Cargando…
Cellular Zn depletion by metal ion chelators (TPEN, DTPA and chelex resin) and its application to osteoblastic MC3T3-E1 cells
Trace mineral studies involving metal ion chelators have been conducted in investigating the response of gene and protein expressions of certain cell lines but a few had really focused on how these metal ion chelators could affect the availability of important trace minerals such as Zn, Mn, Fe and C...
Autores principales: | , , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
The Korean Nutrition Society and The Korean Society of Community Nutrition
2007
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2882573/ https://www.ncbi.nlm.nih.gov/pubmed/20535382 http://dx.doi.org/10.4162/nrp.2007.1.1.29 |
_version_ | 1782182196702871552 |
---|---|
author | Cho, Young-Eun Lomeda, Ria-Ann R. Ryu, Sang-Hoon Lee, Jong-Hwa Beattie, John H. Kwun, In-Sook |
author_facet | Cho, Young-Eun Lomeda, Ria-Ann R. Ryu, Sang-Hoon Lee, Jong-Hwa Beattie, John H. Kwun, In-Sook |
author_sort | Cho, Young-Eun |
collection | PubMed |
description | Trace mineral studies involving metal ion chelators have been conducted in investigating the response of gene and protein expressions of certain cell lines but a few had really focused on how these metal ion chelators could affect the availability of important trace minerals such as Zn, Mn, Fe and Cu. The aim of the present study was to investigate the availability of Zn for the treatment of MC3T3-E1 osteoblast-like cells and the availability of some trace minerals in the cell culture media components after using chelexing resin in the FBS and the addition of N,N,N',N'-tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN, membrane-permeable chelator) and diethylenetriaminepentaacetic acid (DTPA, membrane-impermeable chelator) in the treatment medium. Components for the preparation of cell culture medium and Zn-treated medium have been tested for Zn, Mn, Fe and Cu contents by atomic absorption spectrophotometer or inductively coupled plasma spectrophotometer. Also, the expression of bone-related genes (ALP, Runx2, PTH-R, ProCOL I, OPN and OC) was measured on the cellular Zn depletion such as chelexing or TPEN treatment. Results have shown that using the chelexing resin in FBS would significantly decrease the available Zn (p<0.05) (39.4 ± 1.5 µM vs 0.61 ± 10.15 µM) and Mn (p<0.05) (0.74 ± 0.01 µM vs 0.12 ± 0.04 µM). However, levels of Fe and Cu in FBS were not changed by chelexing FBS. The use of TPEN and DTPA as Zn-chelators did not show significant difference on the final concentration of Zn in the treatment medium (0, 3, 6, 9, 12 µM) except for in the addition of higher 15 µM ZnCl(2) which showed a significant increase of Zn level in DTPA-chelated treatment medium. Results have shown that both chelators gave the same pattern for the expression of the five bone-related genes between Zn- and Zn+, and TPEN-treated experiments, compared to chelex-treated experiment, showed lower bone-related gene expression, which may imply that TPEN would be a stronger chelator than chelex resin. This study showed that TPEN would be a stronger chelator compared to DTPA or chelex resin and TPEN and chelex resin exerted cellular zinc depletion to be enough for cell study for Zn depletion. |
format | Text |
id | pubmed-2882573 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | The Korean Nutrition Society and The Korean Society of Community Nutrition |
record_format | MEDLINE/PubMed |
spelling | pubmed-28825732010-06-09 Cellular Zn depletion by metal ion chelators (TPEN, DTPA and chelex resin) and its application to osteoblastic MC3T3-E1 cells Cho, Young-Eun Lomeda, Ria-Ann R. Ryu, Sang-Hoon Lee, Jong-Hwa Beattie, John H. Kwun, In-Sook Nutr Res Pract Original Research Trace mineral studies involving metal ion chelators have been conducted in investigating the response of gene and protein expressions of certain cell lines but a few had really focused on how these metal ion chelators could affect the availability of important trace minerals such as Zn, Mn, Fe and Cu. The aim of the present study was to investigate the availability