Oestrogen receptor β (ERβ) regulates osteogenic differentiation of human dental pulp cells

Aishah Alhodhodi, Hanaa Alkharobi, Matthew Humphries, Hasanain Alkhafaji, Reem El-Gendy, Georg Feichtinger, Valerie Speirs, James Beattie

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Estradiol (E2) has many important actions in the tissues of the oral cavity. Disruption of E2 metabolism or alterations in systemic E2 concentrations have been associated with compromised periodontal health. In many instances such changes occur secondarily to the well characterised effects of E2 on bone physiology -especially maintenance of bone mineral density (BMD). Despite these important epidemiological findings, little is known about the mechanism of action of E2 in oral tissues or the expression and function of oestrogen receptor (ER) isoforms in these tissues. We have isolated human dental pulp cells (hDPCs), which are able to differentiate towards an osteogenic lineage under appropriate culture conditions. We show that hDPCs express ERα, ERβ1, ERβ2 and the cell membrane associated G protein-coupled ER (GPR30). Following osteogenic differentiation of hDPCs, ERβ1 and ERβ2 were up regulated approximately 50-fold while ERα and GPR30 were down regulated, but to a much lesser degree (approximately 2-fold). ERβ was characterised as a 59kDa protein following Western blot analysis with validated antibodies and ERβ was detected in both nuclear and cytoplasmic cell compartments following immunofluorescence (IF) and immunohistochemical (IHC) analysis of cultured cells. Furthermore isoform specific antibodies detected both ERβ1 and ERβ2 in DPC cultures and in situ analysis of ERβ expression in decalcified tooth/pulp sections identified the odontoblast layer of pulp cells juxtaposed to the tooth enamel as strongly reactive for both ERβ isoforms. Finally the use of isoform specific agonists identified ERβ as the main receptor responsible for the pro-osteogenic effect of oestrogenic hormones in this tissue. Our data suggest that oestrogens stimulated osteogenic differentiation in hDPCs and that this action is mediated principally through the ERβ isoform. These findings may have important consequences for the investigation and treatment of oral and periodontal pathologies which are associated with imbalances in oestrogen concentrations and action.

Original languageEnglish
Pages (from-to)296-302
Number of pages7
JournalThe Journal of Steroid Biochemistry and Molecular Biology
Volume174
Early online date12 Oct 2017
DOIs
Publication statusPublished - Nov 2017

Fingerprint

Dental Pulp
Estrogen Receptors
Pulp
Protein Isoforms
Tissue
Tooth
Bone
Estrogens
Tooth enamel
Oral Pathology
Odontoblasts
Antibodies
Physiology
Pathology
Cell membranes
Dental Enamel
GTP-Binding Proteins
Metabolism
Bone Density
Fluorescent Antibody Technique

Keywords

  • Adult
  • Cell Differentiation
  • Cells, Cultured
  • Dental Pulp
  • Estrogen Receptor Modulators
  • Estrogen Receptor alpha
  • Estrogen Receptor beta
  • Estrogens
  • Female
  • Humans
  • Nitriles
  • Osteogenesis
  • Phenols
  • Pyrazoles
  • Receptors, Estrogen
  • Receptors, G-Protein-Coupled
  • Young Adult
  • Journal Article

Cite this

Alhodhodi, A., Alkharobi, H., Humphries, M., Alkhafaji, H., El-Gendy, R., Feichtinger, G., ... Beattie, J. (2017). Oestrogen receptor β (ERβ) regulates osteogenic differentiation of human dental pulp cells. The Journal of Steroid Biochemistry and Molecular Biology, 174, 296-302. https://doi.org/10.1016/j.jsbmb.2017.10.012

Oestrogen receptor β (ERβ) regulates osteogenic differentiation of human dental pulp cells. / Alhodhodi, Aishah; Alkharobi, Hanaa; Humphries, Matthew; Alkhafaji, Hasanain; El-Gendy, Reem; Feichtinger, Georg; Speirs, Valerie; Beattie, James.

In: The Journal of Steroid Biochemistry and Molecular Biology, Vol. 174, 11.2017, p. 296-302.

