scholarly journals The Amelogenin-Derived Peptide TVH-19 Promotes Dentinal Tubule Occlusion and Mineralization

Polymers ◽  
2021 ◽  
Vol 13 (15) ◽  
pp. 2473
Author(s):  
Xiu Peng ◽  
Sili Han ◽  
Kun Wang ◽  
Longjiang Ding ◽  
Zhenqi Liu ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Fluorescein Isothiocyanate ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Dentinal Tubule ◽  
X Ray ◽  
Electron Microscopy Analysis ◽  
Tubule Occlusion ◽  
Demineralized Dentin

In this study, the amelogenin-derived peptide, TVH-19, which has been confirmed to promote mineralization, was evaluated to derive its potential to induce dentinal tubule occlusion. The binding capability of fluorescein isothiocyanate (FITC)-labeled TVH-19 to the demineralized dentin surface was analyzed by confocal laser scanning microscopy (CLSM). Additionally, the sealing function of the peptide was studied through the remineralization of demineralized dentin in vitro. The adsorption results showed that TVH-19 could bind to the hydroxyapatite and demineralized dentin surfaces, especially to periodontal dentin. Scanning electron microscopy analysis further revealed that TVH-19 created mineral precipitates. The plugging rate in the TVH-19 group was higher than that in the PBS group. Moreover, energy-dispersive X-ray spectroscopy (EDX) results indicated that the calcium/phosphorus (Ca/P) ratio of the new minerals induced by TVH-19 was close to that of the hydroxyapatite. Attenuated total internal reflection-Fourier transform infrared (ATR-FTIR) spectrometry and X-ray diffraction (XRD) results indicated that the hydroxyapatite crystals formed via remineralization elongated the axial growth and closely resembled the natural dentin components. These findings indicate that TVH-19 can effectively promote dentin sealing by binding to the periodontal dentin, promoting mineral deposition, and reducing the space between the dentin tubules.

Internalized SP-A and lipid are differentially resecreted by type II pneumocytes

2000 ◽  
Vol 278 (3) ◽  
pp. L580-L590 ◽  
Author(s):  
Heide Wissel ◽  
Stefan Zastrow ◽  
Ekkehard Richter ◽  
Paul A. Stevens
Keyword(s):  
Laser Scanning ◽  
Protein A ◽  
Fluorescein Isothiocyanate ◽  
Surfactant Protein ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Type Ii Cells ◽  
Type Ii ◽  
Type Ii Pneumocytes

Biochemical and morphological assays were developed to study surfactant protein A (SP-A) and lipid resecretion kinetics by isolated type II cells in vitro. After a 10-min uptake period with SP-A (3 μg/106 cells) in combination with liposomes (60 μg/106 cells), the cells were allowed to resecrete. After 5 min of resecretion, only 21.7 ± 4.6% of the internalized SP-A remained intracellularly compared with 54 ± 2.9% of the lipids. Extracellular SP-A present during the resecretion period partially inhibited resecretion (SP-A, 36% at 5 min; lipid, ∼16% at 5 min). Lipid resecretion was also dependent on the SP-A concentration present during the uptake period. Although, as shown by confocal laser scanning microscopy, after a 10-min uptake period at 37°C, most of the fluorescein isothiocyanate-labeled SP-A and rhodamine-phosphatidylethanolamine-labeled lipids colocalized within the cells, after an additional 10 min of resecretion, both the strength of the fluorescence signals and the extent of colocalization had markedly decreased. These data indicate that internalized lipid and SP-A can be resecreted rapidly by type II cells, likely via different pathways.

scholarly journals Impact of Warm Vertical Compaction on the Sealing Ability of Calcium Silicate-Based Sealers: A Confocal Microscopic Evaluation

Materials ◽  
2021 ◽  
Vol 14 (2) ◽  
pp. 372
Author(s):  
Diana Eid ◽  
Etienne Medioni ◽  
Gustavo De-Deus ◽  
Issam Khalil ◽  
Alfred Naaman ◽  
...  
Keyword(s):  
Calcium Silicate ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Dentinal Tubule ◽  
Microscopic Evaluation ◽  
Significant Difference ◽  
Maximum Penetration ◽  
Warm Vertical Compaction

The aim of this in vitro study was to evaluate the dentinal tubule penetration of two calcium silicate-based sealers used in warm vertical compaction (WVC) obturation technique in comparison with the single cone (SC) technique by confocal laser scanning microscopy (CLSM). The null hypothesis was that both obturation techniques produced similar sealer penetration depths at 1 and 5 mm from the apex. Forty-four mandibular single-rooted premolars were randomly divided into four equally experimental groups (n = 10) and two control groups (n = 2) according to the type of sealer (Bio-C Angelus, Londrína, PR, Brazil or HiFlow Brasseler, Savannah, GA, USA) with either SC or WVC. The sealers were mixed with a fluorescent dye Rhodamine B (0.1%) to enable the assessment under the CLSM. All the specimens were sectioned horizontally at 1 and 5 mm from the apex. The maximum penetration depth was calculated using the ImageJ Software (ImageJ, NIH). Data were analyzed by Mann–Whitney U and Kruskal–Wallis tests (p < 0.05). A significant difference was shown between the four groups at 1 mm (p = 0.0116), whereas similar results were observed at 5 mm (p = 0.20). WVC allowed better diffusion for both sealers at 1 mm (p = 0.01) and 5 mm (p = 0.034). The maximum penetration of the Bio-C and HiFlow sealers was more important at 5 mm with the two obturation techniques. Within the limitations of this study, WVC enhanced the penetration of calcium silicate-based sealers into the dentinal tubules in comparison with the SC technique at both levels.

