title: Regulation of OPG and RANKL expressed by human dental follicle cells in osteoclastogenesis.
abstract: We investigate whether the expression of the receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG) in human dental follicle cells (HDFCs) regulated by colony stimulating factor 1 (CSF-1), parathyroid hormone-related protein (PTHrP) and bone morphogenetic protein-2 (BMP-2) contributes to osteoclastogenesis. Adolescent human impacted third mandibular molars were used to separate HDFCs. These cells were incubated with PTHrP (10 ng/ml), CSF-1 (25 ng/ml), or BMP-2 (100 ng/ml) for 0.5, 1, 3, 6 and 12 h. The expression of OPG and RANKL was investigated by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). Two co-culture systems and tartrate-resistant acid phosphatase (TRAP) staining were used to examine osteoclast formation. Scanning electron microscopy was utilized for the resorption pit assay. RANKL and OPG were expressed innately in HDFCs. Exogenous PTHrP, CSF-1 and BMP-2 chronologically regulated the expression of RANKL and OPG in HDFCs. PTHrP and CSF-1 had similar regulative patterns leading to the up-regulated expression of RANKL and the down-regulated expression of OPG and opposite for BMP-2. The number of TRAP-positive peripheral blood mononuclear cells (PBMCs) slightly increased in contacted co-culture of HDFCs and PBMCs, whereas secreted OPG from HDFCs inhibited osteoclastogenesis in the transwell co-culture system. Contacted co-culture of HDFCs and PBMCs exhibited small and shallow resorption pits, whereas in the transwell co-culture system, secreted OPG from HDFCs reduced the resorption pits, reflecting the difference in osteoclast production. Collectively, we found a dual action of HDFCs in osteoclastogenesis; moreover, PTHrP, CSF-1 and BMP-2 might influence osteoclastogenesis by regulating the expression of RANKL and OPG in HDFCs.
authors.affiliation: Department of Stomatology, Affiliated Hospital, Academy of Military Medical Sciences, 8th Dong Da Street, Beijing, 100071, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Sun
authors.affiliation: Department of Stomatology, Affiliated Hospital, Academy of Military Medical Sciences, 8th Dong Da Street, Beijing, 100071, China.
authors.forename: Qihong
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Stomatology, 150th Hospital, PLA, Luoyang, 471000, China.
authors.forename: Yongkuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Stomotology, General Hospital of Ji'nan Military Area, Jinan, 25000, China.
authors.forename: Yingchun
authors.initials: Y
authors.lastname: Bi
authors.affiliation: Department of Stomatology, Affiliated Hospital, Academy of Military Medical Sciences, 8th Dong Da Street, Beijing, 100071, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Stomatology, Affiliated Hospital, Academy of Military Medical Sciences, 8th Dong Da Street, Beijing, 100071, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Shu
authors.affiliation: Department of Orthodontics, College of Stomatology, Zhe Jiang University, Hangzhou, 31000, China.
authors.forename: Xuepeng
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Orthodontics, Hospital of Stomatology, Fourth Military University, 145th Chang Le West Road, Xi'an, 710032, China. zuolinj@yahoo.com.
authors.forename: Zuolin
authors.initials: Z
authors.lastname: Jin
authors.affiliation: Department of Stomatology, Affiliated Hospital, Academy of Military Medical Sciences, 8th Dong Da Street, Beijing, 100071, China. drgechengde@163.com.
authors.forename: Cheng
authors.initials: C
authors.lastname: Ge
mesh_terms: D000293:Adolescent; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D003795:Dental Sac / Q000378:metabolism*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D010010:Osteoclasts / Q000378:metabolism*; D053244:Osteoprotegerin / Q000378:metabolism*; D053246:Receptor Activator of Nuclear Factor-kappa B / Q000378:metabolism*
chemical_list: D008562:Membrane Glycoproteins; D053244:Osteoprotegerin; D053246:Receptor Activator of Nuclear Factor-kappa B
keywords: Cell; Human dental follicle cells; Osteoclastogenesis; Osteoprotegrin; RANKL