Antimicrobial efficacy and biocompatibility of extracts from Cryptocarya species

This study evaluated the efficacy of Cryptocarya spp extracts on biofilm of Candida albicans and its biocompatibility. Mature biofilm of C. albicans was formed on denture base acrylic resin samples and the fungicidal effect of the extracts was evaluated by Alamar Blue® assay, counting colony-forming units (CFU/mL) and confocal laser scanning microscopy (CLSM). Cytotoxicity of extracts from Cryptocarya species was evaluated by AlamarBlue® assay, using normal oral keratinocytes (NOK) cells. In additional, Analysis of plant extracts by ultra-high-performance liquid chromatography–diode array detector–tandem mass spectrometry (UPLC-DAD-MS) was performed. The results showed significant reduction in the cellular metabolism and in the number of CFU/mL of C. albicans (p<0.05). The concentration of 0.045 g/mL completely inhibited the number of CFU/mL. Regarding cytotoxicity, all extracts decreased cell viability compared to the control group. CLSM analysis showed predominance of live cells, but with a great difference between the groups. Antimicrobial activity of extracts from Cryptocarya on C. albicans biofilm was confirmed. However, all extracts showed toxicity on NOK cells.

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Effect of reciprocating instrumentation on chlorhexidine substantivity on human dentin: chemical analysis followed by confocal laser microscopy

Bioscience Journal ◽

10.14393/bj-v37n0a2021-54518 ◽

2021 ◽

Vol 37 ◽

pp. e37038

Author(s):

Matheus Albino Souza ◽

Fernanda Duda Bonatto ◽

Afonso Cristiano Fleck da Silva ◽

Ezequiel Santin Gabrielli ◽

Felipe Trentin Motter ◽

...

Keyword(s):

High Performance ◽

Laser Scanning ◽

Ex Vivo ◽

Experimental Tests ◽

Laser Scanning Microscopy ◽

Live Cells ◽

Confocal Laser ◽

Water Control ◽

Human Teeth ◽

Root Canals

The present research analyzed the reciprocating instrumentation associated to chlorhexidine (CHX) substantivity as its correlation with E. faecalis viability in ex vivo root canals. Eighty extracted single-rooted human teeth were used, being 40 to high-performance liquid chromatography (HPLC) and 40 to confocal laser scanning microscopy (CLSM). In both, teeth were decoronated and the cervical third was prepared. In the CLSM analysis, the root canals were inoculated with E. faecalis for 14 days. Samples were divided into 4 groups (n=10) according to instrumentation technique: no instrumentation and irrigation with distilled water (control); manual instrumentation (K-File); rotary instrumentation (ProTaper Next); and reciprocating instrumentation (Reciproc R25). Two percent chlorhexidine was applied as irrigating substance in experimental groups. Longitudinal grooves resulted in 2 halves root and 20 proof bodies in each group. Samples were divided by chance in two groups (n=10) and the outcomes were evaluated after two days and one week. The retained chlorhexidine and live cells after instrumentation techniques in each evaluation time was measured by HPLC and CLSM, respectively. Specific analysis was applied for experimental tests (p≤0.05). Both rotary as well as reciprocating techniques significantly reduced the amount of chlorhexidine on dentin in all observation periods (p<0.05). After evaluation times, all experimental groups presented lower live cells compared to control, but without statistically difference. Intragroup comparisons in times of evaluation showed no differences in instrumentation techniques, in chlorhexidine retention and number of live cells (p>0.05). Reciprocating instrumentation does not interfere on chlorhexidine substantivity.

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Repurposing Napabucasin as an Antimicrobial Agent against Oral Streptococcal Biofilms

BioMed Research International ◽

10.1155/2020/8379526 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Xinyi Kuang ◽

Tao Yang ◽

Chenzi Zhang ◽

Xian Peng ◽

Yuan Ju ◽

...

Keyword(s):

Dental Caries ◽

Laser Scanning ◽

Antibacterial Activities ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Oral Streptococci ◽

