title: Effects ofrhBMP-2gene transfection to periodontal ligament cells on osteogenesis.
abstract: The present study aims to investigate the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on the osteogenesis of periodontal ligament (PDL) cells. The expression vector of rhBMP-2 (pcDNA3.1-rhBMP-2) was established. PDL cells were obtained through the enzymatic digestion and tissue explant methods and verified by immunohistochemistry. Cells were classified into experimental (cells transfected with pcDNA3.1/rhBMP-2-EGFP), blank (cells with no transfection) and control group (cells transfected with empty plasmid). rhBMP-2 expression was assessed via Western blotting analysis. The mineralization ability, alkaline phosphatase (ALP) activity and level of related osteogenic biomarkers were detected to evaluate the osteogenic characteristics of PDL cells. The rhBMP-2 expression vector (pcDNA3.1-rhBMP-2) was successfully established. Primary PDL cells displayed a star or long, spindle shape. The cultured cells were long, spindle-shaped, had a plump cell body and homogeneous cytoplasm and the ellipse nucleus contained two or three nucleoli. Cells displayed a radial, sheaf-like or eddy-like arrangement after adherence growth. Immunohistochemical staining confirmed that cells originated from mesenchymal opposed to epithelium. The experimental group exhibited an enhanced mineralization ability, higher ALP activity and increased expression of rhBMP-2 and osteogenic biomarkers (Runx2, collagen type I and osteocalcin) than the blank and control group. The present study demonstrated that rhBMP-2 transfection enhances the osteogenesis of PDL cells and provides a possibility for the application of rhBMP-2 expression products in dental disease treatment.
authors.affiliation: Department of Stomatology, PLA General Hospital of Chengdu Military Region, Chengdu 610083, China.
authors.forename: Cong-Xiang
authors.initials: CX
authors.lastname: Jian
authors.affiliation: Chengdu Military Garrison Center for Disease Control and Prevention, Chengdu 650032, China.
authors.forename: Quan-Shui
authors.initials: QS
authors.lastname: Fan
authors.affiliation: Chengdu Military Garrison Center for Disease Control and Prevention, Chengdu 650032, China.
authors.forename: Yong-He
authors.initials: YH
authors.lastname: Hu
authors.affiliation: Department of Stomatology, PLA General Hospital of Chengdu Military Region, Chengdu 610083, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: He
authors.affiliation: Department of Stomatology, PLA General Hospital of Chengdu Military Region, Chengdu 610083, China.
authors.forename: Ming-Zhe
authors.initials: MZ
authors.lastname: Li
authors.affiliation: Department of Stomatology, PLA General Hospital of Chengdu Military Region, Chengdu 610083, China.
authors.forename: Wei-Yin
authors.initials: WY
authors.lastname: Zheng
authors.affiliation: Department of Stomatology, PLA General Hospital of Chengdu Military Region, Chengdu 610083, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Ren
authors.affiliation: Department of Stomatology, PLA General Hospital of Chengdu Military Region, Chengdu 610083, China chenjunlili2016@163.com.
authors.forename: Chen-Jun
authors.initials: CJ
authors.lastname: Li
mesh_terms: D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D002467:Cell Nucleus / Q000235:genetics; D024042:Collagen Type I / Q000378:metabolism; D003593:Cytoplasm / Q000235:genetics; D004848:Epithelium / Q000378:metabolism; D006801:Humans; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000235:genetics*; D010513:Periodontal Ligament / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection / Q000379:methods; D016212:Transforming Growth Factor beta / Q000235:genetics*; D055815:Young Adult
chemical_list: D015415:Biomarkers; D055396:Bone Morphogenetic Protein 2; D024042:Collagen Type I; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; D015675:Osteocalcin
keywords: Collagen type I; Gene transfection; Osteogenesis; Periodontal ligament cells; Recombinant human bone morphogenetic protein-2; Runx2