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

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.

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

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.

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

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)

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

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.

scholarly journals Curcumin induces apoptosis in trophoblast model cell line

2018 ◽  
Vol 27 (2) ◽  
Author(s):  
Tatit Nurseta ◽  
Yahya Irwanto ◽  
I W.A. Wiyasa ◽  
Rahajeng Rahajeng ◽  
Imelda Imelda ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Nuclear Translocation ◽  
Hydatidiform Mole ◽  
In Vitro Study ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Control Group ◽  
Trophoblast Cells ◽  
Enos Expression

Background: Several studies have reported that curcumin exerts chemopreventive effects in various type of cancers, through several mechanisms, however, the effect of curcumin on carcinogenesis in patients with hydatidiform mole has not yet been investigated. This study was conducted to evaluate the effect of curcumin on apoptosis, proliferation, and nuclear translocation of endothelial nitricoxide synthase in trophoblast cells induced by estradiol in complete hydatidiform mole (CHM).Methods: In this in vitro study, trophoblast cells were divided into six groups, the control group (trophoblast cells were exposed to 100 pg/mL of 17-β estradiol) and the treatment group (trophoblast cells were exposed to 100 pg/mL of 17-β estradiol in the presence of curcumin with doses: 50, 100, 200, 400, and 800 µM). At the end of study, the cell proliferation was analyzed using MTT assay and apoptosis with TUNEL test in each group thropoblast cell. eNOS translocation was assayed using confocal laser scanning microscopy at the various dose of curcumin.Results: Curcumin at the doses of 200, 400, and 800 µM significantly decreased the proliferation and increased the apoptotic index in curcumin-treated group compared to those in the control group (p<0.05). All doses of curcumin treatment significantly decreased the nuclear eNOS expression compared to that in the control group. The three highest doses of curcumin increased cytoplasmic eNOS expression compared to that in control group.Conclusion: Curcumin inhibits the proliferation and modulates the apoptosis of trophoblast cells induced by estradiol in CHM involvement.

scholarly journals An In Vitro Comparative Evaluation of the Effects of Different Bleaching Agents on Marginal Integrity of Two Different Restorative Resin Materials: An Analysis by Confocal Laser Microscopy

2019 ◽  
Vol 07 (03) ◽  
pp. 114-122
Author(s):  
Nitesh Goyal ◽  
Yogesh Kumar ◽  
Neetu Jindal ◽  
Renu Aggarwal
Keyword(s):  
Laser Scanning ◽  
Composite Resin ◽  
Root Surface ◽  
Composite Resins ◽  
Tooth Bleaching ◽  
Confocal Laser ◽  
Control Group ◽  
Class V ◽  
Nanohybrid Composite

Abstract Introduction In our society, with the advances in cosmetic consciousness, well-aligned, properly contoured, white and clean teeth are a symbol of health and standard of beauty. Thus, tooth bleaching has become popular cosmetic therapy among patients and dentists. Microleakage is a major challenge to the success rate for all type of restorations. Materials and Methods Eighty maxillary human molars were collected and evaluated and Class V cavities 5×3×2 mm size were prepared with straight carbide bur (SS White) on the buccal and palatal surfaces at the cementoenamel junction of all the samples, so that the upper margin would be in enamel and lower margins on root surface. In total, 160 class V cavities were prepared on maxillary molars. Then samples were randomly divided into 3 groups (microfilled and nanohybrid composite resins and control group) and 3 subgroups (Pola Office, Whiteness HP Blue, Whiteness Perfect) according to the type of composite resins and bleaching agents used. After application of bleaching agents, all samples were immersed in a contrast solution of rhodamine B fluorescent dye for 24 hours. Then using a diamond disc samples were sectioned buccolingually through the center of the restorations and sectioned teeth were evaluated for marginal microleakage under confocal laser scanning microscope to check the extent of dye penetration. Results When all composite resin groups were compared with all the bleaching agents, at occlusal and gingival levels, lowest marginal leakage scores were obtained with the microfilled composite resin bleached with Whiteness HP blue, and in nanohybrid composite lowest microleakage score were obtained with Pola office bleach at both levels.

