scholarly journals A Combined Assay of Cell Viability and in Vitro Cytotoxicity with a Highly Water-Soluble Tetrazolium Salt, Neutral Red and Crystal Violet.

1996 ◽  
Vol 19 (11) ◽  
pp. 1518-1520 ◽  
Author(s):  
Munetaka ISHIYAMA ◽  
Hideyuki TOMINAGA ◽  
Masanobu SHIGA ◽  
Kazumi SASAMOTO ◽  
Yosuke OHKURA ◽  
...  
Keyword(s):  
Cell Viability ◽  
Crystal Violet ◽  
Neutral Red ◽  
In Vitro Cytotoxicity ◽  
Water Soluble ◽  
Tetrazolium Salt

scholarly journals Screening of different cytotoxicity methods for the assessment of ENDS toxicity relative tobacco cigarettes

2021 ◽  
Author(s):  
Massimo Caruso ◽  
Rosalia Emma ◽  
Sonja Rust ◽  
Alfio Distefano ◽  
Giuseppe Carota ◽  
...  
Keyword(s):  
Cell Viability ◽  
Electronic Cigarettes ◽  
Annexin V ◽  
Initial Step ◽  
Neutral Red ◽  
In Vitro Cytotoxicity ◽  
The European Union ◽  
Time Resolved ◽  
Cytotoxicity Studies

AbstractElectronic Nicotine Delivery Systems (ENDS), i.e., electronic cigarettes (e-cigs) and Tobacco Heating Products (THPs), are rapidly growing in popularity. The marketing of these products is regulated by specific rules in the European Union and in the US, which permit their legal sales. Nonetheless, comprehensive quality and safety requirements for regulatory purposes are still under development. Cytotoxicity studies are an important initial step in appraising the potential toxicity of ENDS. The aim of the present study was to screen a battery of different in vitro cytotoxicity methods for the assessment of toxicity induced by ENDS. We evaluated different cytotoxicity assays, including neutral red uptake (NRU), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Annexin V apoptosis, High Content Screening (HCS) assays and Real Time Cell Analysis (RTCA), to compare two e-cigs (Vype ePen 3 and Vype eStick Maxx) and two THPs (IQOS and GLO™) with the 1R6F reference tobacco cigarette. Human bronchial epithelial cells (H292) were exposed to 1R6F smoke (5 puffs by HCI regime), ePen vapor (10 puffs by modified HCI regime), eStick vapor (25 puffs by CRM81 regime), IQOS vapor (7 puffs by HCI regime) and GLO vapor (8 puffs by HCI regime) at air-liquid interface. All tests showed reduced cell viability following 1R6F smoke exposure and slight or no reduction with ENDS at 24 hours compared to controls. In addition, Annexin V and RTCA exhibited a further significant reduction in cell viability following 1R6F exposure compared with other assays. Furthermore, Annexin V allowed to discriminate viable cells from those in early/late apoptosis. Finally, RTCA and HCS being time-resolved analyses allowed also to determine the kinetic dependency parameter for toxicity of smoke/vapor chemicals on cell viability. In conclusion, NRU assay may be considered a suitable test, especially when combined with a time-resolved test, for assessing the kinetic of cytotoxicity induced by these products.

Cytotoxicity of MEIC Chemicals to Rainbow Trout R1 Cell Line and Multivariate Comparison with Ecotoxicity Tests

1997 ◽  
Vol 25 (3) ◽  
pp. 331-338
Author(s):  
Helmut Segner ◽  
Gerrit Schüürmann
Keyword(s):  
Rainbow Trout ◽  
Cell Line ◽  
Crystal Violet ◽  
Mammalian Cells ◽  
Cell Attachment ◽  
Neutral Red ◽  
Test System ◽  
In Vitro Cytotoxicity ◽  
Ecotoxicity Tests

The cytotoxic effects of the first ten chemicals from the Multicenter Evaluation of In Vitro Cytotoxicity (MEIC) programme on the fibroblast-like R1 cell line, derived from rainbow trout (Oncorhynchus mykiss), were measured by using three endpoints: cell attachment (24-hour exposure, crystal violet protein stain); cell viability (24-hour exposure, neutral red uptake cytotoxicity assay); and cell growth (144-hour exposure, crystal violet protein stain). The results were compared with published MEIC toxicity data from fish and mammalian cells, and from ecotoxicity tests with bacteria, invertebrates and plants by using multivariate statistical techniques. For eight of the ten compounds under consideration, a high degree of correlation in toxicity ranking between the various bioassays was observed, irrespective of the test systems or endpoints utilised. This similarity might be explained by the basal cytotoxicity concept. The 20% dissimilarity of the results, however, indicates the influence of test system-specific or end-point-specific sensitivities.