Oral Keratinocytes ◽

Microbial Composition ◽

Biofilm Inhibition ◽

Multispecies Biofilms

Objectives. Disruption of microbial biofilms is an effective way to control dental caries. Drug resistance and side effects of the existing antimicrobials necessitate the development of novel antibacterial agents. The current study was aimed at investigating the antibacterial activities of the repurposed natural compound napabucasin against oral streptococci. Methods. The minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibition concentration, and minimum biofilm reduction concentration of Streptococcus mutans, Streptococcus gordonii, and Streptococcus sanguinis were examined by a microdilution method. Cytotoxicity of napabucasin against human oral keratinocytes, human gingival epithelia, and macrophage RAW264.7 was evaluated by CCK8 assays. The dead/live bacterium and exopolysaccharide in the napabucasin-treated multispecies biofilms were evaluated by confocal laser scanning microscopy. Microbial composition within the napabucasin-treated biofilms was further visualized by fluorescent in situ hybridization and qPCR. And the cariogenicity of napabucasin-treated biofilms was evaluated by transverse microradiography. Results. Napabucasin exhibited good antimicrobial activity against oral streptococcal planktonic cultures and biofilms but with lessened cytotoxicity as compared to chlorhexidine. Napabucasin reduced the cariogenic S. mutans and increased the proportion of the commensal S. gordonii in the multispecies biofilms. More importantly, napabucasin significantly reduced the demineralization capability of biofilms on tooth enamels. Conclusion. Napabucasin shows lessened cytotoxicity and comparable antimicrobial effects to chlorhexidine. Repurposing napabucasin may represent a promising adjuvant for the management of dental caries.

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Visualizing NO in live cells by confocal laser scanning microscopy

Chinese Optics Letters ◽

10.3788/col201210.s11701 ◽

2012 ◽

Vol 10 (s1) ◽

pp. S11701-311703 ◽

Cited By ~ 2

Author(s):

Yimei Huang Yimei Huang ◽

Hongqin Yang Hongqin Yang ◽

Xiuqiu Shen Xiuqiu Shen ◽

Yuhua Wang Yuhua Wang ◽

Liqin Zheng Liqin Zheng ◽

...

Keyword(s):

Confocal Laser Scanning Microscopy ◽

Laser Scanning ◽

Laser Scanning Microscopy ◽

Confocal Laser Scanning ◽

Scanning Microscopy ◽

Live Cells ◽

Confocal Laser

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Susceptibility of Mature Staphylococcus Biofilms to Chinese Herbal Decoction Sanhuang Jiedu: An In Vitro Study

BioMed Research International ◽

10.1155/2020/7473942 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

Shaoe Zhang ◽

Xiao Wang ◽

Xiaotao Shi ◽

Honglue Tan ◽

Himanshu Garg

Keyword(s):

Laser Scanning ◽

Multidrug Resistant ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Control Group ◽

Antibiofilm Activity ◽

Dependent Manner ◽

Chinese Herbal ◽

Titanium Surfaces

Background. External socking and washing with the Chinese herbal Sanhuang Jiedu decoction (SHJD) can effectively control local limb infections with bone and implant exposure. However, the antibiofilm activities of this decoction in vitro have not yet been investigated. Therefore, the aim of this study was to examine the effects and characteristics of SHJD on the mature biofilms of multidrug-resistant staphylococci on a titanium surface. Methods. Biofilm-forming methicillin-resistant Staphylococcus epidermidis ATCC 35984 and S. aureus ATCC 43330, and non-biofilm-forming S. epidermidis ATCC 12228 were selected as the experimental strains. The mature biofilms were prepared on titanium surfaces. The five experimental groups were based on dilution concentrations (DC) of SHJD: the control group (biofilm incubated with 0.85% NaCl solution), the SHJD (DC:1/8) group (initial SHJD solution was diluted 1/8), the SHJD (DC:1/4) group, the SHJD (DC:1/2) group, and the SHJD (DC:1/1) group (initial SHJD solution). The effects of SHJD on the mature biofilms were observed with the bacterial spread plate method, crystal violet (CV) staining, scanning electron microscopy, and confocal laser scanning microscopy. Results. After culture in tryptic soy broth for 72 h, ATCC 43300 and ATCC 35984 produced mature biofilms and ATCC 12228 did not. The optical density value of ATCC 12228 was 0.11 ± 0.02 , significantly lower than that of ATCC 35984 ( 0.42 ± 0.05 ) or ATCC 43300 ( 0.41 ± 0.03 ) ( P < 0.05 ). The mature biofilms of ATCC 43300 and ATCC 35984 clearly disintegrated when incubated for 12–24 h with SHJD (DC:1/1) or SHJD (DC:1/2), showing only scattered bacterial adhesion. In the SHJD (DC:1/4) group, although many residual bacterial colonies still clustered together, presenting a biofilm structure, it was very looser than that in the SHJD (DC:1/8) group in which the biofilm was similar to that in the control group. For ATCC 12228, only colony adhesion was observed, and the number of colonies decreased as the concentration of SHJD or the culture period increased. The quantitative results for the bacterial spread plate and CV staining showed significant differences between the SHJD groups ( P < 0.05 ). Conclusion. SHJD has antibiofilm activity against multidrug-resistant Staphylococcus strains. It weakens or disrupts already-formed mature biofilms on titanium surfaces in a concentration- and incubation time-dependent manner.

