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Silver wire amplifies the signaling mechanism for IL-1beta production more than silver submicroparticles in human monocytic THP-1 cells.

Jung HJ, Pak PJ, Park SH, Ju JE, Kim JS, Lee HS, Chung N - PLoS ONE (2014)

Bottom Line: Silver materials have been widely used in diverse fields.This trend was also observed with each step of the signaling mechanism for IL-1β production, which is a single pathway affiliated with ROS generation or lysosomal rupture or both, cathepsin B, caspase-1 (NALP3 inflammasome), and finally IL-1β production in THP-1 cells.All these results suggest that, for development of safe and effective silver materials, the shape or form of silver materials should be considered, especially for macrophage cell lines because epithelial cell lines are not overly sensitive to silver materials.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosystems and Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, Korea.

ABSTRACT
Silver materials have been widely used in diverse fields. However, their toxicity and their mechanism, especially in different forms, have not been studied sufficiently. Thus, cytotoxicity, apoptosis, and interleukin-1beta (IL-1β) production were investigated using macrophage-like THP-1 cells in the presence of Ag microparticles (AgMPs, 2.7 µm), Ag submicroparticles (AgSMPs, 150 nm), and Ag wires (AgWs, 274 nm×5.3 µm). The levels of cytotoxicity, apoptosis, and IL-1β production by AgWs were higher than those by the other two AgSMPs and AgMPs. This trend was also observed with each step of the signaling mechanism for IL-1β production, which is a single pathway affiliated with ROS generation or lysosomal rupture or both, cathepsin B, caspase-1 (NALP3 inflammasome), and finally IL-1β production in THP-1 cells. All these results suggest that, for development of safe and effective silver materials, the shape or form of silver materials should be considered, especially for macrophage cell lines because epithelial cell lines are not overly sensitive to silver materials.

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Involvement of lysosomal destabilization and cathepsin B activity with IL-1β production in the presence of silver materials.(A) PMA-primed THP-1 cells were treated with each of the silver materials (100 µg/mL) for 24 h. At the end of exposure, the cells were loaded with acridine orange (20 µg/mL) for 15 min at 37°C and photographed using a fluorescence microscope (400×). (B) Lysosomal destabilization as measured by loss of fluorescence with increasing concentration of silver materials. The cells were preloaded with acridine orange (0.5 µg/mL) for 30 min, treated with silver materials, and analyzed using FACS to determine the loss of acridine orange from lysosome. Data are represented as the mean ±SD (n = 3). (C) Involvement of cathepsin B in silver materials-induced IL-1β production. The cells were treated with different forms of silver materials for 24 h in both the absence and presence of cathepsin B inhibitor (CA-074-Me; 10 µM). IL-1β levels were measured using ELISA. Data are represented as the mean ±SD (n = 3).
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pone-0112256-g006: Involvement of lysosomal destabilization and cathepsin B activity with IL-1β production in the presence of silver materials.(A) PMA-primed THP-1 cells were treated with each of the silver materials (100 µg/mL) for 24 h. At the end of exposure, the cells were loaded with acridine orange (20 µg/mL) for 15 min at 37°C and photographed using a fluorescence microscope (400×). (B) Lysosomal destabilization as measured by loss of fluorescence with increasing concentration of silver materials. The cells were preloaded with acridine orange (0.5 µg/mL) for 30 min, treated with silver materials, and analyzed using FACS to determine the loss of acridine orange from lysosome. Data are represented as the mean ±SD (n = 3). (C) Involvement of cathepsin B in silver materials-induced IL-1β production. The cells were treated with different forms of silver materials for 24 h in both the absence and presence of cathepsin B inhibitor (CA-074-Me; 10 µM). IL-1β levels were measured using ELISA. Data are represented as the mean ±SD (n = 3).

Mentions: The activator of NALP3 inflammasome, for example crystalline or particulate materials, causes phagocytosis and then leads to lysosomal damage, resulting in cytosolic release of lysosomal contents [8]. Cathepsin B, a lysosomal protease, triggers the NALP3 inflammasome directly. To evaluate the lysosomal destabilization by silver materials, cell staining was conducted with acridine orange and pictured using a fluorescence microscope (Fig. 6A). Acridine orange reacts with the acidic content of organelles after the silver materials incurs membrane damage and then the fluorescence intensity decreases. As can be observed in Fig. 6A, the fluorescent intensity with AgWs was lower or weaker than that with the others. The fluorescent intensity of PMA-primed THP-1 cells treated with silver materials showed a very similar trend (Fig. 6B). These results demonstrate that different forms of silver materials induced the destabilization of lysosomal membranes dose-dependently and that AgWs caused more lysosomal membrane damage than the others.


