Improving the interferences of methyl thiazolyl tetrazolium and IL-8 assays in assessing the cytotoxicity of nanoparticles

I. Lun Hsiao, Yuh Jeen Huang

研究成果: 雜誌貢獻文章

10 引文 (Scopus)

摘要

Methyl thiazolyl tetrazolium (MTT) and interleukin-8 (IL-8) assays are common colorimetric methods to measure mitochondrial activity and drug induced pro-inflammatory factors. However, many reports have described how MTT absorbance and cytokine adsorption could limit their applicability in evaluating the cytotoxicity of nanomaterials. In this study, we used an acid-containing isopropanol complex as a substitute for dimethyl sulfoxide (DMSO) solvent to dissolve MTT formazan, which was expected to diminish the absorbance of nano-ZnO at 570 nm where maximum absorbance for the MTT formazan was detected. In addition, we used a serum-containing medium to prevent the possible effects of IL-8 protein adsorption in the nano-ZnO and nano-TiO 2. The results showed that the modified method by using acid-containing isopropanol step in MTT assay, nano-ZnO exposed to human lung epithelial cells had the lowest cell viability (from 12.5 to 50 μg mL -1) and EC50 value (8.4 μg mL -1) comparing with the conventional MTT protocol or adding phosphate buffered saline (PBS) to wash cells. The reason for this was the acid-containing isopropanol completely dissolved nano-ZnO with no additional absorbance when compared to the background solvent at 570 nm. On the other hand, the IL-8 protein had a marked influence on the adsorption of nano-TiO 2 in the serum-free medium. While only at 100 μg mL-1 of nano-ZnO, an influence on the adsorption of IL-8 was observed. This could be attributed to the different charges on the surface of nanomaterials. This problem could be overcome through the addition of fetal bovine serum (FBS) to the medium.
原文英語
頁(從 - 到)5228-5233
頁數6
期刊Journal of Nanoscience and Nanotechnology
11
發行號6
DOIs
出版狀態已發佈 - 2011
對外發佈Yes

指紋

interleukins
Cytotoxicity
Interleukin-8
Nanoparticles
Adsorption
2-Propanol
Assays
serums
Formazans
interference
nanoparticles
adsorption
Nanostructures
Nanostructured materials
acids
Acids
proteins
Proteins
Dimethyl sulfoxide
Serum-Free Culture Media

ASJC Scopus subject areas

  • Bioengineering
  • Chemistry(all)
  • Biomedical Engineering
  • Materials Science(all)
  • Condensed Matter Physics

引用此文

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title = "Improving the interferences of methyl thiazolyl tetrazolium and IL-8 assays in assessing the cytotoxicity of nanoparticles",
abstract = "Methyl thiazolyl tetrazolium (MTT) and interleukin-8 (IL-8) assays are common colorimetric methods to measure mitochondrial activity and drug induced pro-inflammatory factors. However, many reports have described how MTT absorbance and cytokine adsorption could limit their applicability in evaluating the cytotoxicity of nanomaterials. In this study, we used an acid-containing isopropanol complex as a substitute for dimethyl sulfoxide (DMSO) solvent to dissolve MTT formazan, which was expected to diminish the absorbance of nano-ZnO at 570 nm where maximum absorbance for the MTT formazan was detected. In addition, we used a serum-containing medium to prevent the possible effects of IL-8 protein adsorption in the nano-ZnO and nano-TiO 2. The results showed that the modified method by using acid-containing isopropanol step in MTT assay, nano-ZnO exposed to human lung epithelial cells had the lowest cell viability (from 12.5 to 50 μg mL -1) and EC50 value (8.4 μg mL -1) comparing with the conventional MTT protocol or adding phosphate buffered saline (PBS) to wash cells. The reason for this was the acid-containing isopropanol completely dissolved nano-ZnO with no additional absorbance when compared to the background solvent at 570 nm. On the other hand, the IL-8 protein had a marked influence on the adsorption of nano-TiO 2 in the serum-free medium. While only at 100 μg mL-1 of nano-ZnO, an influence on the adsorption of IL-8 was observed. This could be attributed to the different charges on the surface of nanomaterials. This problem could be overcome through the addition of fetal bovine serum (FBS) to the medium.",
keywords = "A549 cells, Absorption effect, IL-8, MTT, Protein adsorption",
author = "Hsiao, {I. Lun} and Huang, {Yuh Jeen}",
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journal = "Journal of Nanoscience and Nanotechnology",
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T1 - Improving the interferences of methyl thiazolyl tetrazolium and IL-8 assays in assessing the cytotoxicity of nanoparticles

