Evaluation of eight in vitro assays for assessing the cytotoxicity of cigarette smoke condensate

Putnam, Kelly; Bombick, David W.; Doolittle, David J.

doi:10.1016/s0887-2333(02)00050-4

Cited by 161 publications

(117 citation statements)

References 14 publications

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“…They have concluded that assays that measure membrane integrity (LDH) are useful for short exposure times (1 h), NRU assay was the most sensitive for moderate (3-6 h) exposure times; and assays that measure total cell number (NRU and kenacid blue) were more sensitive for longer exposure times (12, 18 and 24 h). 32 But in our study, both phenolics showed similar cytotoxicity profile in NRU and MTT assays in all exposure times. …”

Section: Discussionsupporting

confidence: 55%

“…The NRU and the MTT assays were found to be the most sensitive in detecting cytotoxic events. Putnam et al 32 have also studied cytotoxicity of cigarette smoke condensate with eight different (NRU, LDH release, kenacid blue binding, MTT, XTT, acid phosphatase activity, sulforhodamine B binding and resazurin binding) cytotoxicity assays. Four of the more widely used cytotoxicity assays (NRU, MTT, kenacid blue and LDH) were also evaluated at 3, 6, 12 and 18 h time points in this study.…”

Section: Discussionmentioning

confidence: 99%

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Assessment of Cytotoxicity Profiles of Different Phytochemicals: Comparison of Neutral Red and MTT Assays in Different Cells in Different Time Periods

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et al. 2017

tjps

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ÖZAmaç: Fenolik bileşikler sağlığı koruyucu farklı özellikler gösterir. Galangin, kurkumin, piknogenol, puerarin ve ursolik asit halk tıbbında yaygın olarak kullanılan bitkisel fenoliklerdendir. Bu çalışmanın amacı, nötral kırmızı alım (NKA) ve MTT yöntemleri arasındaki farkı sağlıklı hücreler ve kanser hücrelerinde farklı bitkisel fenoliklerin (galangin, kurkumin, piknogenol, puerarin ve ursolik asit) farklı zaman aralıklarında farklı zaman aralıklarında belirlemektir. Gereç ve Yöntemler: Bu çalışmada, bu fenolik bileşiklerin sitotoksik etkileri sağlıklı hücreler (Çin hamster fibroblast hücre hattı, V79) ve kanser [insan serviks epitelyal adenokarsinoma hücre hattı, Henrietta Lacks (HeLa) ve insan meme karsinoma hücre hattı (BT-474)] hücrelerinde 18, 24 ve 48 saatlik inkübasyon sürelerinde NKA ve MTT yöntemleriyle değerlendirilmiştir. Bulgular: Bulgularımız galangin, kurkumin, piknogenol, puerarin ve ursolik asitin V79, HeLa ve BT-474 hücre canlılıklarını 18, 24 ve 48 saatlik inkübasyon sürelerinde doza bağımlı olarak azalttığını göstermiştir. Ancak en az hücre canlılık oranı 48 saatlik inkübasyon sonrası görülmüştür. NKA ile MTT yöntemlerinin sonuçları arasında fark görülmemiştir. Sonuç: Sitotoksisite analizinde kullanılacak yöntem ve inkübasyon süresinin belirlenmesi için maddelerin sitotoksisite mekanizması bilinmelidir. Anahtar kelimeler: MTT, nötral kırmızı, bitkisel fenoliklerObjectives: Phenolic compounds exhibit several health protective properties. Galangin, curcumin, pycnogenol, puerarin and ursolic acid are commonly used plant phenolics in folk medicine. The aim of our study was to evaluate the difference between neutral red uptake (NRU) and MTT assays using different plant phenolics (galangin, curcumin, pycnogenol, puerarin and ursolic acid) in healthy and cancer cells in different time periods. Materials and Methods: In this study, the cytotoxic effects of these phenolic compounds were investigated by NRU and MTT assays in healthy (V79, Chinese hamster fibroblast cell line) and cancer [human cervix epithelial adenocarcinoma cell line Henrietta Lacks (HeLa) and human mammary carcinoma cell line (BT-474)] in 18, 24 and 48 h incubation periods. Results: Our results demonstrated that galangin, curcumin, pycnogenol, puerarin and ursolic acid decreased cell viability of V79, HeLa and BT-474 cells in a dose-dependent manner in 18, 24 and 48 h incubation periods. However, the cell survival rate was much lower in 48 h incubation period. There was no difference between the results from NRU and MTT assays. Conclusion: To decide which incubation period and which cytotoxicity study to be used, the cytotoxicity mechanism of the compound must be known.