scholarly journals Antimicrobial efficacy and biocompatibility of extracts from Cryptocarya species

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0261884
Author(s):  
Jacqueline de Oliveira Zoccolotti ◽  
Alberto José Cavalheiro ◽  
Camilla Olga Tasso ◽  
Beatriz Ribeiro Ribas ◽  
Túlio Morandin Ferrisse ◽  
...  
Keyword(s):  
High Performance ◽  
Laser Scanning ◽  
Acrylic Resin ◽  
Laser Scanning Microscopy ◽  
Live Cells ◽  
Confocal Laser ◽  
Array Detector ◽  
Control Group ◽  
Oral Keratinocytes ◽  
Denture Base

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.

scholarly journals Reduction of an in vitro Intraradicular Multispecies Biofilm Using Two Rotary Instrumentation Sequences

2020 ◽  
Vol 14 (01) ◽  
pp. 001-007
Author(s):  
Angelo Zavattini ◽  
Jonathan Cowie ◽  
Sadia Niazi ◽  
Massimo Giovarruscio ◽  
Salvatore Sauro ◽  
...  
Keyword(s):  
Statistical Analysis ◽  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Control Group ◽  
Multispecies Biofilm ◽  
Colony Forming Units ◽  
Mechanical Shaping ◽  
Different Levels

Abstract Objective The purpose of this research was to investigate the effect per se of two shaping and cleaning techniques on the reduction of an in vitro multispecies biofilm. Materials and Methods A total of 39 freshly extracted monoradicular teeth for periodontal reason were decoronated. Roots were sectioned longitudinally. After autoclaving, a specific stressed biofilm was grown on the root halves that were subsequently reassembled in a silicone index. Two treatments (n = 9 each)—RaCe (Schottlander; Letchworth Garden City, United Kingdom) and ProTaper Gold ( PTG; Dentsply Maillefer, Baillagues, Switzerland)—were tested; three noninstrumented samples served as a control group and three were rinsed with saline. Posttreatment samples were taken at three different levels of the root. Colony-forming units were counted after incubations. Additionally, three treatments (n = 5 each)—RaCe, PTG, and saline only—were evaluated under a confocal laser scanning microscope (CLSM). Statistical Analysis Statistical analysis was conducted using Tukey’s test and analysis of variance to evaluate the post-instrumentation bioburden. Results Both instrumentations were able to reduce the biofilm; however, differences were not present between them (p > 0.05). CLSM showed biofilm killing and disruption through mechanical shaping alone. Conclusions Intraradicular biofilm is reduced with mechanical shaping. There was no difference between RaCe and PTG systems in biofilm reduction despite differences in design, file sequence, and rotational speed.

scholarly journals Comparative Evaluation of Resin Dentin Interface using Universal and Total- Etch Adhesive Systems on Sound and Eroded Dentin: In Vitro Study

2021 ◽  
Author(s):  
Ghayathri Kanniappan ◽  
Padmini Hari ◽  
Ravikanth H. Jujare
Keyword(s):  
Laser Scanning ◽  
Artificial Saliva ◽  
Test Group ◽  
Confocal Laser ◽  
Control Group ◽  
Adhesive Systems ◽  
Hybrid Layer ◽  
Occlusal Surfaces ◽  
Self Etch

