Antimicrobial Treatment of Dental Osseous Defects with Silver Doped Bioglass: Osteoblast Cell Response

2005 ◽  
Vol 284-286 ◽  
pp. 435-438 ◽  
Author(s):  
U. Lohbauer ◽  
G. Jell ◽  
Priya Saravanapavan ◽  
Julian R. Jones ◽  
Larry L. Hench
Keyword(s):  
Cell Viability ◽  
Culture Medium ◽  
Bactericidal Effect ◽  
Neutral Red ◽  
Antimicrobial Treatment ◽  
Mitochondrial Activity ◽  
Graft Material ◽  
Human Osteoblasts ◽  
Negative Effect ◽  
Silver Dissolution

In dentistry, chronic periodontitis often leads to bone resorption together with an increasing risk of bacteremia. Bioactive glass has found extensive application as dental graft material. A successful antimicrobial bactericidal effect has been shown from the introduction of Ag2O into the glass composition. In this study, the cytotoxicity of soluble silver, calcium and silica ions on primary human osteoblasts was investigated by measurements of mitochondrial activity and neutral red dye uptake. Silver concentrations of 4 - 6 ppm (1 mg/ml conc.) and 6 - 9 ppm (2 mg/ml conc.) have been measured in complete culture medium. It was found that the bioactive gel-glass extract with an initial concentration of 1 mg/ml (1mg glass per ml of culture medium) has no negative effect, whereas increased gel-glass concentration of 2 mg/ml seemed to have a toxic effect on the cell viability of human osteoblasts. It might be concluded that a reduction of the rate of silver dissolution from the bioactive gel-glass might preserve a maximum cell viability.

Indirect Cytotoxicity Evaluation of Silver Doped Bioglass Ag-S70C30 on Human Primary Keratinocytes

2005 ◽  
Vol 284-286 ◽  
pp. 431-434 ◽  
Author(s):  
U. Lohbauer ◽  
G. Jell ◽  
Priya Saravanapavan ◽  
Julian R. Jones ◽  
Larry L. Hench
Keyword(s):  
Cell Viability ◽  
Glass Powder ◽  
Culture Medium ◽  
Neutral Red ◽  
Epidermal Keratinocytes ◽  
Mitochondrial Activity ◽  
Human Epidermal Keratinocytes ◽  
Negative Effect ◽  
Healing Agent ◽  
Gel Glasses

Bioactive gel-glasses, such as the silver-doped Ag-S70C30 glass, can be used to modify the inflammatory response in a local body compartment such as in acne lesions and in nonhealing dermal wounds. In this study, the cytotoxicity of soluble silver, calcium and silica ions on human epidermal keratinocytes was investigated by measurements of mitochondrial activity (MTT assay) and neutral red dye uptake (NR assay). Ag-S70C30 extracts were prepared by soaking glass powder in complete culture medium at concentrations of 1 mg/ml and 2 mg/ml (mg of glass powder per ml of culture medium). Silver concentrations for both concentrations of approximately 1 ppm were detected by inductive coupled plasma analysis (ICP). No negative effect on the cell viability was measured for an initial gel-glass concentration of 1 mg/ml and for the two shortest extraction times at a concentration of 2 mg/ml. Based on the results from MTT/ NR assays, a pH rise of approximately one unit had no negative effect on the NHEK-A cell viability. This preliminary study on keratinocyte viability merits future investigations on silver bioglass as a novel antimicrobial wound healing agent.

scholarly journals Human Osteoblast Cell Behaviour on Titanium Discs Treated with Argon Plasma

Materials ◽  
2019 ◽  
Vol 12 (11) ◽  
pp. 1735 ◽  
Author(s):  
Carolina González-Blanco ◽  
María Rizo-Gorrita ◽  
Irene Luna-Oliva ◽  
María-Ángeles Serrera-Figallo ◽  
Daniel Torres-Lagares ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Cell Viability ◽  
Argon Plasma ◽  
Negative Control ◽  
Mitochondrial Activity ◽  
Cell Behaviour ◽  
Human Osteoblasts ◽  
Resistance To Stress ◽  
Viability Percentage ◽  
Cell Adhesion And Proliferation

