Serial administration of rhBMP-2 and alendronate enhances the differentiation of osteoblasts

Int J Rheum Dis. 2021 Oct;24(10):1266-1272. doi: 10.1111/1756-185X.14189. Epub 2021 Jul 29.

Abstract

Aim: The incorporation of growth factors is an effective strategy to accelerate bone induction. Bone morphogenetic protein-2 (BMP-2) promotes osteoblast differentiation and induces bone formation. Alendronate (ALN) is an osteoclast deactivation drug. We investigated the effect of serial administration of recombinant human BMP-2 (rhBMP-2) and ALN on osteoblast differentiation.

Methods: The effect of serial administration of rhBMP-2 (0-150 ng/mL) and ALN (0-15 µmol/L) on the viability and differentiation of a clonal murine calvarial cell line, MC3T3-E1, was evaluated at various concentrations and for different periods. The Cell Counting Kit-8 assay was used to assess cell viability. The alkaline phosphatase activity was evaluated as an indicator of osteogenic differentiation. The expression levels of runt domain-containing transcription factor 2 (Runx2) and osteopontin (OPN) were analyzed by real-time polymerase chain reaction and western blotting. Statistical analyses were performed using Student's t test.

Results: The serial treatment with rhBMP-2 and ALN increased the expression of the differentiation-related factors Runx2 and OPN, as well as the differentiation ability of osteoblasts compared with individual or simultaneous treatment. The osteoblasts treated with rhBMP-2 followed by ALN showed the highest differentiation. The degree of differentiation in the group treated with rhBMP-2 for 7 days followed by ALN for 3 days was increased by 1.5 times compared with that of the group treated with rhBMP-2 alone (P < .01).

Conclusion: These findings indicate that the serial administration of rhBMP-2 and ALN may exert osteogenic effects on osteoblastic cells via the upregulation of Runx2 and OPN.

Keywords: alendronate; bone morphogenetic protein-2; osteogenesis.

MeSH terms

  • 3T3 Cells
  • Alendronate / pharmacology*
  • Alkaline Phosphatase / metabolism
  • Animals
  • Bone Density Conservation Agents / pharmacology*
  • Bone Morphogenetic Protein 2 / pharmacology*
  • Cell Differentiation / drug effects*
  • Core Binding Factor Alpha 1 Subunit / genetics
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Mice
  • Osteoblasts / drug effects*
  • Osteoblasts / metabolism
  • Osteogenesis / drug effects*
  • Osteopontin / genetics
  • Osteopontin / metabolism
  • Recombinant Proteins / pharmacology

Substances

  • BMP2 protein, human
  • Bone Density Conservation Agents
  • Bone Morphogenetic Protein 2
  • Core Binding Factor Alpha 1 Subunit
  • Recombinant Proteins
  • Runx2 protein, mouse
  • Spp1 protein, mouse
  • Osteopontin
  • Alkaline Phosphatase
  • Alendronate