title: Chondrogenesis of periodontal ligament stem cells by transforming growth factor-β3 and bone morphogenetic protein-6 in a normal healthy impacted third molar.
abstract: The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its multipotency. However, the proof of chondrogenic potential of the cells is scarce. Therefore, we investigated the chondrogenic differentiation capacity of periodontal ligament derived mesenchymal stem cells induced by transforming growth factor (TGF)-β3 and bone morphogenetic protein (BMP)-6. After isolation of periodontal ligament stem cells (PDLSCs) from human periodontal ligament, the cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 20% fetal bovine serum (FBS). A mechanical force initiated chondrogenic differentiation of the cells. For chondrogenic differentiation, 10 µg·L⁻¹ TGF-β3 or 100 µg∙L⁻¹ BMP-6 and the combination treating group for synergistic effect of the growth factors. We analyzed the PDLSCs by fluorescence-activated cell sorting and chondrogenesis were evaluated by glycosaminoglycans assay, histology, immunohistochemistry and genetic analysis. PDLSCs showed mesenchymal stem cell properties proved by FACS analysis. Glycosaminoglycans contents were increased 217% by TGF-β3 and 220% by BMP-6. The synergetic effect of TGF-β3 and BMP-6 were shown up to 281% compared to control. The combination treatment increased Sox9, aggrecan and collagen II expression compared with not only controls, but also TGF-β3 or BMP-6 single treatment dramatically. The histological analysis also indicated the chondrogenic differentiation of PDLSCs in our conditions. The results of the present study demonstrate the potential of the dental stem cell as a valuable cell source for chondrogenesis, which may be applicable for regeneration of cartilage and bone fracture in the field of cell therapy.
authors.affiliation: Laboratory of Developmental Biology and Stem Cell Differentiation/Transplantation, Department of Dental Regenerative Biotechnology, School of Dentistry, Seoul National University, Seoul, Korea.
authors.forename: Sunyoung
authors.initials: S
authors.lastname: Choi
authors.forename: Tae-Jun
authors.initials: TJ
authors.lastname: Cho
authors.forename: Soon-Keun
authors.initials: SK
authors.lastname: Kwon
authors.forename: Gene
authors.initials: G
authors.lastname: Lee
authors.forename: Jaejin
authors.initials: J
authors.lastname: Cho
mesh_terms: D053687:Adult Stem Cells / Q000502:physiology; D053674:Aggrecans / Q000032:analysis; D055418:Bone Morphogenetic Protein 6 / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation; D020219:Chondrogenesis / Q000187:drug effects / Q000502:physiology*; D024043:Collagen Type II / Q000032:analysis; D005434:Flow Cytometry; D006025:Glycosaminoglycans / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000502:physiology*; D008964:Molar, Third / Q000166:cytology*; D010513:Periodontal Ligament / Q000166:cytology* / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000032:analysis; D013314:Stress, Mechanical; D014095:Tooth, Impacted / Q000473:pathology*; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology*
chemical_list: D053674:Aggrecans; C525816:BMP6 protein, human; D055418:Bone Morphogenetic Protein 6; D024043:Collagen Type II; D006025:Glycosaminoglycans; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D053782:Transforming Growth Factor beta3