Low-Intensity Pulsed Ultrasound Induced Osteoblast Differentiation Mediated by the PYK2-ERK2 Signaling in MC3T3-E1 Cells.

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  • Utsu Akihisa
    Division of Oral Structural and Functional Biology, Nihon University Graduate School of Dentistry Department of Orthodontics, Nihon University School of Dentistry
  • Tanabe Natsuko
    Department of Biochemistry, Nihon University School of Dentistry Division of Functional Morphology, Dental Research Center, Nihon University School of Dentistry
  • Manaka Soichiro
    Division of Functional Morphology, Dental Research Center, Nihon University School of Dentistry Department of Periodontology, Nihon University School of Dentistry
  • Tomita Keiko
    Department of Periodontology, Nihon University School of Dentistry
  • Ichikawa Risa
    Department of Periodontology, Nihon University School of Dentistry Division of Applied Oral Sciences, Nihon University Graduate School of Dentistry
  • Ono Misae
    Department of Periodontology, Nihon University School of Dentistry Division of Applied Oral Sciences, Nihon University Graduate School of Dentistry
  • Masai Yuma
    Department of Periodontology, Nihon University School of Dentistry Division of Applied Oral Sciences, Nihon University Graduate School of Dentistry
  • Suzuki Naoto
    Department of Biochemistry, Nihon University School of Dentistry Division of Functional Morphology, Dental Research Center, Nihon University School of Dentistry
  • Motoyoshi Mitsuru
    Department of Orthodontics, Nihon University School of Dentistry Division of Clinical Research, Dental Research Center, Nihon University School of Dentistry

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<p>Low-intensity pulsed ultrasonic (LIPUS) is a noninvasive force promoting bone regeneration as mechanical stimulation. Integrins mediate numerous intracellular signaling pathways, including mitogen-activated protein kinase (MAPK) and calcium influx through focal adhesion. A non-receptor tyrosine kinase of proline-rich tyrosine kinase 2 (PYK2) associates with integrins at focal adhesions. A previous study reported that intracellular Ca2+ levels regulate the activation of PYK2 induced by mechanical strain in osteoblasts. However, the roles of PYK2 on the expression of extracellular matrix (ECM) proteins involved in osteogenesis and osteoblast differentiation induced by LIPUS remain unclear. We aimed to investigate the roles of PYK2 on the expression of transcription factors runt-related transcription factor 2 (Runx2) and osterix, which regulate osteoblast differentiation, ECM proteins, and the downstream intracellular signaling pathway, including PYK2 and focal adhesion kinase (FAK) in MC3T3-E1 cells when treated with LIPUS. LIPUS enhanced the phosphorylation of PYK2 and extracellular signal-regulated kinase2 (ERK2) and induced the expression of Runx2, osterix, type I collagen, osteocalcin, and calcium content in ECM. However, siPYK2 and PYK2 inhibitor PF431396 suppressed these stimulatory effects of LIPUS. These results suggest that LIPUS induces osteoblast differentiation mediated by the PYK2-ERK2 signaling in MC3T3-E1 cells.</p>

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