Amelogenin splice isoforms stimulate chondrogenic differentiation of ATDC5 cells

K. Mitani, N. Haruyama, J. Hatakeyama, K. Igarashi

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Objective: Amelogenins are the most abundant matrix proteins in enamel. Among the amelogenin isoforms, full-length amelogenin (M180) and leucine-rich amelogenin peptide (LRAP) are expressed in various tissues and are implicated as signalling molecules in mesenchymal cells. Here, we examined the effects of M180 and LRAP on a chondrogenic cell line, ATDC5, to investigate the role of amelogenins in chondrogenesis. Materials and Methods: Recombinant mouse M180- or LRAP-protein-containing medium or control medium was mixed with a chondrogenesis-stimulating medium, and changes in the phenotype, gene expression levels and cell proliferation of cultured ATDC5 cells were analysed. Results: The addition of amelogenins increased alkaline phosphatase activity and glycosaminoglycan secretion at 14 and 21 days of culture, respectively, as compared with the control. Quantitative PCR (Q-PCR) analysis revealed that LRAP increased the gene expression levels of Runx2, Col2a1 and Aggrecan at 7 days of differentiation. Moreover, both M180 and LRAP significantly increased the gene expression levels of ALP, Aggrecan, Col10a1 and osteopontin at 28 days of culture. Bromodeoxyuridine assay and Q-PCR analysis for Wnt signalling indicated that both M180 and LRAP reduced proliferation, but induced the cell differentiation possibly through altered non-canonical Wnt signalling. Conclusion: M180 and LRAP accelerate chondrogenic differentiation and maturation of ATDC5 cells.

Original languageEnglish
Pages (from-to)169-179
Number of pages11
JournalOral Diseases
Volume19
Issue number2
DOIs
Publication statusPublished - Mar 1 2013
Externally publishedYes

Fingerprint

Amelogenin
Cell Differentiation
Protein Isoforms
Aggrecans
Chondrogenesis
Gene Expression
Polymerase Chain Reaction
Osteopontin
Bromodeoxyuridine
leucine-rich amelogenin peptide
Glycosaminoglycans
Alkaline Phosphatase
Cultured Cells
Cell Proliferation
Phenotype
Cell Line

All Science Journal Classification (ASJC) codes

  • Otorhinolaryngology
  • Dentistry(all)

Cite this

Amelogenin splice isoforms stimulate chondrogenic differentiation of ATDC5 cells. / Mitani, K.; Haruyama, N.; Hatakeyama, J.; Igarashi, K.

In: Oral Diseases, Vol. 19, No. 2, 01.03.2013, p. 169-179.

Research output: Contribution to journalArticle

Mitani, K. ; Haruyama, N. ; Hatakeyama, J. ; Igarashi, K. / Amelogenin splice isoforms stimulate chondrogenic differentiation of ATDC5 cells. In: Oral Diseases. 2013 ; Vol. 19, No. 2. pp. 169-179.
@article{5ce82886ab324ff4b198f0623eba96e9,
title = "Amelogenin splice isoforms stimulate chondrogenic differentiation of ATDC5 cells",
abstract = "Objective: Amelogenins are the most abundant matrix proteins in enamel. Among the amelogenin isoforms, full-length amelogenin (M180) and leucine-rich amelogenin peptide (LRAP) are expressed in various tissues and are implicated as signalling molecules in mesenchymal cells. Here, we examined the effects of M180 and LRAP on a chondrogenic cell line, ATDC5, to investigate the role of amelogenins in chondrogenesis. Materials and Methods: Recombinant mouse M180- or LRAP-protein-containing medium or control medium was mixed with a chondrogenesis-stimulating medium, and changes in the phenotype, gene expression levels and cell proliferation of cultured ATDC5 cells were analysed. Results: The addition of amelogenins increased alkaline phosphatase activity and glycosaminoglycan secretion at 14 and 21 days of culture, respectively, as compared with the control. Quantitative PCR (Q-PCR) analysis revealed that LRAP increased the gene expression levels of Runx2, Col2a1 and Aggrecan at 7 days of differentiation. Moreover, both M180 and LRAP significantly increased the gene expression levels of ALP, Aggrecan, Col10a1 and osteopontin at 28 days of culture. Bromodeoxyuridine assay and Q-PCR analysis for Wnt signalling indicated that both M180 and LRAP reduced proliferation, but induced the cell differentiation possibly through altered non-canonical Wnt signalling. Conclusion: M180 and LRAP accelerate chondrogenic differentiation and maturation of ATDC5 cells.",
author = "K. Mitani and N. Haruyama and J. Hatakeyama and K. Igarashi",
year = "2013",
month = "3",
day = "1",
doi = "10.1111/j.1601-0825.2012.01967.x",
language = "English",
volume = "19",
pages = "169--179",
journal = "Oral Diseases",
issn = "1354-523X",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Amelogenin splice isoforms stimulate chondrogenic differentiation of ATDC5 cells