(1) Background. Titanium is characterized by its biocompatibility and resistance to stress and fatigue. Treatment with argon plasma may favour growth of human osteoblasts with respect to cell adhesion and proliferation. The aim of this study was to analyse the behaviour of human osteoblasts (MG-63) on Grade IV and V titanium possessing a sand-blasted, acid-etched (SLA) surface. SLA is a widely used surface treatment to create micro- and macroretentions to enhance osteoconductive properties on the surface. (2) Methods. One group of each grade of titanium was decontaminated with argon plasma and compared. On each disc, 20 × 104 cells were cultivated for morphological analysis, study of cell viability (regarding a negative control [100% viability]) and mitochondrial energy balance. (3) Results. At 24 h titanium treated with SLA showed a higher percentage of cell viability (47.3 ± 8.1%) compared to titanium IV treated with argon plasma, which presented a percentage of 79.1 ± 1.1%. Grade V titanium treated with argon plasma presented a higher viability percentage 91.3 ± 3.0% whereas nontreated Grade V titanium presented 53.3 ± 4.0%. Cells cultivated on the surfaces with an argon-plasma treatment were enlarged in comparison to non-treated discs. The cells with smaller circularity with a greater spread and spindle shape were the ones cultivated on the Grade V titanium surface. Cells seeded on treated titanium IV and titanium V, treated or not, showed higher mitochondrial activity over nontreated titanium IV. (4) Conclusions. Cells cultivated on those Grade V titanium discs that were decontaminated with argon plasma presented higher levels of cell adhesion and proliferation, lower mitochondrial damage and a higher mean cell area compared to those not decontaminated with argon plasma.

scholarly journals Cytotoxicity assays as tools to assess water quality in the Sinos River basin

2015 ◽  
Vol 75 (2 suppl) ◽  
pp. 75-80 ◽  
Author(s):  
L Trintinaglia ◽  
E Bianchi ◽  
LB Silva ◽  
CA Nascimento ◽  
FR Spilki ◽  
...  
Keyword(s):  
Water Quality ◽  
River Basin ◽  
Water Samples ◽  
Culture Medium ◽  
Culture Media ◽  
Neutral Red ◽  
Mitochondrial Activity ◽  
Preparation Methods ◽  
Cytotoxicity Assays ◽  
Standard Culture

<p>Cytotoxicity assays using cell cultures may be an alternative to assess biological toxicity of surface waters and may help to improve the control of water quality. This study compared two methods to prepare culture media for the exposure of Hep-2 cells to water samples collected from the Rolante River, an important affluent of the Sinos River. The toxicity was evaluated using the MTT and neutral red assays. Two methods were used to prepare culture media. In method 1, the sample was diluted at 1:1, 1:10, 1:100, 1:1000, 1:10.000 (v/v, sample/medium) in a standard culture medium; in method 2, water samples were used as the solvent for the culture medium, which was prepared at concentrations of 100, 80, 60, 40 and 20%. Semi-confluent cultures were then exposed to the media test for 24 hours, and cytotoxicity was determined immediately using the MTT and NR assays. Mitochondrial activity (MTT) was significantly lower at all concentrations in both methods, except at 1:1000 in method 1. However, the lysosome viability (NR) results revealed cytotoxicity only in the 1:1 sample of method 1. Both culture preparation methods were efficient and sensitive to the MTT assay, but method 2 seemed to be more adequate for the NR assay. The Rolante River has cytotoxic contaminants to Hep-2 cells, which may be one of the explanations for the poor water quality of the Sinos River basin.</p>

Cytoprotective Effect of Tocotrienol-Rich Fraction and α-Tocopherol Vitamin E Isoforms Against Glutamate-Induced Cell Death in Neuronal Cells