Research output: Contribution to journalArticle

Alhodhodi, Aishah ; Alkharobi, Hanaa ; Humphries, Matthew ; Alkhafaji, Hasanain ; El-Gendy, Reem ; Feichtinger, Georg ; Speirs, Valerie ; Beattie, James. / Oestrogen receptor β (ERβ) regulates osteogenic differentiation of human dental pulp cells. In: The Journal of Steroid Biochemistry and Molecular Biology. 2017 ; Vol. 174. pp. 296-302.
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abstract = "Estradiol (E2) has many important actions in the tissues of the oral cavity. Disruption of E2 metabolism or alterations in systemic E2 concentrations have been associated with compromised periodontal health. In many instances such changes occur secondarily to the well characterised effects of E2 on bone physiology -especially maintenance of bone mineral density (BMD). Despite these important epidemiological findings, little is known about the mechanism of action of E2 in oral tissues or the expression and function of oestrogen receptor (ER) isoforms in these tissues. We have isolated human dental pulp cells (hDPCs), which are able to differentiate towards an osteogenic lineage under appropriate culture conditions. We show that hDPCs express ERα, ERβ1, ERβ2 and the cell membrane associated G protein-coupled ER (GPR30). Following osteogenic differentiation of hDPCs, ERβ1 and ERβ2 were up regulated approximately 50-fold while ERα and GPR30 were down regulated, but to a much lesser degree (approximately 2-fold). ERβ was characterised as a 59kDa protein following Western blot analysis with validated antibodies and ERβ was detected in both nuclear and cytoplasmic cell compartments following immunofluorescence (IF) and immunohistochemical (IHC) analysis of cultured cells. Furthermore isoform specific antibodies detected both ERβ1 and ERβ2 in DPC cultures and in situ analysis of ERβ expression in decalcified tooth/pulp sections identified the odontoblast layer of pulp cells juxtaposed to the tooth enamel as strongly reactive for both ERβ isoforms. Finally the use of isoform specific agonists identified ERβ as the main receptor responsible for the pro-osteogenic effect of oestrogenic hormones in this tissue. Our data suggest that oestrogens stimulated osteogenic differentiation in hDPCs and that this action is mediated principally through the ERβ isoform. These findings may have important consequences for the investigation and treatment of oral and periodontal pathologies which are associated with imbalances in oestrogen concentrations and action.",
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AU - El-Gendy, Reem

AU - Feichtinger, Georg

AU - Speirs, Valerie

AU - Beattie, James

N1 - AA and HA acknowledges the Royal Embassy of Saudi Arabia − Cultural Bureau (UK) for financial support. RE acknowledges WELMEC, a Centre of Excellence in Medical Engineering funded by the Wellcome Trust and EPSRC, under grant number WT 088908/Z/09/Z for financial support. We thank Dr John Hawse (Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota) for kindly providing monoclonal anti-ERβ MC10.

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N2 - Estradiol (E2) has many important actions in the tissues of the oral cavity. Disruption of E2 metabolism or alterations in systemic E2 concentrations have been associated with compromised periodontal health. In many instances such changes occur secondarily to the well characterised effects of E2 on bone physiology -especially maintenance of bone mineral density (BMD). Despite these important epidemiological findings, little is known about the mechanism of action of E2 in oral tissues or the expression and function of oestrogen receptor (ER) isoforms in these tissues. We have isolated human dental pulp cells (hDPCs), which are able to differentiate towards an osteogenic lineage under appropriate culture conditions. We show that hDPCs express ERα, ERβ1, ERβ2 and the cell membrane associated G protein-coupled ER (GPR30). Following osteogenic differentiation of hDPCs, ERβ1 and ERβ2 were up regulated approximately 50-fold while ERα and GPR30 were down regulated, but to a much lesser degree (approximately 2-fold). ERβ was characterised as a 59kDa protein following Western blot analysis with validated antibodies and ERβ was detected in both nuclear and cytoplasmic cell compartments following immunofluorescence (IF) and immunohistochemical (IHC) analysis of cultured cells. Furthermore isoform specific antibodies detected both ERβ1 and ERβ2 in DPC cultures and in situ analysis of ERβ expression in decalcified tooth/pulp sections identified the odontoblast layer of pulp cells juxtaposed to the tooth enamel as strongly reactive for both ERβ isoforms. Finally the use of isoform specific agonists identified ERβ as the main receptor responsible for the pro-osteogenic effect of oestrogenic hormones in this tissue. Our data suggest that oestrogens stimulated osteogenic differentiation in hDPCs and that this action is mediated principally through the ERβ isoform. These findings may have important consequences for the investigation and treatment of oral and periodontal pathologies which are associated with imbalances in oestrogen concentrations and action.

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KW - Humans

KW - Nitriles

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KW - Phenols

KW - Pyrazoles

KW - Receptors, Estrogen

KW - Receptors, G-Protein-Coupled

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