title: Activity of bone morphogenetic protein-7 after treatment at various temperatures: freezing vs. pasteurization vs. allograft.
abstract: 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.
authors.affiliation: Department of Orthopaedic Surgery, Graduate School of Medical Science, Kanazawa University, Kanazawa, Japan.
authors.forename: Munetomo
authors.initials: M
authors.lastname: Takata
authors.forename: Naotoshi
authors.initials: N
authors.lastname: Sugimoto
authors.forename: Norio
authors.initials: N
authors.lastname: Yamamoto
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Shirai
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Hayashi
authors.forename: Hideji
authors.initials: H
authors.lastname: Nishida
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Tanzawa
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kimura
authors.forename: Shinji
authors.initials: S
authors.lastname: Miwa
authors.forename: Akihiko
authors.initials: A
authors.lastname: Takeuchi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tsuchiya
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D055419:Bone Morphogenetic Protein 7 / Q000032:analysis / Q000096:biosynthesis* / Q000494:pharmacology; D016025:Bone Transplantation; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005270:Femur Head / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005615:Freezing; D006358:Hot Temperature; D006801:Humans; D008297:Male; D051379:Mice; D041681:NIH 3T3 Cells; D061352:Pasteurization / Q000379:methods*; D010766:Phosphorylation / Q000187:drug effects; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D051898:Smad1 Protein / Q000378:metabolism*; D014182:Transplantation, Autologous; D014184:Transplantation, Homologous
chemical_list: C525819:BMP7 protein, human; D015415:Biomarkers; D055419:Bone Morphogenetic Protein 7; D011994:Recombinant Proteins; C497399:SMAD1 protein, human; D051898:Smad1 Protein