scholarly journals Experimental in vitro Comparative Study of Chromovitectomy Agents Cyto- and Phototoxicity

2020 ◽  
Vol 17 (3) ◽  
pp. 473-480
Author(s):  
N. M. Kislitsyna ◽  
S. V. Novikov ◽  
N. V. Perova ◽  
S. V. Kolesnik ◽  
A. I. Kolesnik ◽  
...  
Keyword(s):  
Safety Profile ◽  
Cytotoxic Effect ◽  
Vitreous Body ◽  
Fibroblast Cell ◽  
Negative Control ◽  
Mouse Fibroblast ◽  
Test Plate ◽  
Mouse Fibroblasts ◽  
Nih 3T3

Intravitreal use of vital dyes in combination with the action of endoillumination can induce a cyto- and phototoxic effect on posterior eye segment structures. The search for a staining agent with a maximum safety profile to retinal structures, intensively and selectively coloring vitreous body and vitreoretinal interface structure, remains relevant.Objective: to determine comparative viability of NIH / 3T3 mouse fibroblast cell culture with traditional agents for chromovitrectomy and “Vitreocontrast” suspension with and without endovitreal illumination.Materials and methods. NIH / 3T3 mouse fibroblast cultures contacted with agents for chromovitrectomy (MembraneBlue® Dual, Triamcinolone acetonide, “Vitreocontrast” suspension) and the corresponding controls in a volume of 50 μl / well. The test plate was irradiated with a Photon II illuminator (Synergetics, USA), working distance of 5 mm. The control tablet with the introduced preparations was not exposed to light. Next, the cells were washed and incubated, after which the morphology and lysis of the cells, as well as the number of proliferating relatively negative control of fibroblasts, were evaluated using the vital dye PrestoBlue Cell Viability Reagent. Negative control was the complete growth medium for the cultivation of mouse fibroblasts of the NIH / 3T3 line. The results of the cytotoxic reaction of a culture of mouse fibroblasts of the NIH / 3T3 line were interpreted using the table “The degree of cell response”.Results. Studies have shown that exposure to a source of endovitual illumination does not affect the cytotoxic effect of TA suspension and MembraneBlue® Dual dye. The TA suspension, both after light source and without it, has a moderate cytotoxic effect, and MembraneBlue® Dual has no cytotoxic effect on the culture of mouse fibroblasts of the NIH / 3T3 strain. Without light, “Vitreocontrast” suspension does not have cytotoxic effect on mouse fibroblasts culture NIH / 3T3 line. Light irradiation for 1 h increases the cytotoxicity of “Vitreocontrast” suspension to the level of unsharp cytotoxicity allowed by ISO Standard 10993-5-2011.Conclusion. The safety profile of MembraneBlue® Dual and “Vitreocontrast” suspension allows them to be recommended for use in endovitreal surgery. The cyto- and phototoxicity demonstrated in the experiment with TA suspension can reduce the functional outcomes of retinal surgery. 

Survival and Function of Fibroblasts Stored as Stock Cultures at a Non-freezing Temperature

1993 ◽  
Vol 21 (2) ◽  
pp. 206-209
Author(s):  
Anders H. G. Andrén ◽  
Anders P. Wieslander
Keyword(s):  
Cell Growth ◽  
Cell Line ◽  
Cell Lines ◽  
Fibroblast Cell ◽  
Freezing Temperature ◽  
Mouse Fibroblast ◽  
Toxic Response ◽  
Normal Manner ◽  
And Function

Cytotoxicity, measured as inhibition of cell growth of cultured cell lines, is a widely used method for testing the safety of biomaterials and chemicals. One major technical disadvantage with this method is the continuous routine maintenance of the cell lines. We decided to investigate the possibility of storing stock cultures of fibroblasts (L-929) in an ordinary refrigerator as a means of reducing the routine workload. Stock cultures of the mouse fibroblast cell line L-929 were prepared in plastic vials with Eagle's minimum essential medium. The vials were stored in a refrigerator at 4–10°C for periods of 7–31 days. The condition of the cells after storage was determined as cell viability, cell growth and the toxic response to acrylamide, measured as cell growth inhibition. We found that the L-929 cell line can be stored for 2–3, weeks with a viabilty > 90% and a cell growth of about 95%, compared to L-929 cells grown and subcultured in the normal manner. The results also show that the toxic response to acrylamide, using refrigerator stored L-929 cells, corresponds to that of control L-929 cells. We concluded that it is possible to store L-929 cells in a refrigerator for periods of up to 3 weeks and still use the cells for in vitro cytotoxic assays.

