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Iron nanoparticles increase 7-ketocholesterol-induced cell death, inflammation, and oxidation on murine cardiac HL1-NB cells.

Kahn E, Baarine M, Pelloux S, Riedinger JM, Frouin F, Tourneur Y, Lizard G - Int J Nanomedicine (2010)

Bottom Line: Pro-inflammatory effects are estimated by ELISA in order to quantify IL-8 and MCP-1 secretions.Pro-oxidative effects are measured with hydroethydine (HE).They induce a slight LDH release, and have no inflammatory or oxidative effects.

View Article: PubMed Central - PubMed

Affiliation: INSERM U678/UMR - S UPMC, IFR 14, CHU Pitié-Salpêtrière, 75634 Paris Cedex 13, France. kahn@imed.jussieu.fr

ABSTRACT

Objective: To evaluate the cytotoxicity of iron nanoparticles on cardiac cells and to determine whether they can modulate the biological activity of 7-ketocholesterol (7KC) involved in the development of cardiovascular diseases. Nanoparticles of iron labeled with Texas Red are introduced in cultures of nonbeating mouse cardiac cells (HL1-NB) with or without 7-ketocholesterol 7KC, and their ability to induce cell death, pro-inflammatory and oxidative effects are analyzed simultaneously.

Study design: Flow cytometry (FCM), confocal laser scanning microscopy (CLSM), and subsequent factor analysis image processing (FAMIS) are used to characterize the action of iron nanoparticles and to define their cytotoxicity which is evaluated by enhanced permeability to SYTOX Green, and release of lactate deshydrogenase (LDH). Pro-inflammatory effects are estimated by ELISA in order to quantify IL-8 and MCP-1 secretions. Pro-oxidative effects are measured with hydroethydine (HE).

Results: Iron Texas Red nanoparticles accumulate at the cytoplasmic membrane level. They induce a slight LDH release, and have no inflammatory or oxidative effects. However, they enhance the cytotoxic, pro-inflammatory and oxidative effects of 7KC. The accumulation dynamics of SYTOX Green in cells is measured by CLSM to characterize the toxicity of nanoparticles. The emission spectra of SYTOX Green and nanoparticles are differentiated, and corresponding factor images specify the possible capture and cellular localization of nanoparticles in cells.

Conclusion: The designed protocol makes it possible to show how Iron Texas Red nanoparticles are captured by cardiomyocytes. Interestingly, whereas these fluorescent iron nanoparticles have no cytotoxic, pro-inflammatory or oxidative activities, they enhance the side effects of 7KC.

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Related in: MedlinePlus

Basic presentation of FAMIS. Factors are estimated in a two-step procedure from the image sequence: 1) correspondence analysis and 2) oblique analysis are performed to obtain positive factor curves and images. Factor images are recomputed back to the original sampling by oblique projection on the factor curves.
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f1-ijn-5-185: Basic presentation of FAMIS. Factors are estimated in a two-step procedure from the image sequence: 1) correspondence analysis and 2) oblique analysis are performed to obtain positive factor curves and images. Factor images are recomputed back to the original sampling by oblique projection on the factor curves.

Mentions: Sequences of images were further processed by FAMIS, available at Apteryx under the name of Pixies (www.apteryx.fr). FAMIS decomposes image sequences into a smaller number of images called factor images, and curves, called factor curves.35,36 Factor curves estimate individual spectral or temporal behavior in the sequence of images. Factor images correspond to spatial distribution components. The basic idea of FAMIS is to process the curves that represent the evolution of fluorescence intensity of each pixel in the image spectral or temporal sequence (Figure 1). FAMIS assumes that each pixel is a mixture of different patterns and aims to unmix them. Factors are estimated in a two-step procedure from the image sequence.


Iron nanoparticles increase 7-ketocholesterol-induced cell death, inflammation, and oxidation on murine cardiac HL1-NB cells.

Kahn E, Baarine M, Pelloux S, Riedinger JM, Frouin F, Tourneur Y, Lizard G - Int J Nanomedicine (2010)

Basic presentation of FAMIS. Factors are estimated in a two-step procedure from the image sequence: 1) correspondence analysis and 2) oblique analysis are performed to obtain positive factor curves and images. Factor images are recomputed back to the original sampling by oblique projection on the factor curves.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2865013&req=5

f1-ijn-5-185: Basic presentation of FAMIS. Factors are estimated in a two-step procedure from the image sequence: 1) correspondence analysis and 2) oblique analysis are performed to obtain positive factor curves and images. Factor images are recomputed back to the original sampling by oblique projection on the factor curves.
Mentions: Sequences of images were further processed by FAMIS, available at Apteryx under the name of Pixies (www.apteryx.fr). FAMIS decomposes image sequences into a smaller number of images called factor images, and curves, called factor curves.35,36 Factor curves estimate individual spectral or temporal behavior in the sequence of images. Factor images correspond to spatial distribution components. The basic idea of FAMIS is to process the curves that represent the evolution of fluorescence intensity of each pixel in the image spectral or temporal sequence (Figure 1). FAMIS assumes that each pixel is a mixture of different patterns and aims to unmix them. Factors are estimated in a two-step procedure from the image sequence.

Bottom Line: Pro-inflammatory effects are estimated by ELISA in order to quantify IL-8 and MCP-1 secretions.Pro-oxidative effects are measured with hydroethydine (HE).They induce a slight LDH release, and have no inflammatory or oxidative effects.

View Article: PubMed Central - PubMed

Affiliation: INSERM U678/UMR - S UPMC, IFR 14, CHU Pitié-Salpêtrière, 75634 Paris Cedex 13, France. kahn@imed.jussieu.fr

ABSTRACT

Objective: To evaluate the cytotoxicity of iron nanoparticles on cardiac cells and to determine whether they can modulate the biological activity of 7-ketocholesterol (7KC) involved in the development of cardiovascular diseases. Nanoparticles of iron labeled with Texas Red are introduced in cultures of nonbeating mouse cardiac cells (HL1-NB) with or without 7-ketocholesterol 7KC, and their ability to induce cell death, pro-inflammatory and oxidative effects are analyzed simultaneously.

Study design: Flow cytometry (FCM), confocal laser scanning microscopy (CLSM), and subsequent factor analysis image processing (FAMIS) are used to characterize the action of iron nanoparticles and to define their cytotoxicity which is evaluated by enhanced permeability to SYTOX Green, and release of lactate deshydrogenase (LDH). Pro-inflammatory effects are estimated by ELISA in order to quantify IL-8 and MCP-1 secretions. Pro-oxidative effects are measured with hydroethydine (HE).

Results: Iron Texas Red nanoparticles accumulate at the cytoplasmic membrane level. They induce a slight LDH release, and have no inflammatory or oxidative effects. However, they enhance the cytotoxic, pro-inflammatory and oxidative effects of 7KC. The accumulation dynamics of SYTOX Green in cells is measured by CLSM to characterize the toxicity of nanoparticles. The emission spectra of SYTOX Green and nanoparticles are differentiated, and corresponding factor images specify the possible capture and cellular localization of nanoparticles in cells.

Conclusion: The designed protocol makes it possible to show how Iron Texas Red nanoparticles are captured by cardiomyocytes. Interestingly, whereas these fluorescent iron nanoparticles have no cytotoxic, pro-inflammatory or oxidative activities, they enhance the side effects of 7KC.

Show MeSH
Related in: MedlinePlus