Candida albicans Increases Dentine Demineralization Provoked by Streptococcus mutans Biofilm

2018 ◽  
Vol 53 (3) ◽  
pp. 322-331 ◽  
Author(s):  
Aline A. Sampaio ◽  
Samilly E. Souza ◽  
Antônio P. Ricomini-Filho ◽  
Altair A. Del Bel Cury ◽  
Yuri W. Cavalcanti ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Streptococcus Mutans ◽  
Laser Scanning ◽  
Surface Hardness ◽  
Laser Scanning Microscopy ◽  
Broth Culture ◽  
Extracellular Polysaccharides ◽  
Confocal Laser ◽  
Ph Indicator ◽  
Transmission Electron

Streptococcus mutans are considered the most cariogenic bacteria, but it has been suggested that Candida albicans could increase their cariogenicity. However, the effect of this dual-species microorganisms’ combination on dentine caries has not been experimentally evaluated. Biofilms of C. albicans, S. mutans and C. albicans + S. mutans (n = 12/biofilm) were grown in ultra-filtered tryptone yeast extract broth culture medium for 96 h on root dentine slabs of known surface hardness and exposed 8 times per day for 3 min to 10% sucrose. The medium was changed 2 times per day (after the 8 cariogenic challenges and after the overnight period of famine), and aliquots were analyzed to determinate the pH (indicator of biofilm acidogenicity). After 96 h, the biofilms were collected to determine the wet weight, colony-forming units, and the amounts of extracellular polysaccharides (soluble and insoluble). Dentine demineralization was assessed by surface hardness loss (% SHL). The architecture of the biofilms was analyzed by confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). Data were analyzed by ANOVA followed by Tukey’s test (α = 0.05). The dual-species C. albicans + S. mutans biofilm provoked higher % SHL on dentine (p < 0.05) than the S. mutans and C. albicans biofilm. This was supported by the results of biofilm acidogenicity and the amounts of soluble (6.4 ± 2.14 vs. 4.0 ± 0.94 and 1.9 ± 0.97, respectively) and insoluble extracellular polysaccharides (24.9 ± 9.22 vs. 18.9 ± 5.92 and 0.7 ± 0.48, respectively) (p < 0.05). The C. albicans biofilm alone presented low cariogenicity. The images by CLSM and TEM, respectively, suggest that the C. albicans + S. mutans biofilm is more voluminous than the S. mutans biofilm, and S. mutans cells interact with C. albicans throughout polysaccharides from the biofilm matrix. These findings show that C. albicans enhances the cariogenic potential of the S. mutans biofilm, increasing dentine demineralization.

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 The vicK gene of Streptococcus mutans mediates its cariogenicity via exopolysaccharides metabolism

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Yalan Deng ◽  
Yingming Yang ◽  
Bin Zhang ◽  
Hong Chen ◽  
Yangyu Lu ◽  
...  
Keyword(s):  
Dental Caries ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Single Species ◽  
Laser Scanning Microscopy ◽  
Gas Chromatography Mass Spectrometry ◽  
Extracellular Polysaccharides ◽  
Confocal Laser

AbstractStreptococcus mutans (S. mutans) is generally regarded as a major contributor to dental caries because of its ability to synthesize extracellular polysaccharides (EPS) that aid in the formation of plaque biofilm. The VicRKX system of S. mutans plays an important role in biofilm formation. The aim of this study was to investigate the effects of vicK gene on specific characteristics of EPS in S. mutans biofilm. We constructed single-species biofilms formed by different mutants of vicK gene. Production and distribution of EPS were detected through atomic force microscopy, scanning electron microscopy and confocal laser scanning microscopy. Microcosmic structures of EPS were analyzed by gel permeation chromatography and gas chromatography-mass spectrometry. Cariogenicity of the vicK mutant was assessed in a specific pathogen-free rat model. Transcriptional levels of cariogenicity-associated genes were confirmed by quantitative real-time polymerase chain reaction. The results showed that deletion of vicK gene suppressed biofilm formation as well as EPS production, and EPS were synthesized mostly around the cells. Molecular weight and monosaccharide components underwent evident alterations. Biofilms formed in vivo were sparse and contributed a decreased degree of caries. Moreover, expressional levels of genes related to EPS synthesis were down-regulated, except for gtfB. Our report demonstrates that vicK gene enhances biofilm formation and subsequent caries development. And this may due to its regulations on EPS metabolism, like synthesis or microcosmic features of EPS. This study suggests that vicK gene and EPS can be considered as promising targets to modulate dental caries.

scholarly journals Effect of LongZhang Gargle on Dual-Species Biofilm of Candida albicans and Streptococcus mutans

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Jinglei Gong ◽  
Die Hu ◽  
Jinzhi He ◽  
Ling Zou ◽  
Zhu Chen ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Streptococcus Mutans ◽  
Hot Spot ◽  
Fungal Spores ◽  
Cell Activity ◽  
Single Species ◽  
Laser Scanning Microscopy ◽  
Oral Disease ◽  
Extracellular Polysaccharides ◽  
Confocal Laser

Bioactive natural products have become a hot spot for oral disease treatments. At the present study, LongZhang Gargle was investigated for its effects on single-species biofilms of Candida albicans and dual-species biofilms of Candida albicans and Streptococcus mutans. Two different models of single and dual-species biofilms were grown in YNBB medium under appropriate conditions. Biofilm biomass, biofilm architecture, and cell activity in biofilms were assessed using Crystal Violet Staining, MTT, scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). Significant reductions of biofilm biomass and fungus activity were obtained when treated with LongZhang Gargle at 2% ( P < 0.05 ), 4% ( P < 0.05 ), and 8% ( P < 0.05 ) in single-species biofilms of C. albicans, and at 4% ( P < 0.05 ) and 8% ( P < 0.05 ) in double-species biofilms. Suppression of density, thickness, and the proportion of hyphae and fungal spores were obtained under SEM and CLSM. In conclusion, LongZhang Gargle affects single and dual-species biofilms by inhibiting biofilm biomass, cell activity, and formation of hyphae, but it does not affect the production of Extracellular polysaccharides (EPS). We speculate that LongZhang Gargle would be a promising natural drug, which can be used in treatment against C. albicans and S. mutans in oral diseases.

