RANKL Expression Is Related to the Differentiation State of Human Osteoblasts

Gerald J. Atkins, Panagiota Kostakis, Beiqing Pan, Amanda Farrugia, Stan Gronthos, Andreas Evdokiou, Kate Harrison, David M. Findlay, Andrew C W Zannettino

Research output: Contribution to journalArticle

171 Citations (Scopus)

Abstract

Human osteoblast phenotypes that support osteoclast differentiation and bone formation are not well characterized. Osteoblast differentiation markers were examined in relation to RANKL expression. RANKL expression was induced preferentially in immature cells. These results support an important link between diverse osteoblast functions. Cells of the osteoblast lineage support two apparently distinct functions: bone formation and promotion of osteoclast formation. The aim of this study was to examine the relationship between these phenotypes in human osteoblasts (NHBC), in terms of the pre-osteoblast marker, STRO-1, and the mature osteoblast marker, alkaline phosphatase (AP), and the expression of genes involved in osteoclast formation, RANKL and OPG. The osteotropic stimuli, 1α,25(OH)2vitamin D3 (vitD3) and dexamethasone, were found to have profound proliferative and phenotypic effects on NHBCs. VitD3 inhibited NHBC proliferation and increased the percentage of cells expressing STRO-1 over an extended culture period, implying that vitD3 promotes and maintains an immature osteogenic phenotype. Concomitantly, RANKL mRNA expression was upregulated and maintained in NHBC in response to vitD3. Dexamethasone progressively promoted the proliferation of AP-expressing cells, resulting in the overall maturation of the cultures. Dexamethasone had little effect on RANKL mRNA expression and downregulated OPG mRNA expression in a donor-dependent manner. Regression analysis showed that RANKL mRNA expression was associated negatively with the percentage of cells expressing AP (p < 0.01) in vitD3- and dexamethasone-treated NHBCs. In contrast, RANKL mRNA expression was associated positively with the percentage of STRO-1+ cells (p < 0.01). In NHBCs sorted by FACS based on STRO-1 expression (STRO-1bright and STRO-1dim populations), it was found that vitD3 upregulated the expression of RANKL mRNA preferentially in STRO-1bright cells. The results suggest that immature osteoblasts respond to osteotropic factors in a potentially pro-osteoclastogenic manner. Additionally, the dual roles of osteoblasts, in supporting osteoclastogenesis or forming bone, may be performed by the same lineage of cells at different stages of their maturation.

LanguageEnglish
Pages1088-1098
Number of pages11
JournalJournal of Bone and Mineral Research
Volume18
Issue number6
DOIs
Publication statusPublished - 1 Jun 2003
Externally publishedYes

Keywords

  • Glucocorticoid
  • Osteoblast
  • RANKL
  • STRO-1
  • Vitamin D

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Orthopedics and Sports Medicine