2014 ◽  
Vol 84 (3-4) ◽  
pp. 0140-0151 ◽  
Author(s):  
Thilaga Rati Selvaraju ◽  
Huzwah Khaza’ai ◽  
Sharmili Vidyadaran ◽  
Mohd Sokhini Abd Mutalib ◽  
Vasudevan Ramachandran ◽  
...  
Keyword(s):  
Vitamin E ◽  
Cell Viability ◽  
Neuronal Cells ◽  
Positive Control ◽  
Post Treatment ◽  
Mitochondrial Activity ◽  
Before And After ◽  
Pre Treatment ◽  
Tocotrienol Rich Fraction ◽  
Major Mediator

Glutamate is the major mediator of excitatory signals in the mammalian central nervous system. Extreme amounts of glutamate in the extracellular spaces can lead to numerous neurodegenerative diseases. We aimed to clarify the potential of the following vitamin E isomers, tocotrienol-rich fraction (TRF) and α-tocopherol (α-TCP), as potent neuroprotective agents against glutamate-induced injury in neuronal SK-N-SH cells. Cells were treated before and after glutamate injury (pre- and post-treatment, respectively) with 100 - 300 ng/ml TRF/α-TCP. Exposure to 120 mM glutamate significantly reduced cell viability to 76 % and 79 % in the pre- and post-treatment studies, respectively; however, pre- and post-treatment with TRF/α-TCP attenuated the cytotoxic effect of glutamate. Compared to the positive control (glutamate-injured cells not treated with TRF/α-TCP), pre-treatment with 100, 200, and 300 ng/ml TRF significantly improved cell viability following glutamate injury to 95.2 %, 95.0 %, and 95.6 %, respectively (p < 0.05).The isomers not only conferred neuroprotection by enhancing mitochondrial activity and depleting free radical production, but also increased cell viability and recovery upon glutamate insult. Our results suggest that vitamin E has potent antioxidant potential for protecting against glutamate injury and recovering glutamate-injured neuronal cells. Our findings also indicate that both TRF and α-TCP could play key roles as anti-apoptotic agents with neuroprotective properties.

In vitro Growth Pattern of Primary Human Osteoblasts on Calcium Phosphate- and Polymethylmethacrylate-Based Bone Cement

2017 ◽  
Vol 58 (5-6) ◽  
pp. 216-226
Author(s):  
Johannes Schauwecker ◽  
Mark Bock ◽  
Florian Pohlig ◽  
Heinz Mühlhofer ◽  
Jutta Tübel ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Calcium Phosphate ◽  
Cell Viability ◽  
Proliferation Rate ◽  
Control Analysis ◽  
Implant Loosening ◽  
Human Osteoblasts ◽  
Apoptosis Rate ◽  
Primary Human Osteoblasts

Background/Purpose: Polymethylmethacrylate (PMMA) and calcium phosphate (Ca-P) cements are widely used for arthroplasty surgery and augmentation of bone defects. However, aseptic implant loosening in absence of wear-induced osteolysis indicates an unfavourable interaction between the cement surface and human osteoblasts. Our underlying hypothesis is that cement surfaces directly modify cell viability, proliferation rate, and cell differentiation. Methods: To test this hypothesis, we examined primary human osteoblasts harvested from six individuals. These cells were pooled and subsequently seeded directly on cement pellets prepared from Palacos® R, Palacos® R+G, and Norian® Drillable cements. After incubation for 24 and 72 h, cell viability, proliferation rate, apoptosis rate, and cell differentiation were analysed. Results: Upon cultivation of human osteoblasts on cement surfaces, we observed a significantly reduced cell viability and DNA content compared to the control. Analysis of the apoptosis rate revealed an increase for cells on Palacos R and Norian Drillable, but a significant decrease on Palacos R+G compared to the control. Regarding osteogenic differentiation, significantly lower values of alkaline phosphatase enzyme activity were identified for all cement surfaces after 24 and 72 h compared to cultivation on tissue culture plastic, serving as control. Conclusions: In summary, these data suggest a limited biocompatibility of both PMMA and Ca-P cements, necessitating further research to reduce unfavourable cell-cement interactions and consequently extend implant survival.