Silver wire amplifies the signaling mechanism for IL-1beta production more than silver submicroparticles in human monocytic THP-1 cells.

Jung HJ, Pak PJ, Park SH, Ju JE, Kim JS, Lee HS, Chung N - PLoS ONE (2014)

Involvement of lysosomal destabilization and cathepsin B activity with IL-1β production in the presence of silver materials.(A) PMA-primed THP-1 cells were treated with each of the silver materials (100 µg/mL) for 24 h. At the end of exposure, the cells were loaded with acridine orange (20 µg/mL) for 15 min at 37°C and photographed using a fluorescence microscope (400×). (B) Lysosomal destabilization as measured by loss of fluorescence with increasing concentration of silver materials. The cells were preloaded with acridine orange (0.5 µg/mL) for 30 min, treated with silver materials, and analyzed using FACS to determine the loss of acridine orange from lysosome. Data are represented as the mean ±SD (n = 3). (C) Involvement of cathepsin B in silver materials-induced IL-1β production. The cells were treated with different forms of silver materials for 24 h in both the absence and presence of cathepsin B inhibitor (CA-074-Me; 10 µM). IL-1β levels were measured using ELISA. Data are represented as the mean ±SD (n = 3).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4232372&req=5

pone-0112256-g006: Involvement of lysosomal destabilization and cathepsin B activity with IL-1β production in the presence of silver materials.(A) PMA-primed THP-1 cells were treated with each of the silver materials (100 µg/mL) for 24 h. At the end of exposure, the cells were loaded with acridine orange (20 µg/mL) for 15 min at 37°C and photographed using a fluorescence microscope (400×). (B) Lysosomal destabilization as measured by loss of fluorescence with increasing concentration of silver materials. The cells were preloaded with acridine orange (0.5 µg/mL) for 30 min, treated with silver materials, and analyzed using FACS to determine the loss of acridine orange from lysosome. Data are represented as the mean ±SD (n = 3). (C) Involvement of cathepsin B in silver materials-induced IL-1β production. The cells were treated with different forms of silver materials for 24 h in both the absence and presence of cathepsin B inhibitor (CA-074-Me; 10 µM). IL-1β levels were measured using ELISA. Data are represented as the mean ±SD (n = 3).
Mentions: The activator of NALP3 inflammasome, for example crystalline or particulate materials, causes phagocytosis and then leads to lysosomal damage, resulting in cytosolic release of lysosomal contents [8]. Cathepsin B, a lysosomal protease, triggers the NALP3 inflammasome directly. To evaluate the lysosomal destabilization by silver materials, cell staining was conducted with acridine orange and pictured using a fluorescence microscope (Fig. 6A). Acridine orange reacts with the acidic content of organelles after the silver materials incurs membrane damage and then the fluorescence intensity decreases. As can be observed in Fig. 6A, the fluorescent intensity with AgWs was lower or weaker than that with the others. The fluorescent intensity of PMA-primed THP-1 cells treated with silver materials showed a very similar trend (Fig. 6B). These results demonstrate that different forms of silver materials induced the destabilization of lysosomal membranes dose-dependently and that AgWs caused more lysosomal membrane damage than the others.

Bottom Line: Silver materials have been widely used in diverse fields.This trend was also observed with each step of the signaling mechanism for IL-1β production, which is a single pathway affiliated with ROS generation or lysosomal rupture or both, cathepsin B, caspase-1 (NALP3 inflammasome), and finally IL-1β production in THP-1 cells.All these results suggest that, for development of safe and effective silver materials, the shape or form of silver materials should be considered, especially for macrophage cell lines because epithelial cell lines are not overly sensitive to silver materials.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosystems and Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, Korea.

ABSTRACT
Silver materials have been widely used in diverse fields. However, their toxicity and their mechanism, especially in different forms, have not been studied sufficiently. Thus, cytotoxicity, apoptosis, and interleukin-1beta (IL-1β) production were investigated using macrophage-like THP-1 cells in the presence of Ag microparticles (AgMPs, 2.7 µm), Ag submicroparticles (AgSMPs, 150 nm), and Ag wires (AgWs, 274 nm×5.3 µm). The levels of cytotoxicity, apoptosis, and IL-1β production by AgWs were higher than those by the other two AgSMPs and AgMPs. This trend was also observed with each step of the signaling mechanism for IL-1β production, which is a single pathway affiliated with ROS generation or lysosomal rupture or both, cathepsin B, caspase-1 (NALP3 inflammasome), and finally IL-1β production in THP-1 cells. All these results suggest that, for development of safe and effective silver materials, the shape or form of silver materials should be considered, especially for macrophage cell lines because epithelial cell lines are not overly sensitive to silver materials.

Show MeSH
Related in: MedlinePlus