AU - Hsiao, I. Lun

AU - Huang, Yuh Jeen

PY - 2011

Y1 - 2011

N2 - Methyl thiazolyl tetrazolium (MTT) and interleukin-8 (IL-8) assays are common colorimetric methods to measure mitochondrial activity and drug induced pro-inflammatory factors. However, many reports have described how MTT absorbance and cytokine adsorption could limit their applicability in evaluating the cytotoxicity of nanomaterials. In this study, we used an acid-containing isopropanol complex as a substitute for dimethyl sulfoxide (DMSO) solvent to dissolve MTT formazan, which was expected to diminish the absorbance of nano-ZnO at 570 nm where maximum absorbance for the MTT formazan was detected. In addition, we used a serum-containing medium to prevent the possible effects of IL-8 protein adsorption in the nano-ZnO and nano-TiO 2. The results showed that the modified method by using acid-containing isopropanol step in MTT assay, nano-ZnO exposed to human lung epithelial cells had the lowest cell viability (from 12.5 to 50 μg mL -1) and EC50 value (8.4 μg mL -1) comparing with the conventional MTT protocol or adding phosphate buffered saline (PBS) to wash cells. The reason for this was the acid-containing isopropanol completely dissolved nano-ZnO with no additional absorbance when compared to the background solvent at 570 nm. On the other hand, the IL-8 protein had a marked influence on the adsorption of nano-TiO 2 in the serum-free medium. While only at 100 μg mL-1 of nano-ZnO, an influence on the adsorption of IL-8 was observed. This could be attributed to the different charges on the surface of nanomaterials. This problem could be overcome through the addition of fetal bovine serum (FBS) to the medium.

AB - Methyl thiazolyl tetrazolium (MTT) and interleukin-8 (IL-8) assays are common colorimetric methods to measure mitochondrial activity and drug induced pro-inflammatory factors. However, many reports have described how MTT absorbance and cytokine adsorption could limit their applicability in evaluating the cytotoxicity of nanomaterials. In this study, we used an acid-containing isopropanol complex as a substitute for dimethyl sulfoxide (DMSO) solvent to dissolve MTT formazan, which was expected to diminish the absorbance of nano-ZnO at 570 nm where maximum absorbance for the MTT formazan was detected. In addition, we used a serum-containing medium to prevent the possible effects of IL-8 protein adsorption in the nano-ZnO and nano-TiO 2. The results showed that the modified method by using acid-containing isopropanol step in MTT assay, nano-ZnO exposed to human lung epithelial cells had the lowest cell viability (from 12.5 to 50 μg mL -1) and EC50 value (8.4 μg mL -1) comparing with the conventional MTT protocol or adding phosphate buffered saline (PBS) to wash cells. The reason for this was the acid-containing isopropanol completely dissolved nano-ZnO with no additional absorbance when compared to the background solvent at 570 nm. On the other hand, the IL-8 protein had a marked influence on the adsorption of nano-TiO 2 in the serum-free medium. While only at 100 μg mL-1 of nano-ZnO, an influence on the adsorption of IL-8 was observed. This could be attributed to the different charges on the surface of nanomaterials. This problem could be overcome through the addition of fetal bovine serum (FBS) to the medium.

KW - A549 cells

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KW - Protein adsorption

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