Combined Treatment with Low-Level Laser and rhBMP-2 Promotes Differentiation and Mineralization of Osteoblastic Cells under Hypoxic Stress
Tissue Engineering and Regenerative Medicine
;
(6): 793-801, 2018.
Article
in English
| WPRIM
| ID: wpr-718786
ABSTRACT
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-1α), 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 hypoxiccultured MC3T3-E1 osteoblasts by activating p38/PKD signaling in vitro.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Osteoblasts
/
Oxygen
/
Phosphorylation
/
Phosphotransferases
/
Transcription Factors
/
In Vitro Techniques
/
Blotting, Western
/
Cell Culture Techniques
/
Collagen Type I
/
Low-Level Light Therapy
Limits:
Humans
Language:
English
Journal:
Tissue Engineering and Regenerative Medicine
Year:
2018
Type:
Article
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