Skip to main content
SpringerLink
Log in

Role of metallothionein isoforms in bone formation processes in rat marrow mesenchymal stem cells in culture

  • Original Articles
  • Published:
Biological Trace Element Research Aims and scope Submit manuscript

Abstract

Temporal changes in mRNAs for metallothionein (MT) isoforms in subcultures of rat marrow mesenchymal stem cells (MSCs) after treatment with dexamethasone were investigated. Both MT-1 and MT-2 mRNA expression in the cultured MSCs with dexamethasone showed maximum levels at d 1, whereas ALP and osteocalcin mRNAs peaked at d 12. MT-3 mRNA was not detected in the cultured MSCs at any time. The expression level of MT-2 mRNA at d 1 was 9.4-fold higher than that of MT-1 mRNA. Finally, osteoblast differentiation and mineralization of MSCs at d 14 was inhibited by the addition of a common antisense oligonucleotide for both MT-1 and MT-2 in the culture medium during the first 4 d. The results suggest that the large amounts of MT-2 are produced in the early stage of subculture of MSCs, and this might regulate their differentiation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. D. L. Diefenderfer, A. M. Osyczka, G. C. Reilly, and P. S. Leboy, BMP responsiveness in human mesenchymal stem cells, Connect. Tissue Res. 44, 305–311 (2003).

    Article  PubMed  CAS  Google Scholar 

  2. R. Sordella, W. Jiang, G.-C. Chen, M. Curto, and J. Settleman, Modulation of Rho GTPase signaling regulates a switch between adipogenesis and myogenesis, Cell, 113, 147–158 (2003).

    Article  PubMed  CAS  Google Scholar 

  3. N. watanabe, Y. Tezuka, K. Matsuno, et al., Suppression of differentiation and proliferation of early chondrogenic cells by Notch, J. Bone Miner. Metab., 21, 344–352 (2003).

    Article  PubMed  CAS  Google Scholar 

  4. H. Ohgushi, Y. Dohi, S. Tamai, and S. Tabata, Osteogenic differentiation of marrow stromal stem cells in porous hydroxyapatite ceramics, J. Biomed. Mater. Res. 27, 1401–1407 (1993).

    Article  PubMed  CAS  Google Scholar 

  5. H. Ohgushi, Y. Dohi, T. Katuda, S. Tamai, S. Tabata, and Y. Suwa, In vitro bone formation by rat marrow cell culture, J. Biomed. Mater. Res., 32, 333–340 (1996).

    Article  PubMed  CAS  Google Scholar 

  6. H. Ohgushi and A. I. Caplan, Stem cell technology and bioceramics: from cell to gene engineering, J. Biomed. Mater. Res. 48, 913–927 (1999).

    Article  PubMed  CAS  Google Scholar 

  7. T. Yoshikawa, H. Ohgushi, and S. Tamai, Immediate bone forming capability of prefabricated osteogenic hydroxyapatite, J. Biomed. Mater. Res. 32, 481–492 (1996).

    Article  PubMed  CAS  Google Scholar 

  8. H. Ohgushi, T. Yoshikawa, H. Nakajima, S. Tamai, Y. Dohi, and K. Okunaga, Al2O3 doped apatite-wollastonite containing glass ceramic provokes osteogenic differentiation of marrow stromal stem cells, J. Biomed. Mater. Res. 44, 381–388 (1999).

    Article  PubMed  CAS  Google Scholar 

  9. P. Moffatt and F. Denizeau, Metallothionein in physiological and physiopathological processes, Drug Metab. Rev., 29, 261–307 (1997).

    PubMed  CAS  Google Scholar 

  10. A. T. Miles, G. M. Hawksworth, J. H. Beattie, and V. Rodilla, Induction, regulation, degradation, and biological significance of mammalian metallothioneins, Crit. Rev. Biochem. Mol. Biol. 35, 35–70 (2000).

    Article  PubMed  CAS  Google Scholar 

  11. T. Kawashima, K. Doh-ura, M. Torisu, Y. Uchida, A. Furuta, and T. Iwaki, Differential expression of metallothioneins in human prion diseases, Dement. Geriatr. Gogn. Disord. 11, 251–262 (2000).

    Article  CAS  Google Scholar 

  12. T. Minami, S. Ichida, and K. Kubo, Study of metallothionein using capaillary zone electrophoresis, J. Chromatogr. B 781, 303–311 (2002).

    Article  CAS  Google Scholar 

  13. Y. Dohi, K. Sugimoto, T. Yoshikawa, et al., Effect of cadmium on osteogenesis within diffusion chambers by bone marrow cells: biochemical evidence of decreased bone formation capacity, Toxicol. Appl. Pharmacol. 120, 274–280 (1993).

    Article  PubMed  CAS  Google Scholar 

  14. T. Miyahara, M. Nemoto, S.-I. Tukamoto, et al., Induction of metallothionen and stimulation of calcification by dexamethasone in cultured clonal osteogenic cells, MC3T3-E1, Toxicol. Lett. 57, 257–267 (1991).

    Article  PubMed  CAS  Google Scholar 

  15. C. Maniatopoulos, J. Sodek, and A. H. Melcher, Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats, Cell Tissue Res. 254, 317–330 (1988).