scholarly journals Laser Surface Modification of Powder Metallurgy-Processed Ti-Graphite Composite Which Can Enhance Cells’ Osteo-Differentiation

Materials ◽  
2021 ◽  
Vol 14 (20) ◽  
pp. 6067
Author(s):  
Peter Šugár ◽  
Barbora Ludrovcová ◽  
Marie Hubálek Kalbáčová ◽  
Jana Šugárová ◽  
Martin Sahul ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Distribution Curve ◽  
Surface Profile ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
In Vitro Tests ◽  
Nanosecond Laser ◽  
Graphite Composite ◽  
X Ray

The paper examines the surface functionalization of a new type of Ti-graphite composite, a dental biomaterial prepared by vacuum low-temperature extrusion of hydrogenated-dehydrogenated titanium powder mixed with graphite flakes. Two experimental surfaces were prepared by laser micromachining applying different levels of incident energy of the fiber nanosecond laser working at 1064 nm wavelength. The surface integrity of the machined surfaces was evaluated, including surface roughness parameters measurement by contact profilometry and confocal laser scanning microscopy. The chemical and phase composition were comprehensively evaluated by scanning electron microscopy, energy-dispersive X-ray spectroscopy and X-ray diffraction analyses. Finally, the in vitro tests using human mesenchymal stem cells were conducted to compare the influence of the laser processing parameters used on the cell’s cultivation and osteo-differentiation. The bioactivity results confirmed that the surface profile with positive kurtosis, platykurtic distribution curve and higher value of peaks spacing exhibited better bioactivity compared to the surface profile with negative kurtosis coefficient, leptokurtic distribution curve and lower peaks spacing.

scholarly journals Growth-Inhibitory Effect of Heparin on Babesia Parasites

2004 ◽  
Vol 48 (1) ◽  
pp. 236-241 ◽  
Author(s):  
Sabine Bork ◽  
Naoaki Yokoyama ◽  
Yuzuru Ikehara ◽  
Sanjay Kumar ◽  
Chihiro Sugimoto ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Fluorescein Isothiocyanate ◽  
Inhibitory Effect ◽  
Laser Scanning Microscopy ◽  
Babesia Bovis ◽  
Confocal Laser ◽  
Intracellular Parasites ◽  
Low Concentrations

ABSTRACT We examined the inhibitory effects of three heparins on the growth of Babesia parasites. The multiplication of Babesia bovis, B. bigemina, B. equi, and B. caballi in in vitro cultures and that of B. microti in vivo were significantly inhibited in the presence of heparins, as determined by light microscopy. Treatment with various concentrations of heparin showed complete clearance of the intracellular parasites. Interestingly, a higher percentage of abnormally multidividing B. bovis parasites was observed in the presence of low concentrations of heparin. Furthermore, fluorescein isothiocyanate-labeled heparin was preferably found on the surfaces of extracellular merozoites, as detected by confocal laser scanning microscopy. These findings indicate that the heparin covers the surfaces of babesial merozoites and inhibits their subsequent invasion of erythrocytes.

scholarly journals Physico-Chemical Properties and In Vitro Antifungal Evaluation of Samarium Doped Hydroxyapatite Coatings

Coatings ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 827 ◽  
Author(s):  
Steluta Carmen Ciobanu ◽  
Simona Liliana Iconaru ◽  
Daniela Predoi ◽  
Alina Mihaela Prodan ◽  
Mihai Valentin Predoi
Keyword(s):  
Photoelectron Spectroscopy ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Surface Composition ◽  
Sol Gel ◽  
Dip Coating ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
X Ray

