title: Expression of core-binding factor α1 and osteocalcin in fluoride-treated fibroblasts and osteoblasts.
abstract: To study the effects and importance of fluoride on FBs in the development of extraperiosteal calcification and the ossification of skeletal fluorosis, the presence of the osteogenic phenotype, which is indicated by the expression of core-binding factor α1 (Cbfa1) and osteocalcin (OCN), in an FB cell line (L929) and in osteoblasts (OBs) exposed to fluoride was determined. Fibroblasts and osteoblasts were exposed to different concentrations of fluoride (0, 0.0001, 0.001, 0.1, 1.0, 10.0 and 20.0 mg/L F(-)). By using RT-PCR and ELISA, the mRNA levels of Cbfa1 and OCN were measured at 48 h, and the protein levels of Cbfa1 and OCN were measured at 2, 4, 24, 48 and 72 h. The data demonstrated the following: (1) The Cbfa1 protein level in fluoride-treated fibroblasts clearly increased at 48 h in the groups treated with 0.0001, 0.001, 0.1, 1.0 and 20.0 mg/L F(-). The Cbfa1 protein level of the group treated with 10 mg/L F(-) at 72 h was higher than that of the control group. The level of Cbfa1 mRNA in the fibroblasts was much higher at 48 h in the group treated with 10.0 mg/L F(-) than in the control group. (2) The OCN protein level in fluoride-treated fibroblasts was significantly higher than that of the control group in the 0.0001, 0.1, 1.0, 10.0 and 20.0 mg/L F(-) groups at 2 h, and in the 0.001 and 0.1 F(-) groups at 4 h. A slightly higher level of OCN mRNA in fluoride-treated fibroblasts was also found in the 1.0 and 20.0 mg/L F(-) groups compared to the control group. (3) The expressions of Cbfa1 and OCN in osteoblasts treated with the same experimental conditions as the fibroblasts were up-regulated by fluoride following the same trend as in the fibroblasts. Our results showed an increase in the expression of Cbfa1 and OCN in fibroblasts and osteoblasts exposed to fluoride and suggested that the osteogenic function of fibroblasts induced by fluoride could play an important role in the development of extraperiosteal ossification during skeletal fluorosis.
authors.affiliation: Department of Rehabilitation Medicine of the Second Hospital Norman Bethune of Jilin University, Changchun 130041, China.
authors.forename: Xiaoqin
authors.initials: X
authors.lastname: Duan
authors.affiliation: Institute of Endemic Disease of Jilin University, 1163 Xinmin Street, Changchun 130021, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Xu
authors.affiliation: The First Hospital Norman Bethune of Jilin University, Changchun 130021, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Rehabilitation Medicine of the Second Hospital Norman Bethune of Jilin University, Changchun 130041, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Wang
authors.affiliation: Institute of Endemic Disease of Jilin University, 1163 Xinmin Street, Changchun 130021, China.
authors.forename: Guangsheng
authors.initials: G
authors.lastname: Li
authors.affiliation: Institute of Endemic Disease of Jilin University, 1163 Xinmin Street, Changchun 130021, China. Electronic address: jingling85619287@163.com.
authors.forename: Ling
authors.initials: L
authors.lastname: Jing
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005459:Fluorides / Q000494:pharmacology; D007150:Immunohistochemistry; D051379:Mice; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism*; D015675:Osteocalcin / Q000378:metabolism*
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D015675:Osteocalcin; D005459:Fluorides
keywords: Core-binding factor α1 (Cbfa1); Fibroblast (FB); Fluoride; Osteoblast (OB); Osteocalcin (OCN)