Reference - Detail
| RRC ID | 19321 |
|---|---|
| Author | Yano K, Hoshino M, Ohta Y, Manaka T, Naka Y, Imai Y, Sebald W, Takaoka K. |
| Title | Osteoinductive capacity and heat stability of recombinant human bone morphogenetic protein-2 produced by Escherichia coli and dimerized by biochemical processing. |
| Journal | J Bone Miner Metab |
| Abstract |
One problem associated with clinical application of CHO-derived recombinant human bone morphogenetic protein (C-BMP-2) is its high cost due to the need for use of high doses. To solve this problem, Escherichia coli-derived BMP-2 (E-BMP-2) has been examined using the technique of molecular unfolding and refolding. However, it is unclear whether the characteristics of E-BMP-2 are appropriate for clinical application. In this study, we examined the biological activity of E-BMP-2 and its heat tolerance in in vitro and in vivo systems. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) confirmed the high purity of E-BMP-2. E-BMP-2-induced alkaline phosphatase expression in osteoprogenitor cells (C2C12, ST2, and primary murine calvarial osteoblast cells) was dose-dependent, and consistently elicited ectopic new ossicles of significant size in mice, also in dose-dependent fashion. In addition, E-BMP-2 induced phosphorylation of Smad1/5/8 and mRNA expression of osteoblastic differentiation markers to the same extent as C-BMP-2. On the other hand, when E-BMP-2 was exposed to increasing heat over time, its bone-inducing capacity was maintained until reaching 70 degrees C for 2 h or 90 degrees C for 15 min. Thus, E-BMP-2 will exhibit a decrease in activity with the sterilization procedures required prior to use in surgery. These findings indicate that the biological capacity and heat stability of E-BMP-2 are almost equivalent to those of currently available C-BMP-2, and suggest that E-BMP-2 might, thus, solve current problems of cost impeding routine clinical use of rhBMP-2. |
| Volume | 27(3) |
| Pages | 355-63 |
| Published | 2009-1-1 |
| DOI | 10.1007/s00774-009-0040-3 |
| PMID | 19229473 |
| MeSH | Alkaline Phosphatase / biosynthesis Animals Biochemical Phenomena / drug effects* Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins / biosynthesis Bone Morphogenetic Proteins / pharmacology* Bone and Bones / diagnostic imaging Bone and Bones / drug effects Cell Differentiation / drug effects Cells, Cultured Electrophoresis, Polyacrylamide Gel Enzyme Induction / drug effects Escherichia coli / metabolism* Hot Temperature* Humans Mice Mice, Inbred ICR Osteoblasts / cytology Osteoblasts / drug effects Osteoblasts / enzymology Osteogenesis / drug effects* Protein Multimerization / drug effects* Protein Stability / drug effects* Radiography Recombinant Proteins / biosynthesis Recombinant Proteins / pharmacology* Smad Proteins / metabolism Transforming Growth Factor beta / biosynthesis Transforming Growth Factor beta / pharmacology* |
| IF | 2.297 |
| Times Cited | 47 |
| WOS Category | MEDICINE, RESEARCH & EXPERIMENTAL ENDOCRINOLOGY & METABOLISM |
| Altmetric score |
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 2 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | ST2(RCB0224) C2C12(RCB0987) |