    Article  PubMed  CAS  Google Scholar 

  16. H. Shimaoka, Y. Dohi, H. Ohgushi, et al., Recombinant growth/differentiation factor-5 (GDF-5) stimulates osteogenic differentiation of marrow mesenchymal stem cells in porous hydroxyapatite ceramic, J. Biomed. Mater. Res. 68A, 168–176 (2004).

    Article  CAS  Google Scholar 

  17. Y. Dohi, H. Nakajima, H. Ohgushi, and K. Yonemasu, Quantification of matrix protein mRNAs expression during mineralized tissue formation in rat marrow cell culture by a real time quantitative PCR method, Key Eng. Mater. 192–195, 367–372 (2001).

    Article  Google Scholar 

  18. Y. Dohi, H. Ohgushi, T. Minami, M. Ikeuchi, and K. Yonemasu, Quantification of rat and mouse metallothionein-I mRNA expression by a real-time quantitative PCR using a common fluorogenic probe, Recent Res. Devel. Anal. Biochem. 2, 95–106 (2002).

    CAS  Google Scholar 

  19. T. Yoshikawa, H. Ohgushi, M. Akahane, S. Tamai, and K. Ichijima, Analysis of gene expression in osteogenic cultured marrow/hydroxyapatite construct implanted at ectopic sites: a comparison with the osteogenic ability of cancellous bone, J. Biomed. Mater. Res. 41, 568–573 (1998).

    Article  PubMed  CAS  Google Scholar 

  20. T. Yoshikawa, H. Ohgushi, H. Nakajima, et al., In vivo osteogenetic durability of cultured bone in porous ceramics: a novel method for autogenous bone graft substitution, Transplantation 69, 128–134 (2000).

    Article  PubMed  CAS  Google Scholar 

  21. J. M. DeMoor, W. A. Kennette, O. M. Collins, and J. Koropatnick, Zinc-metallothionein levels are correlated with enhanced glucocorticoid responsiveness in mouse cells exposed to ZnC12, HgC12, and heat shock, Toxicol. Sci. 64, 67–76 (2001).

    PubMed  CAS  Google Scholar 

  22. M. Yamaguchi and T. Matsu, Stimulatory effect of zinc-chelating dipeptide on deoxyribonucleic acid synthesis in osteoblastic MC3T3-E1 cells, Peptides 17, 1207–1211 (1996).

    Article  PubMed  CAS  Google Scholar 

  23. A. Igarashi and M. Yamaguchi, Great increase in bone 66 kDa protein and osteocalcin at later stages with healing rat fractures: effect of zinc treatment, Int. J. Mol. Med. 11, 223–228 (2003).

    PubMed  CAS  Google Scholar 

  24. H. Kawamura, A. Ito, T. Muramatsu, S. Miyakawa, N. Ochiai, and T. Tateishi Long-term implantation of zinc-releasing calcium phosphate ceramics in rabbit femora, J. Biomed. Mater. Res. 65A, 468–474 (2003).

    Article  CAS  Google Scholar 

  25. Y. Sogo, T. Sakurai, K. Onuma, and A. Ito, The most appropriate (Ca+Zn)/P molar ratio to minimize the zinc content of ZnTCP/HAP ceramic used in the promotion of bone formation, J. Biomed. Mater. Res. 62, 457–463 (2002).

    Article  PubMed  CAS  Google Scholar 

  26. H. Kawamura, A. Ito, S. Miyakawa, et al., Stimulatory effect of zinc-releasing calcium phosphate implant on bone formation in rabbit femora, J. Biomed. Mater. Res. 50, 184–190 (2000).

    Article  PubMed  CAS  Google Scholar 

  27. D. Beyersmann and H. Haase, Functions of zinc in signaling, proliferation and differentiation of mammalian cells, Biometals 14, 331–341 (2001).

    Article  PubMed  CAS  Google Scholar 

  28. J. Cao, J. A. Bobo, J. P. Liuzzi, and R. J. Cousins, Effects of intracellular zinc depletion on metallothionein and ZIP2 transporter expression and apoptosis, J. Leukocyte Biol. 70, 559–566 (2001).

    PubMed  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Takeshi Minami

    Present address: Department of Life Sciences, School of Science and Technology, Kinki University, 577-8502, Higashi-osaka, Japan

Authors and Affiliations

  1. Department of Public Health, Nara Medical University, 840 Shijo-cho, 634-8521, Kashihara City, Nara, Japan

    Yoshiko Dohi, Hideki Shimaoka & Kunio Yonemasu

  2. Department of Oral and Maxillofacial Surgery, Nara Medical University, 840 Shijo-cho, 634-8521, Kashihara City, Nara, Japan

    Masako Ikeuchi

  3. Department of Tissue Engineering Research Center, National Institute of Advanced Industrial Science and Technology, Amagasaki Site 3-11-46 Nakouji, 661-0974, Amagasaki City, Hyogo, Japan

    Hajime Ohgushi

Authors
  1. Yoshiko Dohi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hideki Shimaoka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Masako Ikeuchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hajime Ohgushi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kunio Yonemasu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Takeshi Minami
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dohi, Y., Shimaoka, H., Ikeuchi, M. et al. Role of metallothionein isoforms in bone formation processes in rat marrow mesenchymal stem cells in culture. Biol Trace Elem Res 104, 57–69 (2005). https://doi.org/10.1385/BTER:104:1:057

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1385/BTER:104:1:057

Index Entries

Search

Navigation