Oestrogen regulates proliferation, osteoblastic differentiation, collagen synthesis and periostin gene expression in human periodontal ligament cells through oestrogen receptor beta.

Objective: The present study was designed to examine how oestrogen regulates proliferation, osteoblastic differentiation, collagen synthesis and periostin gene expression in primary human periodontal ligament (hPDL) cells.

Design: The short interfering RNA (siRNA) technique was used to inhibit oestrogen receptor beta (ERβ) expression hPDL cells. hPDL cell were isolated and fully characterized. A colorimetric assay was applied for the determination of alkaline phosphatase (ALP). An ELISA kit was used to detect osteocalcin (OCN) levels. Collagen synthesis was determined by measuring the incorporation of L-[3H] praline. RT-PCR was performed to detection of periostin mRNA relative gene expression.

Results: ERβ mRNA was expressed in hPDL cells and significant inhibition of mRNA expression and ERβ mature protein of the ERβ was evident in the siRNA group. At 72h, there was a significant increase in non-transfected hPDL cell proliferation after estradiol stimulation. Addition of 17β-estradiol significantly enhanced ALP activity and production of OCN in non-transfected cells but had no effect on collagen synthesis. A clear increase in periostin mRNA expression levels was observed after incubating hPDL cells with estradiol. In hPDL-siERβ cells, the application of estradiol did not produce any evident differences in periostin mRNA expression

Conclusions: ERβ may play important roles in oestrogen-induced effects on hPDL cell proliferation, osteoblastic differentiation and expression of key molecules for the functional and structural integrity of the periodontium (i.e. periostin).