title: Functional role of mechanosensitive ion channel Piezo1 in human periodontal ligament cells.
abstract: OBJECTIVE
  To evaluate the function of Piezo1, an evolutionarily conserved mechanically activated channel, in periodontal ligament (PDL) tissue homeostasis under compressive loading.
  
  
  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.
authors.affiliation: a  PhD Candidate, Department of Orthodontics, State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, West China School of Stomatology, Sichuan University, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Jin
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.forename: Yating
authors.initials: Y
authors.lastname: Wang
authors.forename: Rui
authors.initials: R
authors.lastname: Ye
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Feng
authors.forename: Zheng
authors.initials: Z
authors.lastname: Jing
authors.forename: Zhihe
authors.initials: Z
authors.lastname: Zhao
mesh_terms: D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D001696:Biomechanical Phenomena; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D051546:Cyclooxygenase 2 / Q000032:analysis; D015232:Dinoprostone / Q000032:analysis; D006706:Homeostasis / Q000502:physiology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007473:Ion Channels / Q000037:antagonists & inhibitors / Q000502:physiology*; D007527:Isoenzymes / Q000032:analysis; D051379:Mice; D009000:Monocytes / Q000187:drug effects; D016328:NF-kappa B / Q000032:analysis; D010010:Osteoclasts / Q000502:physiology; D053244:Osteoprotegerin / Q000032:analysis; D010455:Peptides / Q000494:pharmacology; D010513:Periodontal Ligament / Q000166:cytology*; D053245:RANK Ligand / Q000032:analysis; D015398:Signal Transduction / Q000502:physiology; D013111:Spider Venoms / Q000494:pharmacology; D013314:Stress, Mechanical; D000071681:Tartrate-Resistant Acid Phosphatase; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D007473:Ion Channels; D007527:Isoenzymes; C411249:MTx4 protein, Grammostola spatulata; D016328:NF-kappa B; D053244:Osteoprotegerin; C515204:PIEZO1 protein, human; D010455:Peptides; D053245:RANK Ligand; D013111:Spider Venoms; D014409:Tumor Necrosis Factor-alpha; D051546:Cyclooxygenase 2; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; D015232:Dinoprostone
keywords: Mechanical loading; NF-κB; Osteoclastogenesis; Periodontal ligament; Piezo channel