Tissue Engineering and Regenerative Medicine

  • 제15권6호
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  • Pages.793-801
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  • 2018
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  • 1738-2696(pISSN)
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  • 2212-5469(eISSN)
한국조직공학·재생의학회 (Korean Tissue Engineering and Regenerative Medicine Society)
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Combined Treatment with Low-Level Laser and rhBMP-2 Promotes Differentiation and Mineralization of Osteoblastic Cells under Hypoxic Stress

  • Heo, Jin-Ho (Department of Oral and Maxillofacial Surgery, Pusan National University) ;
  • Choi, Jeong-Hun (Department of Oral and Maxillofacial Surgery, Pusan National University) ;
  • Kim, In-Ryoung (Department of Oral Anatomy, Pusan National University) ;
  • Park, Bong-Soo (Department of Oral Anatomy, Pusan National University) ;
  • Kim, Yong-Deok (Department of Oral and Maxillofacial Surgery, Pusan National University)
  • 투고 : 2018.08.29
  • 심사 : 2018.10.31
  • 발행 : 2018.12.01
⟨ 이전 논문 다음 논문 ⟩

초록

BACKGROUND: The aim of this study was to evaluate the combined effect of low-level laser treatment (LLLT) and recombinant human bone morphological protein-2 (rhBMP-2) applied to hypoxic-cultured MC3T3-E1 osteoblastic cells and to determine possible signaling pathways underlying differentiation and mineralization of osteoblasts under hypoxia. METHODS: MC3T3-E1 cells were cultured under 1% oxygen tension for 72 h. Cell cultures were divided into four groups: normoxia control, low-level laser (LLL) alone, rhBMP-2 combined with LLLT, and rhBMP-2 under hypoxia. Laser irradiation was applied at 0, 24, and 48 h. Cells were treated with rhBMP-2 at 50 ng/mL. Alkaline phosphatase activity was measured at 3, 7, and 14 days to evaluate osteoblastic differentiation. Cell mineralization was determined with Alizarin red S staining at 7 and 14 days. Western blot assays were performed to evaluate whether p38/protein kinase D (PKD) signaling was involved. RESULTS: The results indicate that LLLT and rhBMP-2 synergistically increased alkaline phosphatase (ALP) activity and mineralization. Western blot analyses showed that expression of type I collagen, runt-related transcription factor 2 (RUNX2), and Osterix (Osx), increased and expression of hypoxia-inducible factor 1-alpha ($HIF-1{\alpha}$), decreased more in the LLLT and rhBMP-2 combined group than in the rhBMP-2 or LLL alone groups. Moreover, LLLT and rhBMP-2 stimulated p38 phosphorylation and rhBMP-2 and LLLT increased Prkd1 phosphorylation. CONCLUSION: Combined treatment with rhBMP-2 and LLL induced differentiation and mineralization of hypoxic-cultured MC3T3-E1 osteoblasts by activating p38/PKD signaling in vitro.

키워드

과제정보

연구 과제 주관 기관 : National Research Foundation of Korea, Korea Health Industry Development Institute (KHIDI)

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