Formulation, Characterization and In vitro Cytotoxicity of 5-Fluorouracil Loaded Polymeric Electrospun Nanofibers for the Treatment of Skin Cancer

2019 ◽  
Vol 13 (2) ◽  
pp. 114-128 ◽  
Author(s):  
Gayatri Patel ◽  
Bindu K.N. Yadav
Keyword(s):  
Skin Cancer ◽  
Basal Cell Carcinoma ◽  
Cell Viability ◽  
Cell Carcinoma ◽  
Basal Cell ◽  
Fiber Diameter ◽  
Electrospun Nanofibers ◽  
In Vitro Cytotoxicity ◽  
Fractional Factorial

Background: The purpose of this study was to formulate, characterize and conduct in vitro cytotoxicity of 5-fluorouracil loaded polymeric electrospun nanofibers for the treatment of skin cancer. The patents on electrospun nanofibers (US9393216B2), (US14146252), (WO2015003155A1) etc. helped in the selection of polymers and method for the preparation of nanofibers. Methods: In the present study, the fabrication of nanofibers was done using a blend of chitosan with polyvinyl alcohol and processed using the electrospinning technique. 5-fluorouracil with known chemotherapeutic potential in the treatment of skin cancer was used as a drug carrier. 24-1 fractional factorial screening design was employed to study the effect of independent variables like the concentration of the polymeric solution, applied voltage (kV), distance (cm), flow rate (ml / hr) on dependent variables like % entrapment efficiency and fiber diameter. Results: Scanning electron microscopy was used to characterize fiber diameter and morphology. Results showed that the fiber diameter of all batches was found in the range of 100-200 nm. The optimized batch results showed the fiber diameter of 162.7 nm with uniform fibers. The tensile strength obtained was 190±37 Mpa. Further in vitro and ex vivo drug release profile suggested a controlled release mechanism for an extended period of 24 hr. The 5-fluorouracil loaded electrospun nanofibers were found to decrease cell viability up to ≥50% over 24 hr, with the number of cells dropping by ~ 10% over 48 hr. As the cell viability was affected by the release of 5-fluorouracil, we believe that electrospun nanofibers are a promising drug delivery system for the treatment of Basal Cell Carcinoma (BCC) skin cancer. Conclusion: These results demonstrate the possibility of delivering 5-Fluorouracil loaded electrospun nanofiber to skin with enhanced encapsulation efficiency indicating the effectiveness of the formulation for the treatment of basal cell carcinoma type of skin cancer.

The Development and Evaluation of In Vitro Tests by the FRAME Alternatives Laboratory

1995 ◽  
Vol 23 (1) ◽  
pp. 75-90
Author(s):  
Richard H. Clothier ◽  
Karen A. Atkinson ◽  
Michael J. Garle ◽  
Rachel K. Ward ◽  
Angela Willshaw
Keyword(s):  
Research Programme ◽  
Neutral Red ◽  
In Vitro Cytotoxicity ◽  
In Vitro Tests ◽  
Dermal Toxicity ◽  
Mechanisms Of Toxicity ◽  
The Us ◽  
Detergent Association ◽  
Assay Methods

This review outlines the work which has been conducted in the FRAME Alternatives Laboratory during the first ten years of the FRAME Research Programme. A number of in vitro tests, including the kenacid blue, neutral red release and fluorescein leakage assay methods, have been evaluated and have subsequently been included in validation schemes organised by the US Soap and Detergent Association, the US Cosmetic, Toiletry and Fragrance Association, the European Commission and the European Cosmetic, Toiletry and Perfumery Association, as well as in the Scandinavian multicentre evaluation of in vitro cytotoxicity testing scheme. More recently, research has been undertaken in the areas of phototoxicity, immunotoxicity, dermal toxicity and intercellular communication, in addition to investigations into fundamental mechanisms of toxicity.

scholarly journals Comparative Study of Cigarette Smoke Cytotoxicity Using Two In Vitro Assay Systems

2014 ◽  
Vol 26 (3) ◽  
pp. 98-108
Author(s):  
Toshiro Fukushima ◽  
Hitomi Tanaka ◽  
Takeshi Yamamoto
Keyword(s):  
Cigarette Smoke ◽  
Rank Order ◽  
In Vitro Cytotoxicity ◽  
Water Soluble ◽  
Cytotoxic Agents ◽  
Main Stream ◽  
Total Particulate Matter ◽  
Vitro Assay ◽  
Modes Of Action

