Activity of bone morphogenetic protein-7 after treatment at various temperatures: freezing vs. pasteurization vs. allograft

Cryobiology. 2011 Dec;63(3):235-9. doi: 10.1016/j.cryobiol.2011.09.001. Epub 2011 Sep 19.


Insufficient bone union is the occasional complication of biomechanical reconstruction after malignant bone tumor resection using temperature treated tumor bearing bone; freezing, pasteurization, and autoclaving. Since bone morphogenetic protein (BMP) plays an important role in bone formation, we assessed the amount and activity of BMP preserved after several temperature treatments, including -196 and -73°C for 20 min, 60 and 100°C for 30 min, 60°C for 10h following -80°C for 12h as an allograft model, and 4°C as the control. The material extracted from the human femoral bone was treated, and the amount of BMP-7 was analyzed using an enzyme-linked immunosorbent assay. Then, the activity of recombinant human BMP-7 after the treatment was assessed using a bioassay with NIH3T3 cells and immunoblotting analysis to measure the amount of phospho-Smad, one of the signaling substrates that reflect the intracellular reaction of BMPs. Both experiments revealed that BMP-7 was significantly better preserved in the hypothermia groups. The percentages of the amount of BMP-7 in which the control group was set at 100% were 114%, 108%, 70%, 49%, and 53% in the -196, -73, 60, 100°C, and the allograft-model group, respectively. The percentages of the amount of phospho-Smad were 89%, 87%, 24%, 4.9%, and 14% in the -196, -73, 60, 100°C, and the allograft-model group, respectively. These results suggested that freezing possibly preserves osteoinductive ability than hyperthermia treatment.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Biomarkers / analysis
  • Bone Morphogenetic Protein 7 / analysis
  • Bone Morphogenetic Protein 7 / biosynthesis*
  • Bone Morphogenetic Protein 7 / pharmacology
  • Bone Transplantation*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Femur Head / metabolism*
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Freezing
  • Hot Temperature
  • Humans
  • Male
  • Mice
  • NIH 3T3 Cells
  • Pasteurization / methods*
  • Phosphorylation / drug effects
  • Recombinant Proteins / analysis
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / pharmacology
  • Signal Transduction / drug effects
  • Signal Transduction / physiology*
  • Smad1 Protein / metabolism*
  • Transplantation, Autologous
  • Transplantation, Homologous


  • BMP7 protein, human
  • Biomarkers
  • Bone Morphogenetic Protein 7
  • Recombinant Proteins
  • SMAD1 protein, human
  • Smad1 Protein