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Analysis of Shear Bond Strength and Morphology of Er:YAG Laser-Recycled Ceramic Orthodontic Brackets

BioMed Research International ◽

10.1155/2016/7276287 ◽

2016 ◽

Vol 2016 ◽

pp. 1-6 ◽

Cited By ~ 1

Author(s):

Ruo-qiao Han ◽

Kai Yang ◽

Ling-fei Ji ◽

Chen Ling

Keyword(s):

Bond Strength ◽

Shear Bond Strength ◽

Laser Scanning ◽

Er:Yag Laser ◽

Statistical Significance ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Control Group ◽

Test Machine ◽

Ceramic Brackets

Objective. The aim of this study was to compare the recycling of deboned ceramic brackets via an Er:YAG laser or via the traditional chairside processing methods of flaming and sandblasting; shear bond strength and morphological changes were evaluated in recycled brackets versus new brackets.Materials and Methods. 3M Clarity Self-Ligating Ceramic Brackets with a microcrystalline base were divided into groups subjected to flaming, sandblasting, or exposure to an Er:YAG laser. New ceramic brackets served as a control group. Shear bond strengths were determined with an Electroforce test machine and tested for statistical significance through analysis of variance. Morphological examinations of the recycled ceramic bracket bases were conducted with scanning electron microscopy and confocal laser scanning microscopy. Residue on the bracket base was analyzed with Raman spectroscopy.Results. Faded, dark adhesive was left on recycled bracket bases processed via flaming. Adhesive was thoroughly removed by both sandblasting and exposure to an Er:YAG laser. Compared with new brackets, shear bond strength was lower after sandblasting (p<0.05), but not after exposure to an Er:YAG laser. The Er:YAG laser caused no damage to the bracket.Conclusion. Er:YAG lasers effectively remove adhesive from the bases of ceramic brackets without damaging them; thus, this method may be preferred over other recycling methods.

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Polyamines Are Essential for the Formation of Plague Biofilm

Journal of Bacteriology ◽

10.1128/jb.188.7.2355-2363.2006 ◽

2006 ◽

Vol 188 (7) ◽

pp. 2355-2363 ◽

Cited By ~ 121

Author(s):

Chandra N. Patel ◽

Brian W. Wortham ◽

J. Louise Lines ◽

Jacqueline D. Fetherston ◽

Robert D. Perry ◽

...

Keyword(s):

High Performance ◽

Laser Scanning ◽

Double Mutant ◽

Arginine Decarboxylase ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Crystal Violet Staining ◽

Liquid Chromatography Mass Spectrometry ◽

Double Deletion ◽

Single Deletion

ABSTRACT We provide the first evidence for a link between polyamines and biofilm levels in Yersinia pestis, the causative agent of plague. Polyamine-deficient mutants of Y. pestis were generated with a single deletion in speA or speC and a double deletion mutant. The genes speA and speC code for the biosynthetic enzymes arginine decarboxylase and ornithine decarboxylase, respectively. The level of the polyamine putrescine compared to the parental speA + speC + strain (KIM6+) was depleted progressively, with the highest levels found in the Y. pestis ΔspeC mutant (55% reduction), followed by the ΔspeA mutant (95% reduction) and the ΔspeA ΔspeC mutant (>99% reduction). Spermidine, on the other hand, remained constant in the single mutants but was undetected in the double mutant. The growth rates of mutants with single deletions were not altered, while the ΔspeA ΔspeC mutant grew at 65% of the exponential growth rate of the speA + speC + strain. Biofilm levels were assayed by three independent measures: Congo red binding, crystal violet staining, and confocal laser scanning microscopy. The level of biofilm correlated to the level of putrescine as measured by high-performance liquid chromatography-mass spectrometry and as observed in a chemical complementation curve. Complementation of the ΔspeA ΔspeC mutant with speA showed nearly full recovery of biofilm to levels observed in the speA + speC + strain. Chemical complementation of the double mutant and recovery of the biofilm defect were only observed with the polyamine putrescine.