scholarly journals Biological characterization of implant surfaces - in vitro study

2015 ◽  
Vol 44 (4) ◽  
pp. 195-199 ◽  
Author(s):  
Priscilla Barbosa Ferreira Soares ◽  
Camilla Christian Gomes Moura ◽  
Huberth Alexandre da Rocha Júnior ◽  
Paula Dechichi ◽  
Darceny Zanetta-Barbosa
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Viability ◽  
Total Protein ◽  
Cell Attachment ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Serum Total Protein ◽  
Trypan Blue Exclusion ◽  
Experimental Surface

<title>Abstract</title><sec><title>Objective</title><p>Evaluate the biological performance of titanium alloys grade IV under different surface treatments: sandblasting and double etching (Experimental surface 1; Exp1, NEODENT); surface with wettability increase (Experimental surface 2; Exp2, NEODENT) on response of preliminary differentiation and cell maturation.</p></sec><sec><title>Material and method</title><p>Immortalized osteoblast cells were plated on Exp1 and Exp2 titanium discs. The polystyrene plate surface without disc was used as control group (C). Cell viability was assessed by measuring mitochondrial activity (MTT) at 4 and 24 h (n = 5), cell attachment was performed using trypan blue exclusion within 4 hours (n = 5), serum total protein and alkaline phosphatase normalization was performed at 4, 7 and 14 days (n = 5). Data were analyzed using one-way ANOVA and Tukey test.</p></sec><sec><title>Result</title><p>The values of cell viability were: 4h: C– 0.32±0.01<sup>A</sup>; Exp1– 0.34±0.08<sup>A</sup>; Exp2– 0.29±0.03<sup>A</sup>. 24h: C– 0.43±0.02<sup>A</sup>; Exp1– 0.39±0.01<sup>A</sup>; Exp2– 0.37±0.03<sup>A</sup>. The cell adhesion counting was: C– 85±10<sup>A</sup>; Exp1- 35±5<sup>B</sup>; Exp2– 20±2<sup>B</sup>. The amounts of serum total protein were 4d: C– 40±2<sup>B</sup>; Exp1– 120±10<sup>A</sup>; Exp2– 130±20<sup>A</sup>. 7d: C– 38±2<sup>B</sup>; Exp1– 75±4<sup>A</sup>; Exp2– 70±6<sup>A</sup>. 14 d: C– 100±3<sup>A</sup>; Exp1– 130±5<sup>A</sup>; Exp2– 137±9<sup>A</sup>. The values of alkaline phosphatase normalization were: 4d: C– 2.0±0.1<sup>C</sup>; Exp1– 5.1±0.8<sup>B</sup>; Exp2– 9.8±2.0<sup>A</sup>. 7d: C– 1.0±0.01<sup>C</sup>; Exp1– 5.3±0.5<sup>A</sup>; Exp2– 3.0±0.3<sup>B</sup>. 14 d: C– 4.1±0.3<sup>A</sup>; Exp1– 4.4±0.8<sup>A</sup>; Exp2– 2.2±0.2<sup>B</sup>. Different letters related to statistical differences.</p></sec><sec><title>Conclusion</title><p>The surfaces tested exhibit different behavior at dosage of alkaline phosphatase normalization showing that the Exp2 is more associated with induction of cell differentiation process and that Exp1 is more related to the mineralization process.</p></sec>

Monoisopropylglutathione ester protects A549 cells from the cytotoxic effects of sulphur mustard

