Porcine Bone Incorporated With 4-Hexylresorcinol Increases New Bone Formation by Suppression of the Nuclear Factor Kappa B Signaling Pathway

J Craniofac Surg. 2018 Oct;29(7):1983-1990. doi: 10.1097/SCS.0000000000004517.

Abstract

Objective: The objectives of this study were to evaluate the suppression of the nuclear factor kappa B (NF-kB) pathway by 4-hexylresorcinol (4HR), which was activated by tumor necrosis factor-α (TNF-α) in osteoblasts, and new bone formation by 4HR-incorporated porcine bone in an animal model.

Study design: For the confirmation of successful incorporation of 4HR into porcine bone, scanning electron microscopy (SEM) and Fourier transform-infrared (FT-IR) analysis were performed. High performance liquid chromatography was performed for the analysis of the 4HR release profile from porcine bone. MC 3T3-E1 cells were used for the analysis of the NF-kB signaling pathway activation by western blotting and real-time reverse transcriptase polymerase chain reaction. New bone formation and the analysis of marker protein expression were studied in a rat calvarial critical-sized defect model.

Results: Both SEM and FT-IR analysis demonstrated successful incorporation of 4HR into porcine bone. Approximately 30% of 4HR was steadily released from porcine bone for 18 days. 4HR suppressed the NF-kB signaling pathway, which was activated by TNF-α application in MC 3T3-E1 cells. Histological analysis revealed that porcine bone particles with incorporated 4HR showed significantly greater new bone formation than those without 4HR at 4 and 8 weeks after operation (P < 0.05). The expression intensities of alkaline phosphatase, osteoprotegerin, and osteocalcin were also higher in the 4HR-incorporated group.

Conclusion: The application of 4HR suppressed the NF-kB signaling pathway in osteoblasts and 4HR-containing porcine bone particles promoted new bone formation in a rat calvarial defect model.

MeSH terms

  • 3T3 Cells
  • Alkaline Phosphatase / metabolism
  • Animals
  • Blotting, Western
  • Hexylresorcinol / pharmacology*
  • Male
  • Mice
  • Models, Animal
  • NF-kappa B / metabolism*
  • Osteoblasts / drug effects
  • Osteocalcin / metabolism
  • Osteogenesis / drug effects*
  • Osteoprotegerin / metabolism
  • Rats
  • Signal Transduction / drug effects*
  • Spectroscopy, Fourier Transform Infrared
  • Swine
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • NF-kappa B
  • Osteoprotegerin
  • Tnfrsf11b protein, rat
  • Tumor Necrosis Factor-alpha
  • Osteocalcin
  • Alkaline Phosphatase
  • Hexylresorcinol