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Effect of Calcium-phosphate Desensitizers on Staining Susceptibility of Acid-eroded Enamel

Operative Dentistry ◽

10.2341/18-024-l ◽

2019 ◽

Vol 44 (3) ◽

pp. 281-288 ◽

Cited By ~ 2

Author(s):

KY Kyaw ◽

M Otsuki ◽

MS Segarra ◽

N Hiraishi ◽

J Tagami

Keyword(s):

Surface Roughness ◽

Calcium Phosphate ◽

Repeated Measures ◽

Laser Scanning ◽

Artificial Saliva ◽

Color Difference ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Control Group ◽

Before And After

SUMMARY Objective: To investigate the effect of calcium-phosphate–based desensitizers, Teethmate AP paste (TMAP) and Teethmate Desensitizer (TMD) (Kuraray Noritake Dental, Tokyo, Japan), on the prevention of staining on acid-eroded enamel. Methods and Materials: Forty polished enamel samples (4×4×1 mm) from bovine incisors were randomly divided into five groups (n=8). After immersion in 50 mL of 0.5% citric acid (pH 2.5) for 15 minutes to form acid-eroded surfaces, the surfaces were subjected to different treatments with TMAP, TMD, and NaF (0.21% means 950 ppm) for five minutes. Another eroded group was not treated with desensitizer. For the control group, the samples were not eroded or treated. All the samples were stored in artificial saliva (AS) at pH 7.2 for 24 hours at 37°C. The TMAP, TMD, or NaF was reapplied at eight and 16 hours during the 24 hours of storage time. The surface roughness (Sa) was evaluated following ISO 25178 for surface texture using confocal laser scanning microscopy (VK-X 150 series, Keyence, Osaka, Japan) before acid erosion, after acid erosion, and after 24 hours of incubation in AS. Afterward, the color difference was measured with a dental colorimeter (Shade Eye NCC, Shofu, Kyoto, Japan) before and after staining with tea solution. Results: One-way repeated measures analysis of variance showed that acid erosion significantly increased Sa (p<0.001). TMAP- and TMD-treated groups exhibited lower Sa values than the NaF group and the no-desensitizer treatment group. The greatest staining was observed in the NaF group and the no-desensitizer group, while the TMAP and TMD groups significantly decreased the formation of stains. Conclusions: Acid-eroded enamel increased surface roughness and tended to absorb more stains. However, the application of TMAP and TMD moderated the roughness and thus prevented the formation of extrinsic stains.

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131 APOPTOSIS AND DEVELOPMENT OF IN VITRO-PRODUCED BOVINE EMBRYOS EXPOSED TO SUPRAPHYSIOLOGICAL CONCENTRATIONS OF INSULIN-LIKE GROWTH FACTOR-1 (IGF-1)

Reproduction Fertility and Development ◽

10.1071/rdv21n1ab131 ◽

2009 ◽

Vol 21 (1) ◽

pp. 165

Author(s):

M. A. Velazquez ◽

H. Niemann

Keyword(s):

Laser Scanning ◽

Apoptotic Cell ◽

Polycystic Ovary ◽

Statistical Significance ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Control Group ◽

Fisher Exact Test ◽

Bovine Embryos

It has been hypothesized that high non-physiological IGF-1 levels are partially responsible for the recurrent pregnancy loss observed in women with the polycystic ovary syndrome (Eng GS et al. 2007 Diabetes 56, 2228–2234). The aim of this study was to determine the effect of supraphysiological concentrations of IGF-1 on blastocyst production and the occurrence of apoptosis in bovine embryos, which are a good model for human embryo development (Baumann CG et al. 2007 Mol. Reprod. Dev. 74, 1345–1353). COC obtained by slicing from abattoir ovaries were matured (TCM-199, Sigma) for 24 h and fertilized (Fert-TALP) for 18 h (Day 0) in vitro. Two different IGF-1 (Recombinant human IGF-1, R&D Systems GmbH, Wiesbaden, Germany) concentrations (supraphysiological = 1000 ng mL–1 and physiological = 100 ng mL–1) were added to the culture media (Synthetic oviduct fluid/BSA) and compared with a control group (no IGF-1 supplementation). On Day 8, blastocyst rates (22 replicates) were recorded and DNA degradation was detected in blastocyst nuclei using a cell death detection kit (Roche Diagnostics GmbH, Mannheim, Germany) based on the terminal deoxinucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) principle. Embryos (n = 27 [control], n = 29 [both IGF-1 groups]) from 4 replicates were examined by confocal laser scanning microscopy. Data were analyzed by ANOVA and the Fisher exact test using the SigmaStat 2.0 software package (Jandel Scientific, San Rafael, CA). Cleavage was numerically improved by both, 1000 (59.1 ± 1.8) and 100 (58.2 ± 2.8) ng IGF-1 over controls (53.5 ± 2.2), but the differences did not reach statistical significance (P = 0.22). The proportion of hatched blastocysts was enhanced by 100 (5.8 ± 1.0, P = 0.03) and 1000 (5.1 ± 0.7, P = 0.03) ng IGF-1 compared to controls (2.8 ± 0.6). Total blastocyst rate was increased by 100 ng IGF-1 (34.4 ± 1.9, P = 0.02) over controls (28.3 ± 1.7), but not by 1000 ng IGF-1 (29.1 ± 1.6 P = 0.75). The 100 ng IGF-1 group (38.5 ± 3.7) had fewer degenerated embryos (P = 0.01) compared to 1000 ng IGF-1 (49.7 ± 3.3). The proportion of embryos displaying at least one apoptotic cell was greater in the 1000 ng IGF-1 group over controls (96% v. 77% P = 0.04). The number of blastomeres with TUNEL-positive nuclei per embryo was higher in the supraphysiological group (5.5 ± 0.6, P < 0.001) compared with the control (2.3 ± 0.4) and the physiological group (2.5 ± 0.3). There were no significant differences between the control and the 100 ng IGF-1 group in this regard (P = 0.49). In conclusion, supraphysiological concentrations of IGF-1 do not increase blastocyst production but increase levels of apoptosis in bovine embryos produced in vitro. M. A. V. is in the PhD program of the University of Veterinary medicine, Hannover, Germany, and is supported by the German Academic Exchange Service (DAAD)