scholarly journals A Proposed Fabrication Method of Novel PCL-GEL-HAp Nanocomposite Scaffolds for Bone Tissue Engineering Applications

2010 ◽  
Vol 19 (4) ◽  
pp. 096369351001900 ◽  
Author(s):  
A. Hamlekhan ◽  
M. Mozafari ◽  
N. Nezafati ◽  
M. Azami ◽  
H. Hadipour
Keyword(s):  
Tissue Engineering ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Mechanical Test ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
Fabrication Method ◽  
Engineering Applications ◽  
Poly Caprolactone

In this study, poly(∊-caprolactone) (PCL), gelatin (GEL) and nanocrystalline hydroxyapatite (HAp) was applied to fabricate novel PCL-GEL-HAp nanaocomposite scaffolds through a new fabrication method. With the aim of finding the best fabrication method, after testing different methods and solvents, the best method and solvents were found, and the nanocomposites were prepared through layer solvent casting combined with freeze-drying. Acetone and distillated water were used as the PCL and GEL solvents, respectively. The mechanical test showed that the increasing of the PCL weight through the scaffolds caused the improvement of the final nanocomposite mechanical behavior due to the increasing of the ultimate stress, stiffness and elastic modulus (8 MPa for 0% wt PCL to 23.5 MPa for 50% wt PCL). The biomineralization investigation of the scaffolds revealed the formation of bone-like apatite layers after immersion in simulated body fluid (SBF). In addition, the in vitro cytotoxity of the scaffolds using L929 mouse fibroblast cell line (ATCC) indicated no sign of toxicity. These results indicated that the fabricated scaffold possesses the prerequisites for bone tissue engineering applications.

Sodium affinity of brain Na(+)-K(+)-ATPase is dependent on isozyme and environment of the pump

1990 ◽  
Vol 258 (5) ◽  
pp. C803-C811 ◽  
Author(s):  
J. L. Brodsky ◽  
G. Guidotti
Keyword(s):  
Plasma Membranes ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
Synaptic Plasma Membranes ◽  
Apparent Dissociation Constant ◽  
Low Sodium ◽  
Physiological Relevance ◽  
Mouse Fibroblast Cell Line

The sodium affinities for the two forms of the Na(+)-K(+)-ATPase in brain were characterized. To mimic physiological conditions, synaptosomes, which are pinched off presynaptic nerve termini, were used. Examination of the pump in vitro was performed by preparing synaptic plasma membranes (SPMs). It was first shown that synaptosomes contain the two forms of the Na(+)-K(+)-ATPase, alpha 1 and alpha 2, and that these forms have markedly different affinities for the inhibitory cardiac glycoside ouabain. The apparent dissociation constant (K0.5) of alpha 1 for sodium changed from 12 to 9 mM when going from synaptosomes to membranes. For alpha 2, however, a shift from 36 to 12.5 mM was evident. The conclusion is that in vivo alpha 2 exists as a low sodium affinity species but can be altered to a high-affinity form simply by vesicle disruption. By comparison, the Na(+)-K(+)-ATPase from the mouse fibroblast cell line, 3T3-F442A cells, expressed only the alpha 1-isozyme, as shown by immunoblotting and by measurement of its ouabain and sodium affinities. The physiological relevance of these observations is also presented.

scholarly journals The Use of Xanthan Gum as Vaccine Adjuvant: An Evaluation of Immunostimulatory Potential in BALB/c Mice and Cytotoxicity In Vitro

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Rodrigo Andrade Schuch ◽  
Thaís Larré Oliveira ◽  
Thaís Farias Collares ◽  
Leonardo Garcia Monte ◽  
Guilherme Roig Inda ◽  
...  
Keyword(s):  
Immune Response ◽  
Xanthan Gum ◽  
In Vitro Cytotoxicity ◽  
Mouse Fibroblast ◽  
Vaccine Adjuvant ◽  
Iodide Uptake ◽  
Adjuvant Activity ◽  
Cytotoxicity Effects ◽  
Nih 3T3