1997 ◽  
Vol 16 (11) ◽  
pp. 636-644 ◽  
Author(s):  
Christopher D Lindsay ◽  
Joy L Hambrook ◽  
Alison F Lailey
Keyword(s):  
Cell Viability ◽  
Cell Cultures ◽  
A549 Cell ◽  
High Performance ◽  
Gentian Violet ◽  
A549 Cells ◽  
Rapid Onset ◽  
Neutral Red ◽  
Biochemical Basis ◽  
Sulphur Mustard

1 The A549 cell line was used to assess the toxicity of sulphur mustard (HD), using gentian violet (GV) and neutral red (NR) dyes as indicators of cell viability. It was found that exposure to concentrations in excess of 40 ?M HD resulted in a rapid onset of toxicity. 2 The ability of monoisopropylglutathione ester (MIPE) to protect A549 cells against the effects of a 100 ?M challenge dose ofHD was determined using the NR and GV assays. It was found that MIPE (8 mM) could protect cells against the effects ofHD though MIPE had to be present at the time of HD challenge. Cultures protected with MIPE were two times more viable than HD exposed cells 48 h after HD challenge when using the GV and NR assays to assess viability. Observations by phase contrast microscopy of NR and GV stained cultures confirmed these findings. Addition of MIPE after previously exposing the A549 cultures to HD (for up to 5 min) maintained cell viability at 72% compared to 37% for unprotected cultures, after which time viability fell significantly so that at 10 min there was no difference in viability between the MIPE treated and untreated cultures. 3 Pretreating A549 cultures with MIPE for 1 h followed by its removal prior to HD challenge did not maintain cell viability. Treatment of cultures with HD for 1 h followed by addition of MIPE did not maintain the viability of the cultures, thus the window within which it was possible for MIPE to rescue cell cultures from the effects of HD was of short duration. 4 High performance liquid chromatography was used to determine the biochemical basis of the actions of MIPE. It was found that whilst intracellular levels of cysteine were increased up to 40-fold following treatment of A549 cell cultures with MIPE, levels of reduced glutathione did not rise. The lack of protection seen in cultures pretreated with MIPE for 1 h prior to HD exposure suggests that raising intracellular cysteine levels was not an effective strategy for protecting cells from the effects of HD. The protection observed is probably due to extra cellular inactivation of HD by MIPE.

scholarly journals Selection of Trichoderma spp. strains for the control of Sclerotinia sclerotiorum in soybean

2017 ◽  
Vol 52 (12) ◽  
pp. 1140-1148 ◽  
Author(s):  
Patrícia Elias Haddad ◽  
Luis Garrigós Leite ◽  
Cleusa Maria Mantovanello Lucon ◽  
Ricardo Harakava
Keyword(s):  
Sclerotinia Sclerotiorum ◽  
Culture Medium ◽  
Harmful Effect ◽  
Soybean Seeds ◽  
Trichoderma Spp ◽  
Negative Effect ◽  
Soybean Plants ◽  
Selection Of

Abstract: The objective of this work was to evaluate, in vitro and in vivo, the potential of Trichoderma spp. strains to control Sclerotinia sclerotiorum in soybeans (Glycine max) and to perform the molecular identification of the best perfoming strains. The effect of 120 strains of Trichoderma spp. on the viability of S. sclerotiorum sclerotia was evaluated in vitro through immersion in suspension of conidia from the antagonists and plating in culture medium. The best performing strains were evaluated in vivo, in a greenhouse, for control of the pathogen inoculated on 'Pintado' soybean seeds and plants. Of the 120 strains tested in vitro, 22 strains of Trichoderma spp. caused 100% inhibition of sclerotia germination. In the greenhouse, five strains inhibited the negative effect of the pathogen on seed germination and two strains increased in up to 67% plant dry matter. The best performing strains were identified as T. koningiopsis (3 strains), T. asperelloides (3), T. atroviride (2), and T. virens (1). Trichoderma strains are able to protect soybean plants from the harmful effect of S. sclerotiorum and, at the same time, they can promote the growth of the aerial part in greenhouse conditions.

Sign in / Sign up

Export Citation Format

Share Document