects of inhibition on nuclear morphology and behaviour was followed in vivo using confocal laser scanning microscopy. Both the antibodies and the drug prevented or hindered the segregation of chromatin daughter sets at the anaphase stage of mitosis. In addition, high concentrations of inhibitor interfered with the condensation of chromatin and its proper arrangement into the metaphase plate. The observed effects yielded non-functional nuclei, which were drawn into the inner yolk mass of the embryo. Concurrently, undamaged nuclei surrounding the affected region underwent compensatory division, leading to the restoration of the nuclear population, and thereby demonstrating the regulative capacity of Drosophila blastoderm embryos.

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Percentage of Gutta-Percha-, Sealer-, and Void-Filled Areas in Oval-Shaped Root Canals Obturated with Different Filling Techniques: A Confocal Laser Scanning Microscopy Study

European Journal of Dentistry ◽

10.1055/s-0040-1701543 ◽

2020 ◽

Vol 14 (01) ◽

pp. 008-012

Author(s):

Vinicio Hidemitsu Goto Hirai ◽

Ricardo Machado ◽

Maria Carolina Lucato Budziak ◽

Lucila Piasecki ◽

Alexandre Kowalczuck ◽

...

Keyword(s):

Confocal Laser Scanning Microscopy ◽

Laser Scanning ◽

Continuous Wave ◽

Laser Scanning Microscopy ◽

Confocal Laser Scanning ◽

Scanning Microscopy ◽

Confocal Laser ◽

Root Canals ◽

Gutta Percha

Abstract Objective This study compared different obturation techniques, analyzing percentage of areas filled with gutta-percha, sealer, and voids (PGFA, PSFA, and PVFA, respectively) in oval-shaped root canals. Materials and Methods A total of 60 extracted human mandibular central incisors were decoronated, instrumented, and irrigated using the same protocol. After drying, the root canal was filled with AH Plus labeled with 0.1% rhodamine B dye using a Lentulo spiral. The filling procedure was performed by dividing the teeth into four groups according to the respective technique: G1, cold lateral condensation; G2, continuous wave of condensation; G3, modified cold lateral condensation using an F3 master cone; and G4, modified continuous wave of condensation using an ISO (International Organization for Standardization) sized 30 gutta-percha cone. Then, slices measuring 1.5 mm in thickness were obtained 3 and 6 mm from the apex and evaluated by confocal laser scanning microscopy to determine PGFA, PSFA, and PVFA. Statistical Analysis The data were analyzed statistically with analysis of variance and Games-Howell’s tests (p = 0.05). Results The groups showed no significant differences in the apical third (3 mm from the apex). In the middle third (6 mm from the apex), G3 and G1 showed higher PGFA and PVFA, respectively. G3 showed lower PSFA than G2 and G4. Both cold techniques (G1 and G3) promoted lower PSFA than both warm techniques (G2 and G4). Conclusions Notwithstanding the limitations of this in vitro study, PGFA, PSFA, and PVFA ranged significantly only in the middle third, as observed by the different filling techniques. Higher PGFA and PVFA values were obtained for G3 and G1, respectively. Both cold techniques promoted lower PSFA than both warm techniques.

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