J Korean Assoc Oral Maxillofac Surg.  2014 Dec;40(6):291-296. 10.5125/jkaoms.2014.40.6.291.

Effect of recombinant human bone morphogenetic protein-2 on bisphosphonate-treated osteoblasts

• Kwon TK
• Song JM
• Kim IR
• Park BS
• Kim CH
• Cheong IK
• Shin SH

• KMID: 1799563
• DOI: http://doi.org/10.5125/jkaoms.2014.40.6.291

Abstract

OBJECTIVES
Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is a side effect of bisphophonate therapy that has been reported in recent years. Osteoclastic inactivity by bisphosphonate is the known cause of BRONJ. Bone morphogenetic protein-2 (BMP-2) plays an important role in the development of bone. Recombinant human BMP-2 (rhBMP-2) is potentially useful as an activation factor for bone repair. We hypothesized that rhBMP-2 would enhance the osteoclast-osteoblast interaction related to bone remodeling.
MATERIALS AND METHODS
Human fetal osteoblast cells (hFOB 1.19) were treated with 100 microM alendronate, and 100 ng/mL rhBMP-2 was added. Cells were incubated for a further 48 hours, and cell viability was measured using an MTT assay. Expression of the three cytokines from osteoblasts, receptor activator of nuclear factor-kappaB ligand (RANKL), osteoprotegerin (OPG), and macrophage colony-stimulating factor (M-CSF), were analyzed by real-time polymerase chain reaction and enzyme-linked immunosorbent assay.
RESULTS
Cell viability was decreased to 82.75%+/-1.00% by alendronate and then increased to 110.43%+/-1.35% after treatment with rhBMP-2 (P<0.05, respectively). OPG, RANKL, and M-CSF expression were all decreased by alendronate treatment. RANKL and M-CSF expression were increased, but OPG was not significantly affected by rhBMP-2.
CONCLUSION
rhBMP2 does not affect OPG gene expression in hFOB, but it may increase RANKL and M-CSF gene expression.

Keyword

Bone morphogenetic protein-2; Alendronate; Osteoblasts; Macrophage colony-stimulating factor; RANK ligand

MeSH Terms

Alendronate
Bisphosphonate-Associated Osteonecrosis of the Jaw
Bone Remodeling
Cell Survival
Cytokines
Enzyme-Linked Immunosorbent Assay
Gene Expression
Humans
Macrophage Colony-Stimulating Factor
Osteoblasts*
Osteoclasts
Osteoprotegerin
RANK Ligand
Real-Time Polymerase Chain Reaction
Alendronate
Cytokines
Macrophage Colony-Stimulating Factor
Osteoprotegerin
RANK Ligand

Figure

• 

  Fig. 1 Cell viability of various alendronate concentrations in hFOB 1.19.

• 

  Fig. 2 Effect of alendronate and rhBMP-2 on OPG expression. ELISA of OPG expression. (rhBMP-2: recombinant human bone morphogenetic protein-2, OPG: osteoprotegerin, ELISA: enzymelinked immunosorbent assay)

• 

  Fig. 3 Effect of alendronate and rhBMP-2 on RANKL expression. ELISA of RANKL expression. (rhBMP-2: recombinant human bone morphogenetic protein-2, RANKL: receptor activator of nuclear factor-κB ligand, ELISA: enzyme-linked immunosorbent assay)

• 

  Fig. 4 Effect of alendronate and rhBMP-2 on M-CSF expression. ELISA of M-CSF expression. (rhBMP-2: recombinant human bone morphogenetic protein-2, M-CSF: macrophage colony-stimulating factor, ELISA: enzyme-linked immunosorbent assay)

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