Cite this

Atkins, Gerald J. ; Kostakis, Panagiota ; Pan, Beiqing ; Farrugia, Amanda ; Gronthos, Stan ; Evdokiou, Andreas ; Harrison, Kate ; Findlay, David M. ; Zannettino, Andrew C W. / RANKL Expression Is Related to the Differentiation State of Human Osteoblasts. In: Journal of Bone and Mineral Research. 2003 ; Vol. 18, No. 6. pp. 1088-1098.
@article{70c431df43984a9db99acf0cbe0fb2bc,
title = "RANKL Expression Is Related to the Differentiation State of Human Osteoblasts",
abstract = "Human osteoblast phenotypes that support osteoclast differentiation and bone formation are not well characterized. Osteoblast differentiation markers were examined in relation to RANKL expression. RANKL expression was induced preferentially in immature cells. These results support an important link between diverse osteoblast functions. Cells of the osteoblast lineage support two apparently distinct functions: bone formation and promotion of osteoclast formation. The aim of this study was to examine the relationship between these phenotypes in human osteoblasts (NHBC), in terms of the pre-osteoblast marker, STRO-1, and the mature osteoblast marker, alkaline phosphatase (AP), and the expression of genes involved in osteoclast formation, RANKL and OPG. The osteotropic stimuli, 1α,25(OH)2vitamin D3 (vitD3) and dexamethasone, were found to have profound proliferative and phenotypic effects on NHBCs. VitD3 inhibited NHBC proliferation and increased the percentage of cells expressing STRO-1 over an extended culture period, implying that vitD3 promotes and maintains an immature osteogenic phenotype. Concomitantly, RANKL mRNA expression was upregulated and maintained in NHBC in response to vitD3. Dexamethasone progressively promoted the proliferation of AP-expressing cells, resulting in the overall maturation of the cultures. Dexamethasone had little effect on RANKL mRNA expression and downregulated OPG mRNA expression in a donor-dependent manner. Regression analysis showed that RANKL mRNA expression was associated negatively with the percentage of cells expressing AP (p < 0.01) in vitD3- and dexamethasone-treated NHBCs. In contrast, RANKL mRNA expression was associated positively with the percentage of STRO-1+ cells (p < 0.01). In NHBCs sorted by FACS based on STRO-1 expression (STRO-1bright and STRO-1dim populations), it was found that vitD3 upregulated the expression of RANKL mRNA preferentially in STRO-1bright cells. The results suggest that immature osteoblasts respond to osteotropic factors in a potentially pro-osteoclastogenic manner. Additionally, the dual roles of osteoblasts, in supporting osteoclastogenesis or forming bone, may be performed by the same lineage of cells at different stages of their maturation.",
keywords = "Glucocorticoid, Osteoblast, RANKL, STRO-1, Vitamin D",
author = "Atkins, {Gerald J.} and Panagiota Kostakis and Beiqing Pan and Amanda Farrugia and Stan Gronthos and Andreas Evdokiou and Kate Harrison and Findlay, {David M.} and Zannettino, {Andrew C W}",
year = "2003",
month = "6",
day = "1",
doi = "10.1359/jbmr.2003.18.6.1088",
language = "English",
volume = "18",
pages = "1088--1098",
journal = "Journal of Bone and Mineral Research",
issn = "0884-0431",
publisher = "Wiley-Blackwell",
number = "6",

}

Atkins, GJ, Kostakis, P, Pan, B, Farrugia, A, Gronthos, S, Evdokiou, A, Harrison, K, Findlay, DM & Zannettino, ACW 2003, 'RANKL Expression Is Related to the Differentiation State of Human Osteoblasts', Journal of Bone and Mineral Research, vol. 18, no. 6, pp. 1088-1098. https://doi.org/10.1359/jbmr.2003.18.6.1088

RANKL Expression Is Related to the Differentiation State of Human Osteoblasts. / Atkins, Gerald J.; Kostakis, Panagiota; Pan, Beiqing; Farrugia, Amanda; Gronthos, Stan; Evdokiou, Andreas; Harrison, Kate; Findlay, David M.; Zannettino, Andrew C W.

In: Journal of Bone and Mineral Research, Vol. 18, No. 6, 01.06.2003, p. 1088-1098.

Research output: Contribution to journalArticle

TY - JOUR

T1 - RANKL Expression Is Related to the Differentiation State of Human Osteoblasts

AU - Atkins, Gerald J.

AU - Kostakis, Panagiota

AU - Pan, Beiqing

AU - Farrugia, Amanda

AU - Gronthos, Stan

AU - Evdokiou, Andreas

AU - Harrison, Kate

AU - Findlay, David M.