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The Effect of A2E On The Levels of ROS and The Distribution of A2E in RPE Cells Exposed to Blue Light

10.21203/rs.3.rs-1098430/v1 ◽

2021 ◽

Author(s):

Maomei Luo ◽

Chun Zeng ◽

Shu Wang ◽

Shanjun Cai

Keyword(s):

Blue Light ◽

High Performance ◽

Laser Scanning ◽

Light Exposure ◽

Damage Model ◽

Confocal Laser ◽

Control Group ◽

Rpe Cells ◽

Light Group

Abstract AimsTo establish the N-retinylidene-N-retinylethanolamine(A2E) and blue light induced RPE cells damage model to explore the regularity of distribution of A2E and the levels of reactive oxygen species(ROS).MethodsThe fourth to sixth generation of human RPE cells in vitro were divided into five groups randomly: control group, blue light group, A2E-loaded group, A2E-loaed+blue light group and A2E-loaded+blue light +nifedipine group. The levels of ROS in cytoplasm by DCFH-DA staining was assayed by flow cytometry. The concentration of A2E in cytoplasm and lysosomes were assayed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS). The fluorescence intensity of A2E in lysosomes by Lysotracker redDND-99 staining was assayed by confocal laser scanning microscope. ResultsExposure to blue light and/or A2E could increase the levels of ROS in RPE cells, and nifedipine could inhibit oxidative stress response and reduce ROS levels. By HPLC-MS, it was found that A2E was not detected in the groups without load A2E, and A2E levels in cytoplasm and lysosomes decreased after light exposure. The green fluorescence produced by A2E loaded on RPE cells was mostly coincident with the red fluorescence labeled by lysosomes.ConclusionBlue light and A2E can increase the ROS levels of RPE cells and both have a synergistic effect. A2E is mainly concentrated in lysosomes, which is reduced by oxidation under blue light irradiation, damages lysosomal membrane with oxidized species of A2E, and leaks out from lysosomes.

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Effects of cigarette smoking on the growth of Streptococcus mutans biofilms: an in vitro study

10.21203/rs.3.rs-221658/v1 ◽

2021 ◽

Author(s):

Ye Han

Keyword(s):

Treatment Group ◽

Cigarette Smoking ◽

Streptococcus Mutans ◽

Laser Scanning ◽

Water Soluble ◽

Laser Scanning Microscopy ◽

Extracellular Polysaccharides ◽

Confocal Laser ◽

Control Group

Abstract This study aimed to investigate the differences in growth and virulence (EPSs and acidogenicity) of Streptococcus mutans biofilms (S. mutans) according to the different times of cigarette smoking (CS) treatment. S. mutans biofilms (74-hour-old) were formed on saliva-coated hydroxyapatite disks. The biofilms were treated with CS at different times per day (one time, three times, and six times/day). The control group did not receive CS treatment. Acidogenicity, dry weight, colony-forming units, water-soluble/insoluble extracellular polysaccharides, and intracellular polysaccharides were analyzed and confocal laser scanning microscopy images were obtained of the 74-h-old biofilms. The 74-h-old biofilms on sHA discs in the 6 times/day CS treatment group showed the lowest biofilm accumulation and extracellular polysaccharide amount compared with the control group and other CS treatment groups. In the CLSM study, the biofilms in the six times/day CS treatment group also showed the lowest bacterial count (live and dead cells) and EPS biovolume. CS has an obvious inhibition on the growth of S. mutans biofilms, the degree of inhibition is proportional to the number of CS treatments.

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