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The effects of heat on the biological activity of recombinant human bone morphogenetic protein-2
- Source :
- Journal of Bone and Mineral Metabolism. 23:420-425
- Publication Year :
- 2005
- Publisher :
- Springer Science and Business Media LLC, 2005.
-
Abstract
- This study was designed to investigate effects of heat on the bone-inducing activity of recombinant human bone morphogenetic protein (rhBMP)-2. rhBMP-2 samples were heated at 50, 70, 90, or 100 degrees C for 15 min, or 1, 2, 4, or 8 h, or autoclaved at 120 degrees C for 15 min. The bone-inducing activity of the rhBMP-2 before and after heating was assayed in in vivo and in vitro systems. For the in vivo assay, 5 microg rhBMP-2 samples were impregnated into porous collagen disks (6 mm in diameter, 1 mm thickness), freeze dried, and implanted into the back muscles of ddY mice. Three weeks later, the implant was harvested from the host and examined for ectopic new bone tissue by radiography. The new bone mass was quantified by single-energy X-ray absorptiometry. The in vitro activity of the rhBMP-2 was assayed by adding the BMP sample at a concentration of 100 ng/ml to cultures of MC3T3-E1 cells. After 48 h, the alkaline phosphatase activity was measured. After heating at 50 degrees or 70 degrees C, no significant reduction in bone-inducing activity was noted in either in vivo or in vitro assay systems unless the protein was exposed to sustained heat at 70 degrees C for 8 h, based on in vitro assay data. However, heating above 90 degrees C and for longer periods led to a decrease in the biological activity of the rhBMP-2 in a time- and temperature-dependent manner. rhBMP-2 was rendered inactive when exposed to temperatures at or in excess of 120 degrees C.
- Subjects :
- Hot Temperature
Endocrinology, Diabetes and Metabolism
Bone Morphogenetic Protein 2
Bone morphogenetic protein
Bone tissue
Bone morphogenetic protein 2
Cell Line
Mice
Calcification, Physiologic
Endocrinology
Transforming Growth Factor beta
In vivo
medicine
Animals
Humans
Orthopedics and Sports Medicine
Chemistry
Biological activity
General Medicine
Alkaline Phosphatase
Molecular biology
Recombinant Proteins
In vitro
medicine.anatomical_structure
Biochemistry
Cell culture
Enzyme Induction
Bone Morphogenetic Proteins
Alkaline phosphatase
Electrophoresis, Polyacrylamide Gel
Subjects
Details
- ISSN :
- 14355604 and 09148779
- Volume :
- 23
- Database :
- OpenAIRE
- Journal :
- Journal of Bone and Mineral Metabolism
- Accession number :
- edsair.doi.dedup.....96198bd8ff29c59bebcf9d9a7b95106e