Osteoblast protects osteoclast devoid of sodium-dependent vitamin C transporters from oxidative cytotoxicity of ascorbic acid

Takeshi Takarada, Eiichi Hinoi, Yuki Kambe, Koichi Sahara, Shintaro Kurokawa, Yoshifumi Takahata, Yukio Yoneda

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The view that ascorbic acid indirectly benefits osteoclastogenesis through expression of receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL) by osteoblasts is prevailing. In this study, we have examined the direct effect of ascorbic acid on osteoclastogenesis in cultured mouse osteoclasts differentiated from bone marrow precursors. The absence of alkaline phosphatase and osteoblastic marker genes validated the usefulness of isolation procedures. Sustained exposure to ascorbic acid, but not to dehydroascorbic acid, significantly reduced the number of multinucleated cells positive to tartrate-resistant acid phosphatase (TRAP) staining. In cultured osteoclasts, mRNA expression was seen for glucose transporter-1 involved in membrane transport of dehydroascorbic acid, but not for sodium-dependent vitamin C transporters-1 and -2 that are both responsible for the transport of ascorbic acid. The inhibition by ascorbic acid was completely prevented by catalase, while ascorbic acid or hydrogen peroxide drastically increased the number of cells stained with propidium iodide and the generation of reactive oxygen species, in addition to inducing mitochondrial membrane depolarization in cultured osteoclasts. In pre-osteoclastic cell line RAW264.7 cells, ascorbic acid similarly inhibited the formation of TRAP-positive multinucleated cells, with a significant decrease in RANKL-induced NF-κB transactivation. Moreover, co-culture with osteoblastic MC3T3-E1 cells significantly prevented the ascorbic acid-induced decrease in the number of TRAP-positive multinucleated cells in RAW264.7 cells. These results suggest that ascorbic acid may play a dual repulsive role in osteoclastogenesis toward bone remodeling through the direct cytotoxicity mediated by oxidative stress to osteoclasts, in addition to the indirect trophism mediated by RANKL from osteoblasts.

Original languageEnglish
Pages (from-to)1-11
Number of pages11
JournalEuropean Journal of Pharmacology
Volume575
Issue number1-3
DOIs
Publication statusPublished - Dec 1 2007
Externally publishedYes

Fingerprint

Sodium-Coupled Vitamin C Transporters
Osteoclasts
Osteoblasts
Ascorbic Acid
Dehydroascorbic Acid
Osteogenesis
Cell Count
Facilitative Glucose Transport Proteins
Propidium
Bone Remodeling
Mitochondrial Membranes
Cytoplasmic and Nuclear Receptors
Coculture Techniques
Catalase
Hydrogen Peroxide
Transcriptional Activation
Alkaline Phosphatase
Reactive Oxygen Species

Keywords

  • Ascorbic acid
  • Catalase
  • Hydrogen peroxide
  • NF-κB
  • Osteoclasts
  • RANKL

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Pharmacology

Cite this

Osteoblast protects osteoclast devoid of sodium-dependent vitamin C transporters from oxidative cytotoxicity of ascorbic acid. / Takarada, Takeshi; Hinoi, Eiichi; Kambe, Yuki; Sahara, Koichi; Kurokawa, Shintaro; Takahata, Yoshifumi; Yoneda, Yukio.

In: European Journal of Pharmacology, Vol. 575, No. 1-3, 01.12.2007, p. 1-11.

Research output: Contribution to journalArticle

Takarada, Takeshi ; Hinoi, Eiichi ; Kambe, Yuki ; Sahara, Koichi ; Kurokawa, Shintaro ; Takahata, Yoshifumi ; Yoneda, Yukio. / Osteoblast protects osteoclast devoid of sodium-dependent vitamin C transporters from oxidative cytotoxicity of ascorbic acid. In: European Journal of Pharmacology. 2007 ; Vol. 575, No. 1-3. pp. 1-11.
@article{84eb9a8e02984df9b606bea6fb4b795f,
title = "Osteoblast protects osteoclast devoid of sodium-dependent vitamin C transporters from oxidative cytotoxicity of ascorbic acid",
abstract = "The view that ascorbic acid indirectly benefits osteoclastogenesis through expression of receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL) by osteoblasts is prevailing. In this study, we have examined the direct effect of ascorbic acid on osteoclastogenesis in cultured mouse osteoclasts differentiated from bone marrow precursors. The absence of alkaline phosphatase and osteoblastic marker genes validated the usefulness of isolation procedures. Sustained exposure to ascorbic acid, but not to dehydroascorbic acid, significantly reduced the number of multinucleated cells positive to tartrate-resistant acid phosphatase (TRAP) staining. In cultured osteoclasts, mRNA expression was seen for glucose transporter-1 involved in membrane transport of dehydroascorbic acid, but not for sodium-dependent vitamin C transporters-1 and -2 that are both responsible for the transport of ascorbic acid. The inhibition by ascorbic acid was completely prevented by catalase, while ascorbic acid or hydrogen peroxide drastically increased the number of cells stained with propidium iodide and the generation of reactive oxygen species, in addition to inducing mitochondrial membrane depolarization in cultured osteoclasts. In pre-osteoclastic cell line RAW264.7 cells, ascorbic acid similarly inhibited the formation of TRAP-positive multinucleated cells, with a significant decrease in RANKL-induced NF-κB transactivation. Moreover, co-culture with osteoblastic MC3T3-E1 cells significantly prevented the ascorbic acid-induced decrease in the number of TRAP-positive multinucleated cells in RAW264.7 cells. These results suggest that ascorbic acid may play a dual repulsive role in osteoclastogenesis toward bone remodeling through the direct cytotoxicity mediated by oxidative stress to osteoclasts, in addition to the indirect trophism mediated by RANKL from osteoblasts.",
keywords = "Ascorbic acid, Catalase, Hydrogen peroxide, NF-κB, Osteoclasts, RANKL",
author = "Takeshi Takarada and Eiichi Hinoi and Yuki Kambe and Koichi Sahara and Shintaro Kurokawa and Yoshifumi Takahata and Yukio Yoneda",
year = "2007",
month = "12",
day = "1",
doi = "10.1016/j.ejphar.2007.07.041",
language = "English",
volume = "575",
pages = "1--11",
journal = "European Journal of Pharmacology",
issn = "0014-2999",
publisher = "Elsevier",
number = "1-3",