The successful production of new, safe, and effective vaccines that generate immunological memory is directly related to adjuvant feature, which is responsible for increasing and/or modulating the immune response. Several compounds display adjuvant activity, including carbohydrates. These compounds play important roles in the immune response, as well as having biocompatible properties in vaccine formulations. One such carbohydrate is xanthan gum, a polysaccharide that is produced by the plant-pathogenic bacterium Xanthomonas spp., which has adjuvant attributes. This study evaluated the immune response induced by xanthan gum associated with ovalbumin in BALB/c mice, which were subcutaneously immunized, in terms of antibody production (IgG1, IgG2a, IgG2b, and IgG3), and assessed the levels of IFN-γ in the splenocyte culture using indirect ELISA. Furthermore, we investigated in vitro cytotoxicity of xanthan in the embryo fibroblasts cell line of the NIH/3T3 mouse by MTT assay and propidium iodide uptake assay. The mice immunized with ovalbumin plus xanthan gum exhibited higher antibody IgG1 responses than control groups. Furthermore, the xanthan polysaccharide was capable of increasing the immunogenicity of antigens by producing IFN-γ and did not exhibit cytotoxicity effects in NIH/3T3 mouse fibroblast cells, considered a promising candidate for vaccine adjuvant.

scholarly journals Effect of heat treatment on cytotoxicity of self-adhesive resin cements: Cell viability analysis

2018 ◽  
Vol 12 (02) ◽  
pp. 281-286 ◽  
Author(s):  
Celso Afonso Klein-Júnior ◽  
Roberto Zimmer ◽  
Guilherme Scotta Hentschke ◽  
Denise Cantarelli Machado ◽  
Rubem Beraldo dos Santos ◽  
...  
Keyword(s):  
Heat Treatment ◽  
Cell Viability ◽  
Fibroblast Cell ◽  
Resin Cements ◽  
Adhesive Resin ◽  
Viability Analysis ◽  
3T3 Fibroblasts ◽  
Mixing Control ◽  
Nih 3T3

ABSTRACT Objective: The aim of the study was to assess, in vitro, the influence on cytotoxicity of heat treatment applied before photopolymerization, while mixing three self-adhesive resin cements, in an NIH/3T3 fibroblast cell culture, based on cell viability measures. Methods: Samples were divided into three groups: (1) no heat treatment while mixing (control), (2) 37°C, and (3) 60°C heat treatment while mixing. Cements were light-cured immediately after mixing and immersed in Dulbecco's Modified Eagle Media for the extraction of possibly uncured products after 24 h and 7 days. Cultures contained 0.5 mL of NIH/3T3 fibroblasts per well at a concentration of 0.4 × 105 cells/mL and specific extracts for each sample. Statistical Analysis Used: Data were statistically analyzed with ANOVA and post hoc Student–Newman–Keuls (significance of 5%). Results: Cement cytotoxicity increased with time, as shown by the higher values observed at 7 days. There was a slight difference in intragroup cytotoxicity levels between 24 h and 7 days. Heat treatment at 60°C was associated with a major decrease in cytotoxicity levels in all three groups, both at 24 h and at 7 days, with no differences among the cements. Conclusions: Heat treatment at 60°C should be considered as a strategy to reduce cytotoxicity of self-adhesive resin cements, as evidenced by the results observed at 24 h and 7 days of analysis.

scholarly journals Development of Bionanocomposites Based on PLA, Collagen and AgNPs and Characterization of Their Stability and In Vitro Biocompatibility

2020 ◽  
Vol 10 (7) ◽  
pp. 2265
Author(s):  
Maria Râpă ◽  
Laura Mihaela Stefan ◽  
Traian Zaharescu ◽  
Ana-Maria Seciu ◽  
Anca Andreea Țurcanu ◽  
...  
Keyword(s):  
Normal Growth ◽  
Fibroblast Cell ◽  
Stability Study ◽  
Mouse Fibroblast ◽  
Poly Lactic Acid ◽  
In Vitro Biocompatibility ◽  
Diphenyltetrazolium Bromide ◽  
Mouse Fibroblast Cell Line ◽  
The Stability