SUMMARYThe aim of this study was to compare the results obtained from two in vitro cytotoxicity assays that depend upon different mechanisms/modes of action. The Neutral Red Uptake (NRU) assay is based on endocytotic activity whereas the Water Soluble Tetrazolium Salts (WST-1) assay is based on mitochondrial dehydrogenase activity. Both were investigated in light of their wide use and documented validation. The total particulate matter (TPM) and gas vapor phase (GVP) of main stream smoke derived from Kentucky reference cigarettes 3R4F and 10 test cigarettes made of 100% flue-cured or 100% Burley tobacco were individually applied to the two assays using CHO-K1 cells. In addition, cigarette smoke constituents and known cytotoxic agents, documented to affect specific endpoints, were evaluated within both assays. Although the NRU assay was primarily more sensitive than the WST-1 assay, both assays provided comparable results in terms of the rank order for the cytotoxicity of cigarette smoke samples. In addressing the cytotoxicity of constituents in cigarette smoke, acrolein, hydroquinone and catechol gave clear dose-related decreases in cell viability (an end point common in both assays). Moreover, enzyme inhibitors of the mitochondrial respiratory chain and chemicals causing membrane disruption also showed similar responses regardless of the specific endpoint addressed within the cytotoxicity assay. In conclusion, results from the NRU and WST-1 assay are comparable therefore indicating results were independent of the different assay detection mechanisms/modes of action. [Beitr. Tabakforsch. Int. 26 (2014) 98-108]

scholarly journals Multiresponsive Hybrid Microparticles for Stimuli-Responsive Delivery of Bioactive Compounds

2020 ◽  
Vol 10 (12) ◽  
pp. 4324 ◽  
Author(s):  
Sergei S. Vlasov ◽  
Pavel S. Postnikov ◽  
Mikhail V. Belousov ◽  
Sergei V. Krivoshchekov ◽  
Mekhman S. Yusubov ◽  
...  
Keyword(s):  
Cell Viability ◽  
Spherical Shape ◽  
In Vitro Cytotoxicity ◽  
Poly Lactic Acid ◽  
Iron Core ◽  
Stimuli Responsive ◽  
Ultrasonic Fields ◽  
Burst Intensity ◽  
Free Doxorubicin

Hybrid microparticles based on an iron core and an amphiphilic polymeric shell have been prepared to respond simultaneously to magnetic and ultrasonic fields and variation in the surrounding pH to trigger and modulate the delivery of doxorubicin. The microparticles have been developed in four steps: (i) synthesis of the iron core; (ii) surface modification of the core; (iii) conjugation with the amphiphilic poly(lactic acid)-grafted chitosan; and (iv) doxorubicin loading. The particles demonstrate spherical shape, a size in the range of 1–3 µm and surface charge that is tuneable by changing the pH of the environment. The microparticles demonstrate good stability in simulated physiological solutions and are able to hold up to 400 µg of doxorubicin per mg of dried particles. The response to ultrasound and the changes in the shell structure during exposure to different pH levels allows the control of the burst intensity and release rate of the payload. Additionally, the magnetic response of the iron core is preserved despite the polymer coat. In vitro cytotoxicity tests performed on fibroblast NIH/3T3 demonstrate a reduction in the cell viability after administration of doxorubicin-loaded microparticles compared to the administration of free doxorubicin. The application of ultrasound causes a burst in the release of the doxorubicin from the carrier, causing a decrease in cell viability. The microparticles demonstrate in vitro cytocompatibility and hemocompatibility at concentrations of up to 50 and 60 µg/mL, respectively.

scholarly journals Preparation, Cytotoxicity, and In Vitro Bioimaging of Water Soluble and Highly Fluorescent Palladium Nanoclusters

2020 ◽  
Vol 7 (1) ◽  
pp. 20 ◽  
Author(s):  
Suresh Thangudu ◽  
Poliraju Kalluru ◽  
Raviraj Vankayala
Keyword(s):  
In Vitro Cytotoxicity ◽  
Molecular Probes ◽  
Water Soluble ◽  
Quantum Yields ◽  
Cell Labelling ◽  
Metal Nanoclusters ◽  
Extinction Coefficients ◽  
Cytotoxicity Studies

Fluorescent probes offer great potential to identify and treat surgical tumors by clinicians. To this end, several molecular probes were examined as in vitro and in vivo bioimaging probes. However, due to their ultra-low extinction coefficients as well as photobleaching problems, conventional molecular probes limit its practical utility. To address the above mentioned challenges, metal nanoclusters (MNCs) can serve as an excellent alternative with many unique features such as higher molar extinction coefficients/light absorbing capabilities, good photostability and appreciable fluorescence quantum yields. Herein, we reported a green synthesis of water soluble palladium nanoclusters (Pd NCs) and characterized them by using various spectroscopic and microscopic characterization techniques. These nanoclusters showed excellent photophysical properties with the characteristic emission peak centered at 500 nm under 420 nm photoexcitation wavelength. In vitro cytotoxicity studies in human cervical cancer cells (HeLa) cells reveal that Pd NCs exhibited good biocompatibility with an IC50 value of >100 µg/mL and also showed excellent co-localization and distribution throughout the cytoplasm region with a significant fraction translocating into cell nucleus. We foresee that Pd NCs will carry huge potential to serve as a new generation bioimaging nanoprobe owing to its smaller size, minimal cytotoxicity, nucleus translocation capability and good cell labelling properties.

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