AU - Zannettino, Andrew C W

PY - 2003/6/1

Y1 - 2003/6/1

N2 - Human osteoblast phenotypes that support osteoclast differentiation and bone formation are not well characterized. Osteoblast differentiation markers were examined in relation to RANKL expression. RANKL expression was induced preferentially in immature cells. These results support an important link between diverse osteoblast functions. Cells of the osteoblast lineage support two apparently distinct functions: bone formation and promotion of osteoclast formation. The aim of this study was to examine the relationship between these phenotypes in human osteoblasts (NHBC), in terms of the pre-osteoblast marker, STRO-1, and the mature osteoblast marker, alkaline phosphatase (AP), and the expression of genes involved in osteoclast formation, RANKL and OPG. The osteotropic stimuli, 1α,25(OH)2vitamin D3 (vitD3) and dexamethasone, were found to have profound proliferative and phenotypic effects on NHBCs. VitD3 inhibited NHBC proliferation and increased the percentage of cells expressing STRO-1 over an extended culture period, implying that vitD3 promotes and maintains an immature osteogenic phenotype. Concomitantly, RANKL mRNA expression was upregulated and maintained in NHBC in response to vitD3. Dexamethasone progressively promoted the proliferation of AP-expressing cells, resulting in the overall maturation of the cultures. Dexamethasone had little effect on RANKL mRNA expression and downregulated OPG mRNA expression in a donor-dependent manner. Regression analysis showed that RANKL mRNA expression was associated negatively with the percentage of cells expressing AP (p < 0.01) in vitD3- and dexamethasone-treated NHBCs. In contrast, RANKL mRNA expression was associated positively with the percentage of STRO-1+ cells (p < 0.01). In NHBCs sorted by FACS based on STRO-1 expression (STRO-1bright and STRO-1dim populations), it was found that vitD3 upregulated the expression of RANKL mRNA preferentially in STRO-1bright cells. The results suggest that immature osteoblasts respond to osteotropic factors in a potentially pro-osteoclastogenic manner. Additionally, the dual roles of osteoblasts, in supporting osteoclastogenesis or forming bone, may be performed by the same lineage of cells at different stages of their maturation.

AB - Human osteoblast phenotypes that support osteoclast differentiation and bone formation are not well characterized. Osteoblast differentiation markers were examined in relation to RANKL expression. RANKL expression was induced preferentially in immature cells. These results support an important link between diverse osteoblast functions. Cells of the osteoblast lineage support two apparently distinct functions: bone formation and promotion of osteoclast formation. The aim of this study was to examine the relationship between these phenotypes in human osteoblasts (NHBC), in terms of the pre-osteoblast marker, STRO-1, and the mature osteoblast marker, alkaline phosphatase (AP), and the expression of genes involved in osteoclast formation, RANKL and OPG. The osteotropic stimuli, 1α,25(OH)2vitamin D3 (vitD3) and dexamethasone, were found to have profound proliferative and phenotypic effects on NHBCs. VitD3 inhibited NHBC proliferation and increased the percentage of cells expressing STRO-1 over an extended culture period, implying that vitD3 promotes and maintains an immature osteogenic phenotype. Concomitantly, RANKL mRNA expression was upregulated and maintained in NHBC in response to vitD3. Dexamethasone progressively promoted the proliferation of AP-expressing cells, resulting in the overall maturation of the cultures. Dexamethasone had little effect on RANKL mRNA expression and downregulated OPG mRNA expression in a donor-dependent manner. Regression analysis showed that RANKL mRNA expression was associated negatively with the percentage of cells expressing AP (p < 0.01) in vitD3- and dexamethasone-treated NHBCs. In contrast, RANKL mRNA expression was associated positively with the percentage of STRO-1+ cells (p < 0.01). In NHBCs sorted by FACS based on STRO-1 expression (STRO-1bright and STRO-1dim populations), it was found that vitD3 upregulated the expression of RANKL mRNA preferentially in STRO-1bright cells. The results suggest that immature osteoblasts respond to osteotropic factors in a potentially pro-osteoclastogenic manner. Additionally, the dual roles of osteoblasts, in supporting osteoclastogenesis or forming bone, may be performed by the same lineage of cells at different stages of their maturation.

KW - Glucocorticoid

KW - Osteoblast

KW - RANKL

KW - STRO-1

KW - Vitamin D

UR - http://www.scopus.com/inward/record.url?scp=0142059624&partnerID=8YFLogxK

U2 - 10.1359/jbmr.2003.18.6.1088

DO - 10.1359/jbmr.2003.18.6.1088

M3 - Article

VL - 18

SP - 1088

EP - 1098

JO - Journal of Bone and Mineral Research

T2 - Journal of Bone and Mineral Research

JF - Journal of Bone and Mineral Research

SN - 0884-0431

IS - 6

ER -