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

作者：Takata Munetomo; Sugimoto Naotoshi; Yamamoto Norio; Shirai Toshiharu; Hayashi Katsuhiro; Nishida Hideji; Tanzawa Yoshikazu; Kimura Hiroaki; Miwa Shinji; Takeuchi Akihiko; Tsuchiya Hiroyuki^*

来源：Cryobiology, 2011, 63(3): 235-239.

DOI：10.1016/j.cryobiol.2011.09.001

• Bone tumor
• Freezing
• Bone morphogenetic protein
• Protein stability
• Allograft
• Liquid nitrogen
• Cryotherapy
• Osteoinductive ability

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 degrees C for 20 min, 60 and 100 degrees C for 30 min, 60 degrees C for 10 h following -80 degrees C for 12 h as an allograft model, and 4 degrees 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 degrees 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 degrees C, and the allograft-model group, respectively. These results suggested that freezing possibly preserves osteoinductive ability than hyperthermia treatment.

• 出版日期2011-12