scholarly journals Biological Response Induced in Primary Human Gingival Fibroblasts upon Exposure to Various Types of Injectable Astringent Retraction Agents

Materials ◽  
2021 ◽  
Vol 14 (8) ◽  
pp. 2081
Author(s):  
Danuta Nowakowska ◽  
Julita Kulbacka ◽  
Joanna Wezgowiec ◽  
Anna Szewczyk ◽  
Dagmara Baczynska ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Antioxidant Enzymes ◽  
Statistical Significance ◽  
Laser Scanning Microscopy ◽  
Human Gingival Fibroblasts ◽  
Aluminium Chloride ◽  
Confocal Laser ◽  
Rna Expression ◽  
Gingival Fibroblasts ◽  
Gingival Margin

Traditional chemo-mechanical retraction/displacement materials can impact the gingival margin tissues. This study was undertaken to analyze biological responses induced in human gingival fibroblasts (HGFs) upon application of injectable astringent-based agents used in the cordless retraction technique. HGFs were exposed to hemostatic agents (five gels, three pastes, and one foam) based on aluminium chloride, aluminium sulphate and ferric sulphate. Changes in cell viability and proliferation were evaluated using an MTT assay and a BrdU assay. The cytoskeleton structure organization (zyxin and F-actin) was visualized by confocal laser scanning microscopy. Oxidative stress was determined using the Griess Reagent System. The RNA expression levels of antioxidant enzymes were quantified by real-time RT-PCR. The statistical significance was evaluated using Student’s t-test and one-way ANOVA with post-hoc Tukey HSD test. The evaluated agents did not downregulate fibroblast viability or proliferation. No significant cytoskeleton reorganization was observed. Only one agent (Expasyl) induced oxidative stress, demonstrated by the increased level of nitrites. Incubation with the studied agents significantly increased the RNA expression of some antioxidant enzymes (SOD1, SOD3, GPX1). However, no significant influence on the expression of SOD2 and HMOX1 was detected. The injectable forms of chemical retraction agents revealed biocompatibility with HGFs, suggesting their potential clinical usefulness in gingival margin retraction.

Matrix expression and proliferation of primary gingival fibroblasts in a three-dimensional cell culture model

1999 ◽  
Vol 112 (17) ◽  
pp. 2823-2832 ◽  
Author(s):  
G. Hillmann ◽  
A. Gebert ◽  
W. Geurtsen
Keyword(s):  
Extracellular Matrix ◽  
Culture System ◽  
Cultured Cells ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Human Gingival Fibroblasts ◽  
Confocal Laser ◽  
Type I ◽  
Gingival Fibroblasts ◽  
Histological Techniques

The growth of cultured primary human gingival fibroblasts and the three-dimensional arrangement of the extracellular matrix in a polyester carrier system was investigated using various histological techniques. The results were compared with monolayer cultures. Collagen types I, III, V, and VI were investigated by conventional and fluorescence microscopy, scanning and transmission electron microscopy, and confocal laser scanning microscopy. Human gingival fibroblasts were obtained from tissue biopsies of five donors and were cultivated up to 5 weeks under three-dimensional culture conditions. The cells displayed an elongated, spindle-like or stellate morphology resembling the in vivo situation. Collagen type I revealed thick fiber bundles, and collagens type III and V were distributed as fine fibrils or small bundles throughout the culture system. Frequently, the fibers were oriented parallel to the long axis of the cells. Type VI collagen formed thin fibers and revealed a reticular pattern. In histological sections the cultured cells exhibited a morphology clearly different from that of cells cultured in monolayers. Their shape and spatial distribution resembled that of cells in tissue biopsies more closely. The culture system presented here promotes a dynamic model for performing studies for instance on the interactions of cultured cells with extracellular matrix molecules, on the pathogenesis of inflammatory processes or on the interactions with biomaterials, thus providing qualitative and quantitative information.

scholarly journals Potential role of nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase in apoptosis and oxidative stress

2001 ◽  
Vol 114 (9) ◽  
pp. 1643-1653 ◽  
Author(s):  
Z. Dastoor ◽  
J.L. Dreyer
Keyword(s):  
Oxidative Stress ◽  
Laser Scanning ◽  
Fluorescent Protein ◽  
Nuclear Translocation ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Apoptotic Cells ◽  
Transfected Cells ◽  
And Oxidative Stress