Contribution of Microscopy to a Better Knowledge of the Biology ofGiardia lamblia

2004 ◽  
Vol 10 (5) ◽  
pp. 513-527 ◽  
Author(s):  
Wanderley de Souza ◽  
Adriana Lanfredi-Rangel ◽  
Loraine Campanati
Keyword(s):  
Golgi Complex ◽  
Laser Scanning ◽  
Diarrheal Disease ◽  
Freeze Fracture ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Enzyme Cytochemistry ◽  
Morphological Studies ◽  
Transmission Electron ◽  
Microscopic Techniques

Giardia lambliais a flagellated protozoan of great medical and biological importance. It is the causative agent of giardiasis, one of the most prevalent diarrheal disease both in developed and third-world countries. Morphological studies have shown thatG. lambliadoes not present structures such as peroxisomes, mitochondria, and a well-elaborated Golgi complex. In this review, special emphasis is given to the contribution made by various microscopic techniques to a better knowledge of the biology of the protozoan. The application of video microscopy, immunofluorescence confocal laser scanning microscopy, and several techniques associated with transmission electron microscopy (thin section, enzyme cytochemistry, freeze-fracture, deep-etching, fracture-flip) to the study of the cell surface, peripheral vesicles, endoplasmic reticulum–Golgi complex system, and of the encystation vesicles found in trophozoites and during the process of trophozoite-cyst transformation are discussed.

Microtubule targeted therapeutics loaded polymeric assembled nanospheres for potentiation of antineoplastic activity

2016 ◽  
Vol 186 ◽  
pp. 45-59 ◽  
Author(s):  
Radhika Poojari ◽  
Rohit Srivastava ◽  
Dulal Panda
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Laser Scanning ◽  
Present Approach ◽  
Laser Scanning Microscopy ◽  
Microtubule Assembly ◽  
Confocal Laser ◽  
Cancer Cell Proliferation ◽  
Transmission Electron ◽  
Hepatocarcinoma Cells

Polymeric nanoassemblies represent an attractive strategy for efficient cellular internalization of microtubule targeted anticancer drugs. Using dynamic light scattering, zeta potential, transmission electron microscopy and scanning electron microscopy, the physical properties and surface morphology of microtubule-binding PEGylated PLGA assembled nanospheres (100–200 nm) were analyzed. The present approach leads to strong internalization as observed by confocal laser scanning microscopy and transmission electron microscopy in hepatocarcinoma cells. The effect of these nanoassemblies on microtubules and mitosis were explored using immunofluorescence microscopy. The effects of these nanoassemblies on cancer cell proliferation and cell death revealed their antitumor enhancing effects. Perturbation of the microtubule assembly, mitosis and nuclear modulations potentiated the antineoplastic effects delivered via nanospheres in hepatocarcinoma cells. The extensive biomolecular and physical characterizations of the synthesized nanoassemblies will help to design potent therapeutic materials and the present approach can be applied to deliver microtubule-targeted drugs for liver cancer therapy.

scholarly journals Macropinocytosis as a Mechanism of Entry into Primary Human Urethral Epithelial Cells by Neisseria gonorrhoeae

2000 ◽  
Vol 68 (3) ◽  
pp. 1696-1699 ◽  
Author(s):  
Michael K. Zenni ◽  
Peter C. Giardina ◽  
Hillery A. Harvey ◽  
Jianqiang Shao ◽  
Margaret R. Ketterer ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Laser Scanning ◽  
Actin Polymerization ◽  
Kinase Inhibitors ◽  
Fluorescein Isothiocyanate ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Transmission Electron ◽  
Microscopy Analysis ◽  
Phosphoinositide 3 Kinase

ABSTRACT Gonococcal entry into primary human urethral epithelial cells (HUEC) can occur by macropinocytosis. Scanning and transmission electron microscopy revealed lamellipodia surrounding gonococci, and confocal laser scanning microscopy analysis showed organisms colocalized with M r 70,000 fluorescein isothiocyanate-labeled dextran within the cells. Phosphoinositide 3-kinase inhibitors and an actin polymerization inhibitor prevented macropinocytic entry of gonococci into HUEC.

Soft Tissue and Epithelial Models

1999 ◽  
Vol 13 (1) ◽  
pp. 57-66 ◽  
Author(s):  
J.A. Jansen ◽  
E.T. Den Braber ◽  
X.F. Walboomers ◽  
J.E. De Ruijter
Keyword(s):  
Laser Scanning ◽  
Analytical Techniques ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Good Tool ◽  
Oral Implants ◽  
Body Tissues ◽  
Transmission Electron ◽  
Gingival Cells

The applicability of a biomaterial for the manufacturing of oral implants is determined by its physicochemical and geometric surface properties. Research, therefore, is concerned with the cellular reactions that occur when an implant material comes into contact with body tissues. For permucosal oral implants, this involves both the reaction of bone and gingival cells. In vitro cell culturing-including the use of various analytical techniques like light microscopy, scanning and transmission electron microscopy, confocal laser scanning microscopy, and digital image analysis-is a good tool whereby investigators can obtain more insight into the relevant components of implant-tissue adhesion. In the current overview, the role of cell models in oral implant research is discussed, specifically with reference to responses of epithelial cells and fibroblasts.

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