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).
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).
Authors: Mervi Gürsoy; Fares Zeidán-Chuliá; Eija Könönen; José C F Moreira; Joonas Liukkonen; Timo Sorsa; Ulvi K Gürsoy Journal: OMICS Date: 2014-07-01
Authors: Ling-Ling E; Wen-Huan Xu; Lin Feng; Yi Liu; Dong-Qing Cai; Ning Wen; Wen-Jie Zheng Journal: Int J Mol Med Date: 2016-04-12 Impact factor: 4.101