The Angle orthodontist · 2015
Functional role of mechanosensitive ion channel Piezo1 in human periodontal ligament cells.
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Jin Ying, Li Juan, Wang Yating, Ye Rui, Feng Xiaoxia, Jing Zheng, Zhao Zhihe
Аннотация
OBJECTIVE: To evaluate the function of Piezo1, an evolutionarily conserved mechanically activated channel, in periodontal ligament (PDL) tissue homeostasis under compressive loading.
MATERIALS AND METHODS: Primary human PDL cells (hPDLCs) were isolated, cultured, and then subjected to 2.0 g/cm(2) static compressive loading for 0.5, 3, 6, and 12 hours, respectively. The expressions of Piezo1 and osteoclastogenesis marker gene were assessed by semiquantitative reverse transcription-polymerase chain reaction. In addition, Piezo1 inhibitor, GsMTx4, was used to block the function of Piezo1, and tumor necrosis factor-α was also used as a positive control. After 12 hours of compressive loading the PDLCs were co-cultured with murine monocytic cell line RAW264.7. Immunofluorescence, western blot, enzyme-linked immunosorbent assay, and tartrate-resistant acid phosphatase staining were also used to test the potency of PDLCs to induce osteoclastogenesis and the activation of nuclear factor (NF)-κB.
RESULTS: Piezo1, cyclooxygenase-2, receptor activator of NF-κB ligand, and prostaglandin E2 were significantly upregulated under static compressive stimuli. GsMTx4 repressed osteoclastogenesis in the mechanical stress-pretreated PDLCs-RAW264.7 co-culture system. Furthermore, NF-κB signaling pathway was involved in the mechanical stress-induced osteoclastogenesis.
CONCLUSIONS: Piezo1 exerts a transduction role in mechanical stress-induced osteoclastogenesis in hPDLCs.
MATERIALS AND METHODS: Primary human PDL cells (hPDLCs) were isolated, cultured, and then subjected to 2.0 g/cm(2) static compressive loading for 0.5, 3, 6, and 12 hours, respectively. The expressions of Piezo1 and osteoclastogenesis marker gene were assessed by semiquantitative reverse transcription-polymerase chain reaction. In addition, Piezo1 inhibitor, GsMTx4, was used to block the function of Piezo1, and tumor necrosis factor-α was also used as a positive control. After 12 hours of compressive loading the PDLCs were co-cultured with murine monocytic cell line RAW264.7. Immunofluorescence, western blot, enzyme-linked immunosorbent assay, and tartrate-resistant acid phosphatase staining were also used to test the potency of PDLCs to induce osteoclastogenesis and the activation of nuclear factor (NF)-κB.
RESULTS: Piezo1, cyclooxygenase-2, receptor activator of NF-κB ligand, and prostaglandin E2 were significantly upregulated under static compressive stimuli. GsMTx4 repressed osteoclastogenesis in the mechanical stress-pretreated PDLCs-RAW264.7 co-culture system. Furthermore, NF-κB signaling pathway was involved in the mechanical stress-induced osteoclastogenesis.
CONCLUSIONS: Piezo1 exerts a transduction role in mechanical stress-induced osteoclastogenesis in hPDLCs.
Предметные рубрики MeSH
Acid PhosphataseAnimalsBiomechanical PhenomenaCell DifferentiationCell LineCells, CulturedCoculture TechniquesCyclooxygenase 2DinoprostoneHomeostasisHumansIntercellular Signaling Peptides and ProteinsIon ChannelsIsoenzymesMiceMonocytesNF-kappa BOsteoclastsOsteoprotegerinPeptidesPeriodontal LigamentRANK LigandSignal TransductionSpider VenomsStress, MechanicalTartrate-Resistant Acid PhosphataseTime FactorsTumor Necrosis Factor-alpha