Abstract Objective The study aimed to compare the resin-dentin interface of sound and eroded dentin using universal and total-etch adhesive systems. Material and Methods Forty caries-free extracted human premolars were collected, and the occlusal surfaces were ground by using slow speed diamond disc with copious water supply until a flat superficial dentin was exposed. The test group underwent erosive cycle (n = 20), and another group (n = 20) was reserved for control group. Erosive protocol consisted of immersion in 1.23% citric acid for 1 minute every 12 hours and stored in artificial saliva. Both the control and eroded teeth were further subdivided (n = 10) for composite restoration by using either self-etch or total-etch systems. Then the tooth samples were sectioned longitudinally and observed under confocal laser scanning microscope at ×10 magnification to evaluate resin tag length and hybrid layer thickness. Statistical Analysis The data obtained were analyzed by using independent t-test. Results The highest mean value of the resin tag length and thickness of hybrid layer was observed with total-etch system in sound dentin group compared with other groups (p < 0.001). Conclusion The resin-dentin interface of sound dentin was found to be better than eroded dentin by using total-etch system. The resin-dentin interface of eroded dentin was superior to sound dentin by using self-etch adhesive system.

Vacuoles Induced in Mammalian Cells by Helicobacter Cytotoxin are Acidic Organelles

1990 ◽  
Vol 48 (3) ◽  
pp. 168-169
Author(s):  
M. H. Chestnut ◽  
C. E. Catrenich
Keyword(s):  
Acridine Orange ◽  
Mammalian Cells ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ulcer Disease ◽  
Gastroduodenal Disease ◽  
H Pylori

Helicobacter pylori is a non-invasive, Gram-negative spiral bacterium first identified in 1983, and subsequently implicated in the pathogenesis of gastroduodenal disease including gastritis and peptic ulcer disease. Cytotoxic activity, manifested by intracytoplasmic vacuolation of mammalian cells in vitro, was identified in 55% of H. pylori strains examined. The vacuoles increase in number and size during extended incubation, resulting in vacuolar and cellular degeneration after 24 h to 48 h. Vacuolation of gastric epithelial cells is also observed in vivo during infection by H. pylori. A high molecular weight, heat labile protein is believed to be responsible for vacuolation and to significantly contribute to the development of gastroduodenal disease in humans. The mechanism by which the cytotoxin exerts its effect is unknown, as is the intracellular origin of the vacuolar membrane and contents. Acridine orange is a membrane-permeant weak base that initially accumulates in low-pH compartments. We have used acridine orange accumulation in conjunction with confocal laser scanning microscopy of toxin-treated cells to begin probing the nature and origin of these vacuoles.

scholarly journals Effect of dissolution rate and subsequent ion release on cytocompatibility properties of borophosphate glasses

2019 ◽  
Vol 5 (1) ◽  
pp. 85-97
Author(s):  
Nusrat Sharmin ◽  
Mohammad S. Hasan ◽  
Md. Towhidul Islam ◽  
Chengheng Pang ◽  
Fu Gu ◽  
...  
Keyword(s):  
Dissolution Rate ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ion Release ◽  
Scanning Calorimetry ◽  
Cell Culture Study ◽  
Borophosphate Glasses ◽  
Mg63 Cells

AbstractPresent work explores the relationship between the composition, dissolution rate, ion release and cytocompatibility of a series of borophosphate glasses. While, the base glass was selected to be 40mol%P2O5-16mol%CaO-24mol%MgO-20mol%Na2O, three B2O3 modified glass compositions were formulated by replacing Na2O with 1, 5 and 10 mol% B2O3. Ion release study was conducted using inductively coupled plasma atomic emission spectroscopy (ICP-AES). The thermal scans of the glasses as determined by differential scanning calorimetry (DSC) revealed an increment in the thermal properties with increasing B2O3 content in the glasses. On the other hand, the dissolution rate of the glasses decreased with increasing B2O3 content. To identify the effect of boron ion release on the cytocompatibility properties of the glasses, MG63 cells were cultured on the surface of the glass discs. The in vitro cell culture study suggested that glasses with 5 mol% B2O3 (P40B5) showed better cell proliferation and metabolic activity as compares to the glasses with 10 mol% (P40B10) or with no B2O3 (P40B0). The confocal laser scanning microscopy (CLSM) images of live/dead stained MG63 cells attached to the surface of the glasses also revealed that the number of dead cells attached to P40B5 glasses were significantly lower than both P40B0 and P40B10 glasses.

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