AU - Mitani, K.

AU - Haruyama, N.

AU - Hatakeyama, J.

AU - Igarashi, K.

PY - 2013/3/1

Y1 - 2013/3/1

N2 - Objective: Amelogenins are the most abundant matrix proteins in enamel. Among the amelogenin isoforms, full-length amelogenin (M180) and leucine-rich amelogenin peptide (LRAP) are expressed in various tissues and are implicated as signalling molecules in mesenchymal cells. Here, we examined the effects of M180 and LRAP on a chondrogenic cell line, ATDC5, to investigate the role of amelogenins in chondrogenesis. Materials and Methods: Recombinant mouse M180- or LRAP-protein-containing medium or control medium was mixed with a chondrogenesis-stimulating medium, and changes in the phenotype, gene expression levels and cell proliferation of cultured ATDC5 cells were analysed. Results: The addition of amelogenins increased alkaline phosphatase activity and glycosaminoglycan secretion at 14 and 21 days of culture, respectively, as compared with the control. Quantitative PCR (Q-PCR) analysis revealed that LRAP increased the gene expression levels of Runx2, Col2a1 and Aggrecan at 7 days of differentiation. Moreover, both M180 and LRAP significantly increased the gene expression levels of ALP, Aggrecan, Col10a1 and osteopontin at 28 days of culture. Bromodeoxyuridine assay and Q-PCR analysis for Wnt signalling indicated that both M180 and LRAP reduced proliferation, but induced the cell differentiation possibly through altered non-canonical Wnt signalling. Conclusion: M180 and LRAP accelerate chondrogenic differentiation and maturation of ATDC5 cells.

AB - Objective: Amelogenins are the most abundant matrix proteins in enamel. Among the amelogenin isoforms, full-length amelogenin (M180) and leucine-rich amelogenin peptide (LRAP) are expressed in various tissues and are implicated as signalling molecules in mesenchymal cells. Here, we examined the effects of M180 and LRAP on a chondrogenic cell line, ATDC5, to investigate the role of amelogenins in chondrogenesis. Materials and Methods: Recombinant mouse M180- or LRAP-protein-containing medium or control medium was mixed with a chondrogenesis-stimulating medium, and changes in the phenotype, gene expression levels and cell proliferation of cultured ATDC5 cells were analysed. Results: The addition of amelogenins increased alkaline phosphatase activity and glycosaminoglycan secretion at 14 and 21 days of culture, respectively, as compared with the control. Quantitative PCR (Q-PCR) analysis revealed that LRAP increased the gene expression levels of Runx2, Col2a1 and Aggrecan at 7 days of differentiation. Moreover, both M180 and LRAP significantly increased the gene expression levels of ALP, Aggrecan, Col10a1 and osteopontin at 28 days of culture. Bromodeoxyuridine assay and Q-PCR analysis for Wnt signalling indicated that both M180 and LRAP reduced proliferation, but induced the cell differentiation possibly through altered non-canonical Wnt signalling. Conclusion: M180 and LRAP accelerate chondrogenic differentiation and maturation of ATDC5 cells.

UR - http://www.scopus.com/inward/record.url?scp=84878139343&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84878139343&partnerID=8YFLogxK

U2 - 10.1111/j.1601-0825.2012.01967.x

DO - 10.1111/j.1601-0825.2012.01967.x

M3 - Article

C2 - 22863294

AN - SCOPUS:84878139343

VL - 19

SP - 169

EP - 179

JO - Oral Diseases

JF - Oral Diseases

SN - 1354-523X

IS - 2

ER -