Recent studies indicating a role of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in apoptosis or oxidative stress has been reported. Using confocal laser-scanning microscopy, we have investigated the cellular distribution of GAPDH in central nervous system (CNS)-derived cells (neuroblastoma mNB41A3), in non-CNS derived cells (R6 fibroblast) and in an apoptosis-resistant Bcl2 overexpressing cell line (R6-Bcl2). Induction of apoptosis by staurosporine or MG132 and oxidative stress by H(2)O(2) or FeCN enhanced the nuclear translocation of endogenous GAPDH in all cell types, as detected by immunocytochemistry. In apoptotic cells, GAPDH expression is three times higher than in non-apoptotic cells. Consistent with a role for GAPDH in apoptosis, overexpression of a GAPDH-green fluorescent protein (GAPDH-GFP) hybrid increased nuclear import of GAPDH-GFP into transfected cells and the number of apoptotic cells, and made them more sensitive to agents that induce apoptosis. Bcl2 overexpression prevents nuclear translocation of GAPDH and apoptosis in untransfected cells, but not in transfected cells that overexpress GAPDH-GFP. Our observations indicate that nuclear translocation of GAPDH may play a role in apoptosis and oxidative stress, probably related to the activity of GAPDH as a DNA repair enzyme or as a nuclear carrier for pro-apoptotic molecules.

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

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
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.

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)

The Upregulation of Transglutaminase-2 by Cyclosporin A in Human Gingival Fibroblasts Is Augmented by Oxidative Stress

2013 ◽  
Vol 84 (10) ◽  
pp. 1469-1475 ◽  
Author(s):  
Shiuan-Shinn Lee ◽  
Chung-Hung Tsai ◽  
Yu-Hsiang Kuan ◽  
Fu-Mei Huang ◽  
Yu-Chao Chang
Keyword(s):  
Oxidative Stress ◽  
Cyclosporin A ◽  
Transglutaminase 2 ◽  
Human Gingival Fibroblasts ◽  
Gingival Fibroblasts

scholarly journals Cytotoxic and Genotoxic Effects of Composite Resins on Cultured Human Gingival Fibroblasts

Materials ◽  
2021 ◽  
Vol 14 (18) ◽  
pp. 5225
Author(s):  
Francesco De Angelis ◽  
Domitilla Mandatori ◽  
Valeria Schiavone ◽  
Francesco Paolo Melito ◽  
Silvia Valentinuzzi ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Metabolic Activity ◽  
Triethylene Glycol ◽  
Composite Resins ◽  
Dental Composite ◽  
Human Gingival Fibroblasts ◽  
Gingival Fibroblasts ◽  
Cell Metabolic Activity ◽  
Genotoxic Potential

The aim of the study was to evaluate the cytotoxic and genotoxic potential of five commercially available dental composite resins (CRs), investigating the effect of their quantifiable bisphenol-A-glycidyl-methacrylate (Bis-GMA) and/or triethylene glycol dimethacrylate (TEGDMA) release. Experiments were performed using the method of soaking extracts, which were derived from the immersion of the following CRs in the culture medium: Clearfil-Majesty-ES-2, GrandioSO, and Enamel-plus-HRi (Bis-GMA-based); Enamel-BioFunction and VenusDiamond (Bis-GMA-free). Human Gingival Fibroblasts (hGDFs) were employed as the cellular model to mimic in vitro the oral cavity milieu, where CRs simultaneously release various components. Cell metabolic activity, oxidative stress, and genotoxicity were used as cellular outcomes. Results showed that only VenusDiamond and Enamel-plus-HRi significantly affected the hGDF cell metabolic activity. In accordance with this, although no CR-derived extract induced a significantly detectable oxidative stress, only VenusDiamond and Enamel-plus-HRi induced significant genotoxicity. Our findings showed, for the CRs employed, a cytotoxic and genotoxic potential that did not seem to depend only on the actual Bis-GMA or TEGDMA content. Enamel-BioFunction appeared optimal in terms of cytotoxicity, and similar findings were observed for Clearfil-Majesty-ES-2 despite their different Bis-GMA/TEGDMA release patterns. This suggested that simply excluding one specific monomer from the CR formulation might not steadily turn out as a successful approach for improving their biocompatibility.

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