of Zn for the treatment of MC3T3-E1 osteoblast-like cells and the availability of some trace minerals in the cell culture media components after using chelexing resin in the FBS and the addition of N,N,N',N'-tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN, membrane-permeable chelator) and diethylenetriaminepentaacetic acid (DTPA, membrane-impermeable chelator) in the treatment medium. Components for the preparation of cell culture medium and Zn-treated medium have been tested for Zn, Mn, Fe and Cu contents by atomic absorption spectrophotometer or inductively coupled plasma spectrophotometer. Also, the expression of bone-related genes (ALP, Runx2, PTH-R, ProCOL I, OPN and OC) was measured on the cellular Zn depletion such as chelexing or TPEN treatment. Results have shown that using the chelexing resin in FBS would significantly decrease the available Zn (p<0.05) (39.4 ± 1.5 µM vs 0.61 ± 10.15 µM) and Mn (p<0.05) (0.74 ± 0.01 µM vs 0.12 ± 0.04 µM). However, levels of Fe and Cu in FBS were not changed by chelexing FBS. The use of TPEN and DTPA as Zn-chelators did not show significant difference on the final concentration of Zn in the treatment medium (0, 3, 6, 9, 12 µM) except for in the addition of higher 15 µM ZnCl(2) which showed a significant increase of Zn level in DTPA-chelated treatment medium. Results have shown that both chelators gave the same pattern for the expression of the five bone-related genes between Zn- and Zn+, and TPEN-treated experiments, compared to chelex-treated experiment, showed lower bone-related gene expression, which may imply that TPEN would be a stronger chelator than chelex resin. This study showed that TPEN would be a stronger chelator compared to DTPA or chelex resin and TPEN and chelex resin exerted cellular zinc depletion to be enough for cell study for Zn depletion. The Korean Nutrition Society and The Korean Society of Community Nutrition 2007 2007-03-31 /pmc/articles/PMC2882573/ /pubmed/20535382 http://dx.doi.org/10.4162/nrp.2007.1.1.29 Text en ©2007 The Korean Nutrition Society and The Korean Society of Community Nutrition http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Research Cho, Young-Eun Lomeda, Ria-Ann R. Ryu, Sang-Hoon Lee, Jong-Hwa Beattie, John H. Kwun, In-Sook Cellular Zn depletion by metal ion chelators (TPEN, DTPA and chelex resin) and its application to osteoblastic MC3T3-E1 cells |
title | Cellular Zn depletion by metal ion chelators (TPEN, DTPA and chelex resin) and its application to osteoblastic MC3T3-E1 cells |
title_full | Cellular Zn depletion by metal ion chelators (TPEN, DTPA and chelex resin) and its application to osteoblastic MC3T3-E1 cells |
title_fullStr | Cellular Zn depletion by metal ion chelators (TPEN, DTPA and chelex resin) and its application to osteoblastic MC3T3-E1 cells |
title_full_unstemmed | Cellular Zn depletion by metal ion chelators (TPEN, DTPA and chelex resin) and its application to osteoblastic MC3T3-E1 cells |
title_short | Cellular Zn depletion by metal ion chelators (TPEN, DTPA and chelex resin) and its application to osteoblastic MC3T3-E1 cells |
title_sort | cellular zn depletion by metal ion chelators (tpen, dtpa and chelex resin) and its application to osteoblastic mc3t3-e1 cells |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2882573/ https://www.ncbi.nlm.nih.gov/pubmed/20535382 http://dx.doi.org/10.4162/nrp.2007.1.1.29 |
work_keys_str_mv | AT choyoungeun cellularzndepletionbymetalionchelatorstpendtpaandchelexresinanditsapplicationtoosteoblasticmc3t3e1cells AT lomedariaannr cellularzndepletionbymetalionchelatorstpendtpaandchelexresinanditsapplicationtoosteoblasticmc3t3e1cells AT ryusanghoon cellularzndepletionbymetalionchelatorstpendtpaandchelexresinanditsapplicationtoosteoblasticmc3t3e1cells AT leejonghwa cellularzndepletionbymetalionchelatorstpendtpaandchelexresinanditsapplicationtoosteoblasticmc3t3e1cells AT beattiejohnh cellularzndepletionbymetalionchelatorstpendtpaandchelexresinanditsapplicationtoosteoblasticmc3t3e1cells AT kwuninsook cellularzndepletionbymetalionchelatorstpendtpaandchelexresinanditsapplicationtoosteoblasticmc3t3e1cells |