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Horm Metab Res 2008; 40(12): 869-874
DOI: 10.1055/s-0028-1082083
Original Basic

© Georg Thieme Verlag KG Stuttgart · New York

Modifying RANKL/OPG mRNA Expression in Differentiating and Growing Human Primary Osteoblasts

M. Giner 1 , M. J. Montoya 2 , M. A. Vázquez 2 , M. J. Rios 2 , R. Moruno 2 , M. J. Miranda 1 , R. Pérez-Cano 1 , 2
  • 1Bone Metabolism Unit, Internal Medicine, University Hospital “Virgen Macarena” Seville, Spain
  • 2Medicine Department, University of Seville, Spain
Further Information

Publication History

received 29.10.2007

accepted 19.05.2008

Publication Date:
17 October 2008 (online)

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Abstract

The OPG/RANKL system in primary cultures of human osteoblasts has been studied by different authors. However, very few studies have been performed on gene expression of RANKL and OPG at different stages of maturation on human osteoblast cultures. The effect of 17-β-estradiol and 1,25dihydroxyvitamin D3 on the OPG/RANKL system is not known during the different states of cellular maturation. In this work we quantified OPG and RANKL protein levels (ELISA) and the mRNA of OPG, RANKL, collagen type I, alkaline phosphatase, and osteocalcin (semi-quantitative RT-PCR) in human osteoblasts. We analyzed these in basal conditions and after incubation with 17-β-estradiol and 1,25dihydroxyvitamin D3 in the first and second phases. We found that OPG secretion and expression levels increased throughout cellular growth. RANKL proteins were detected only in the first stage, and the expression increased throughout the first phase. Thus, the RANKL/OPG ratio was higher in immature osteoblasts than in mature osteoblasts. The evolution of RANKL gene expression was related to collagen I and alkaline phosphatase, while OPG was related to osteocalcin. We observed no modifications after estradiol and 1,25dihydroxyvitamin D3 treatment. Our results suggest that the OB is a positive stimulator at precocious stages of differentiation on osteoclastogenic modulates.

Key words

human osteoblast culture - 17-β-estradiol - 1,25dihydroxyvitamin D3 - osteoporosis - osteoarthritis

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Correspondence

M. Giner

Bone Metabolism Unit

Internal Medicine

University Hospital

Av. Dr. Fedriani s/n

41018 Seville

Spain

Phone: +34/954/55 66 16 17

Fax: +34/954/55 66 17

Email: merce_giner@yahoo.es