}

TY - JOUR

T1 - Osteoblast protects osteoclast devoid of sodium-dependent vitamin C transporters from oxidative cytotoxicity of ascorbic acid

AU - Takarada, Takeshi

AU - Hinoi, Eiichi

AU - Kambe, Yuki

AU - Sahara, Koichi

AU - Kurokawa, Shintaro

AU - Takahata, Yoshifumi

AU - Yoneda, Yukio

PY - 2007/12/1

Y1 - 2007/12/1

N2 - The view that ascorbic acid indirectly benefits osteoclastogenesis through expression of receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL) by osteoblasts is prevailing. In this study, we have examined the direct effect of ascorbic acid on osteoclastogenesis in cultured mouse osteoclasts differentiated from bone marrow precursors. The absence of alkaline phosphatase and osteoblastic marker genes validated the usefulness of isolation procedures. Sustained exposure to ascorbic acid, but not to dehydroascorbic acid, significantly reduced the number of multinucleated cells positive to tartrate-resistant acid phosphatase (TRAP) staining. In cultured osteoclasts, mRNA expression was seen for glucose transporter-1 involved in membrane transport of dehydroascorbic acid, but not for sodium-dependent vitamin C transporters-1 and -2 that are both responsible for the transport of ascorbic acid. The inhibition by ascorbic acid was completely prevented by catalase, while ascorbic acid or hydrogen peroxide drastically increased the number of cells stained with propidium iodide and the generation of reactive oxygen species, in addition to inducing mitochondrial membrane depolarization in cultured osteoclasts. In pre-osteoclastic cell line RAW264.7 cells, ascorbic acid similarly inhibited the formation of TRAP-positive multinucleated cells, with a significant decrease in RANKL-induced NF-κB transactivation. Moreover, co-culture with osteoblastic MC3T3-E1 cells significantly prevented the ascorbic acid-induced decrease in the number of TRAP-positive multinucleated cells in RAW264.7 cells. These results suggest that ascorbic acid may play a dual repulsive role in osteoclastogenesis toward bone remodeling through the direct cytotoxicity mediated by oxidative stress to osteoclasts, in addition to the indirect trophism mediated by RANKL from osteoblasts.

AB - The view that ascorbic acid indirectly benefits osteoclastogenesis through expression of receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL) by osteoblasts is prevailing. In this study, we have examined the direct effect of ascorbic acid on osteoclastogenesis in cultured mouse osteoclasts differentiated from bone marrow precursors. The absence of alkaline phosphatase and osteoblastic marker genes validated the usefulness of isolation procedures. Sustained exposure to ascorbic acid, but not to dehydroascorbic acid, significantly reduced the number of multinucleated cells positive to tartrate-resistant acid phosphatase (TRAP) staining. In cultured osteoclasts, mRNA expression was seen for glucose transporter-1 involved in membrane transport of dehydroascorbic acid, but not for sodium-dependent vitamin C transporters-1 and -2 that are both responsible for the transport of ascorbic acid. The inhibition by ascorbic acid was completely prevented by catalase, while ascorbic acid or hydrogen peroxide drastically increased the number of cells stained with propidium iodide and the generation of reactive oxygen species, in addition to inducing mitochondrial membrane depolarization in cultured osteoclasts. In pre-osteoclastic cell line RAW264.7 cells, ascorbic acid similarly inhibited the formation of TRAP-positive multinucleated cells, with a significant decrease in RANKL-induced NF-κB transactivation. Moreover, co-culture with osteoblastic MC3T3-E1 cells significantly prevented the ascorbic acid-induced decrease in the number of TRAP-positive multinucleated cells in RAW264.7 cells. These results suggest that ascorbic acid may play a dual repulsive role in osteoclastogenesis toward bone remodeling through the direct cytotoxicity mediated by oxidative stress to osteoclasts, in addition to the indirect trophism mediated by RANKL from osteoblasts.

KW - Ascorbic acid

KW - Catalase

KW - Hydrogen peroxide

KW - NF-κB

KW - Osteoclasts

KW - RANKL

UR - http://www.scopus.com/inward/record.url?scp=35348853867&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=35348853867&partnerID=8YFLogxK

U2 - 10.1016/j.ejphar.2007.07.041

DO - 10.1016/j.ejphar.2007.07.041

M3 - Article

C2 - 17698058

AN - SCOPUS:35348853867

VL - 575

SP - 1

EP - 11

JO - European Journal of Pharmacology

JF - European Journal of Pharmacology

SN - 0014-2999

IS - 1-3

ER -