Hydroxyapatite (HAp) and samarium doped hydroxyapatite, Ca10−xSmx(PO4)6(OH)2, xSm = 0.05, (5SmHAp), coatings were prepared by sol-gel process using the dip coating method. The stability of 5SmHAp suspension was evaluated by ultrasound measurements. Fourier transform infrared spectroscopy (FTIR) was used to examine the optical characteristics of HAp and 5SmHAp nanoparticles in suspension and coatings. The FTIR analysis revealed the presence of the functional groups specific to the structure of hydroxyapatite in the 5SmHAp suspensions and coatings. The morphology of 5SmHAp nanoparticles in suspension was evaluated by transmission electron microscopy (TEM). Moreover, scanning electron microscope (SEM) was used to evaluate the morphology of nanoparticle in suspension and the morphology of the surface on the coating. The SEM and TEM studies on 5SmHAp nanoparticles in suspension showed that our samples consist of nanometric particles with elongated morphology. The SEM micrographs of HAp and 5SmHAp coatings pointed out that the coatings are continuous and homogeneous. The surface morphology of the 5SmHAp coatings was also assessed by Atomic Force Microscopy (AFM) studies. The AFM results emphasized that the coatings presented the morphology of a uniformly deposited layer with no cracks and fissures. The crystal structure of 5SmHAp coating was characterized by X-ray diffraction (XRD). The surface composition of 5SmHAp coating was analyzed by X-ray photoelectron spectroscopy (XPS). The XRD and XPS analysis shown that the Sm3+ ions have been incorporated into the 5SmHAp synthesized material. The antifungal properties of the 5SmHAp suspensions and coatings were studied using Candida albicans ATCC 10231 (C. albicans) fungal strains. The quantitative results of the antifungal assay showed that colony forming unity development was inhibited from the early phase of adherence in the case of both suspensions and coatings. Furthermore, the adhesion, cell proliferation and biofilm formation of the C. albicans were also investigated by AFM, SEM and Confocal Laser Scanning Microscopy (CLSM) techniques. The results highlighted that the C. albicans adhesion and cell development was inhibited by the 5SmHAp coatings. Moreover, the data also revealed that the 5SmHAp coatings were effective in stopping the biofilm formation on their surface. The toxicity of the 5SmHap was also investigated in vitro using HeLa cell line.

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.

scholarly journals Cephalosporin nitric oxide-donor prodrug DEA-C3D disperses biofilms formed by clinical cystic fibrosis isolates of Pseudomonas aeruginosa

2019 ◽  
Vol 75 (1) ◽  
pp. 117-125 ◽  
Author(s):  
Odel Soren ◽  
Ardeshir Rineh ◽  
Diogo G Silva ◽  
Yuming Cai ◽  
Robert P Howlin ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Clinical Isolates ◽  
Nitric Oxide Donor ◽  
Confocal Laser ◽  
Broth Microdilution Method

Abstract Objectives The cephalosporin nitric oxide (NO)-donor prodrug DEA-C3D (‘DiEthylAmin-Cephalosporin-3′-Diazeniumdiolate’) has been shown to initiate the dispersal of biofilms formed by the Pseudomonas aeruginosa laboratory strain PAO1. In this study, we investigated whether DEA-C3D disperses biofilms formed by clinical cystic fibrosis (CF) isolates of P. aeruginosa and its effect in combination with two antipseudomonal antibiotics, tobramycin and colistin, in vitro. Methods β-Lactamase-triggered release of NO from DEA-C3D was confirmed using a gas-phase chemiluminescence detector. MICs for P. aeruginosa clinical isolates were determined using the broth microdilution method. A crystal violet staining technique and confocal laser scanning microscopy were used to evaluate the effects of DEA-C3D on P. aeruginosa biofilms alone and in combination with tobramycin and colistin. Results DEA-C3D was confirmed to selectively release NO in response to contact with bacterial β-lactamase. Despite lacking direct, cephalosporin/β-lactam-based antibacterial activity, DEA-C3D was able to disperse biofilms formed by three P. aeruginosa clinical isolates. Confocal microscopy revealed that DEA-C3D in combination with tobramycin produces similar reductions in biofilm to DEA-C3D alone, whereas the combination with colistin causes near complete eradication of P. aeruginosa biofilms in vitro. Conclusions DEA-C3D is effective in dispersing biofilms formed by multiple clinical isolates of P. aeruginosa and could hold promise as a new adjunctive therapy to patients with CF.

scholarly journals Use of electromagnetic stimulation on an Enterococcus faecalis biofilm on root canal treated teeth in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Beatriz H. D. Panariello ◽  
Justin K. Kindler ◽  
Kenneth J. Spolnik ◽  
Ygal Ehrlich ◽  
George J. Eckert ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Laser Scanning ◽  
Confocal Imaging ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Microscopy Imaging ◽  
Electromagnetic Stimulation ◽  
Root Canal Disinfection

AbstractRoot canal disinfection is of utmost importance in the success of the treatment, thus, a novel method for achieving root canal disinfection by electromagnetic waves, creating a synergistic reaction via electric and thermal energy, was created. To study electromagnetic stimulation (EMS) for the disinfection of root canal in vitro, single rooted teeth were instrumented with a 45.05 Wave One Gold reciprocating file. Specimens were sterilized and inoculated with Enterococcus faecalis ATCC 29,212, which grew for 15 days to form an established biofilm. Samples were treated with 6% sodium hypochlorite (NaOCl), 1.5% NaOCl 1.5% NaOCl with EMS, 0.9% saline with EMS or 0.9% saline. After treatments, the colony forming units (CFU) was determined. Data was analyzed by Wilcoxon Rank Sums Test (α = 0.05). One sample per group was scored and split for confocal laser scanning microscopy imaging. There was a significant effect with the use of NaOCl with or without EMS versus 0.9% saline with or without EMS (p = 0.012 and 0.003, respectively). CFUs were lower when using 0.9% saline with EMS versus 0.9% saline alone (p = 0.002). Confocal imaging confirmed CFU findings. EMS with saline has an antibiofilm effect against E. faecalis and can potentially be applied for endodontic disinfection.

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