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Section: Discussionsupporting

confidence: 55%

Section: Discussionmentioning

confidence: 99%

Assessment of Cytotoxicity Profiles of Different Phytochemicals: Comparison of Neutral Red and MTT Assays in Different Cells in Different Time Periods

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Anlara

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et al. 2017

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“…Interestingly, CSE is expected to have dose-dependent effect on elastin synthesis and deposit. However, in our experiment, this effect was not observed, which was consistent with other findings (Chen et al, 2005;Putnam et al, 2002). Cigarette smoke was a complex compound, different doses of CSE might inspire different intracellular signal pathways (Messner et al, 2012).…”

Section: Cigarette Causes Poor Proliferation Ability Viability and Esupporting

confidence: 92%

Knockdown of versican 1 blocks cigarette-induced loss of insoluble elastin in human lung fibroblasts

Xu¹,

Lu²,

Zhang³

et al. 2015

Respiratory Physiology & Neurobiology

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of versican 1 blocks cigarette-induced loss of insoluble elastin in human lung fibroblasts, Respiratory Physiology and Neurobiology (2015), http://dx.doi.org/10.1016/j.resp.2015. 05.004 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. HighlightsWe find extracellular matrix protein versican 1 is in creased by cigarette smoke extract solution in lung fibroblasts.Versican 1 knockdown restore cigarette-related elastin loss through chondroitin sulfate decrease.Exgenous chondroitin sulfate could block the beneficent effect brought by versican 1 knockdown. Page 2 of 21A c c e p t e d M a n u s c r i p t Abstract COPD lung is characterized by loss of alveolar elastic fibers and an increase in the chondroitin sulfate (CS) matrix proteoglycan versican 1 (V1). V1 is a known inhibitor of elastic fiber deposition and this study investigates the effects of knockdown of V1, and add-back of CS, on CCL-210 lung fibroblasts treated with cigarette smoke extract (CSE) as a model for COPD. CSE inhibited fibroblast proliferation, viability, tropoelastin synthesis, and elastin deposition, and increased V1 synthesis and secretion. V1 siRNA decreased V1 and constituent CS, did not affect tropoelastin production, but blocked the CSE-induced loss in insoluble elastin. Exogenous CS reduced insoluble elastin, even in the presence of V1 siRNA. These findings confirm that V1 and CS impair the assembly of tropoelastin monomers into insoluble fibers, and further demonstrate that specific knockdown of V1 alleviates the impaired assembly of elastin seen in cultures of pulmonary fibroblasts exposed to CSE, indicating a regulatory role for this protein in the pathophysiology of COPD.

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“…Neutral red is incorporated into lysosomes of living cells, and therefore an indicator of the proportion of viable cells with an active endocytosis process. The chemicals that damage membranes will decrease the cell's ability to take up neutral red [19]. Our in vitro cytotoxicity tests clearly indicated that the HCM and KMC-C extracts acted as cytotoxic agents and caused significant reduction of cell metabolic activity and cell viability.…”

Section: +mentioning

confidence: 70%

Do novel cement-type biomaterials reveal ion reactivity that affects cell viability in vitro?

et al. 2013

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Calcium phosphate bioceramics have been studied as bone filler materials for years and have become a component of many commercial products. It is widely known that surface-reactive biomaterials may cause changes in the concentration of crucial ions in the surrounding environment, thereby affecting cell metabolism and viability. The aim of this study was to produce five cement-type biomaterials and characterize their phase composition using X-ray diffraction method, and porosity and pore size distribution using mercury intrusion porosimeter. We then evaluated ion interactions of the novel biomaterials with the surrounding environment (culture medium). A commercially available bone substitute, HydroSet™ (Stryker®), was used as a reference. MTT and NRU cytotoxicity tests were performed to assess the effect of changes in the concentration of crucial ions (calcium, magnesium, phosphate) on osteoblast metabolism and viability in vitro. Our study clearly indicated that various biomaterials demonstrated different ion reactivity and consequently may cause changes in ion concentration in the local environment. Critically low or high values of calcium, magnesium, and phosphate concentrations in the medium exerted cytotoxic effects on the cultured cells. Moreover, we discovered that the chemical composition of the culture medium had a substantial influence on ion interactions with biomaterials.

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Evaluation of eight in vitro assays for assessing the cytotoxicity of cigarette smoke condensate

Cited by 161 publications

References 14 publications

Assessment of Cytotoxicity Profiles of Different Phytochemicals: Comparison of Neutral Red and MTT Assays in Different Cells in Different Time Periods

Assessment of Cytotoxicity Profiles of Different Phytochemicals: Comparison of Neutral Red and MTT Assays in Different Cells in Different Time Periods

Knockdown of versican 1 blocks cigarette-induced loss of insoluble elastin in human lung fibroblasts

Do novel cement-type biomaterials reveal ion reactivity that affects cell viability in vitro?

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