Bionanocomposites including poly(lactic acid) (PLA), collagen, and silver nanoparticles (AgNPs) were prepared as biocompatible and stable films. Thermal properties of the PLA-based bionanocomposites indicated an increase in the crystallinity of PLA plasticized due to a small quantity of AgNPs. The results on the stability study indicate the promising contribution of the AgNPs on the durability of PLA-based bionanocomposites. In vitro biocompatibility conducted on the mouse fibroblast cell line NCTC, clone 929, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed high values of cell viability (>80%) after cell cultivation in the presence of bionanocomposite formulations for 48 h, while the percentages of lactate dehydrogenase (LDH) released in the culture medium were reduced (<15%), indicating no damages of the cell membranes. In addition, cell cycle analysis assessed by flow cytometry indicated that all tested bionanocomposites did not affect cell proliferation and maintained the normal growth rate of cells. The obtained results recommend the potential use of PLA-based bionanocomposites for biomedical coatings.

Parasiticidal activity of bovine lactoperoxidase against Toxoplasma gondii

2006 ◽  
Vol 84 (5) ◽  
pp. 774-779 ◽  
Author(s):  
Tetsuya Tanaka ◽  
Shin Murakami ◽  
Haruto Kumura ◽  
Ikuo Igarashi ◽  
Kei-ichi Shimazaki
Keyword(s):  
Toxoplasma Gondii ◽  
Cell Line ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
A Cell ◽  
Mouse Fibroblast Cell Line ◽  
Infected Meat ◽  
Free Environment ◽  
Nih 3T3 ◽  
Mouse Macrophage Cell Line

Toxoplasma gondii is an obligatory intracellular parasitic protozoan transmitted via the ingestion of raw, infected meat that causes congenital infections. In a cell-free environment, virulent Toxoplasma was strikingly resistant to H2O2. The activity of H2O2 or H2O2 generated by glucose – glucose oxidase against the resistant tachyzoite stage of pathogenic T. gondii was enhanced by adding KI and bovine lactoperoxidase (bLPO), referred to here as the bLPO system. Replacing bLPO (heme content, 90%) with recombinant bLPO (heme content, 6%) did not enhance the parasiticidal activity with KI and H2O2. These results indicated that heme contributed to the enzyme activity and resulted in the killing of tachyzoites of T. gondii. Tachyzoites treated with the bLPO system also lost the ability to penetrate the mouse fibroblast cell line (NIH/3T3), and could be killed intracellularly after exposure by bLPO to a mouse macrophage cell line (J774A.1). These findings suggested that toxicity was mediated through small amounts of H2O2 generated by phagocytic events in naive macrophages, and by the peroxidative activity of bLPO. Our observations suggest that the bLPO system could help prevent the development of Toxoplasmosis in humans after ingesting raw, infected meat.

Comparison of Validity between WST-1 and MTT Test in Bioceramic Materials

2005 ◽  
Vol 284-286 ◽  
pp. 585-588 ◽  
Author(s):  
Kwang Mahn Kim ◽  
Sang Bae Lee ◽  
Se Ho Lee ◽  
Yong Keun Lee ◽  
Kyoung Nam Kim
Keyword(s):  
Clinical Evaluation ◽  
Water Solubility ◽  
Fibroblast Cell ◽  
Storage Condition ◽  
Mouse Fibroblast ◽  
Cytotoxicity Test ◽  
Mtt Test ◽  
In Vitro Biocompatibility ◽  
Proliferation Assays

Cytotoxicity test was essential for the pre-clinical evaluation of bioceramics. Proliferation assays such as MTT, XTT and WST-1 were commonly used for measuring biocompatibility. WST-1 was more convenient than MTT because of its water-solubility and storage condition. The calcium phosphate glass and β-TCP have been used for bone substitute, and some magnetic ferrites have been used for hyperthermic treatment. L929, mouse fibroblast cell, was the representative cell-line for in vitro biocompatibility test. The extracts of test samples were prepared by ISO10993-12:2002. The biocompatibilities of the extracts were measured by MTT and WST-1 assay and their pH were measured with pH meter. The cellular survival rate of CPG was the lowest and the results of the WST-1 test showed results similar to those of the MTT test. Thus, proliferation assays using WST-1 may be conveniently and routinely applicable to pre-clinical evaluation of bioceramics.

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