Inductive properties of recombinant human BMP-2 produced in a bacterial expression system

• Published in: International journal of oral and maxillofacial surgery 1998-08, Vol.27 (4), p.305-309
• Main Authors: Kübler, N.R., Reuther, J.F., Faller, G., Kirchner, T., Ruppert, R., Sebald, W.
• Format: Article
• Language: English
• Subjects: Abdominal Muscles - drug effects; Abdominal Muscles - metabolism; Animals; Biological and medical sciences; Biotechnology; BMP-2; bone formation; Bone Morphogenetic Protein 2; bone morphogenetic proteins; Bone Morphogenetic Proteins - administration & dosage; Bone Morphogenetic Proteins - genetics; Bone Morphogenetic Proteins - pharmacology; Cartilage - drug effects; Cartilage - metabolism; Cell Differentiation; Chondrocytes - drug effects; Chondrocytes - metabolism; Chondrocytes - pathology; Collagen; Culture Techniques; Dentistry; Dose-Response Relationship, Drug; Drug Carriers; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Bacterial; Health. Pharmaceutical industry; Hormones. Growth factors; Humans; Industrial applications and implications. Economical aspects; Mesoderm - drug effects; Mesoderm - metabolism; Mesoderm - pathology; Mice; Mice, Inbred ICR; Muscle, Skeletal - drug effects; Muscle, Skeletal - metabolism; Ossification, Heterotopic - etiology; Ossification, Heterotopic - metabolism; Ossification, Heterotopic - pathology; Osteogenesis - drug effects; osteoinduction; Production of active biomolecules; Proteins - metabolism; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Transforming Growth Factor beta - administration & dosage; Transforming Growth Factor beta - genetics; Transforming Growth Factor beta - pharmacology
• ISSN: 0901-5027; 1399-0020
• DOI: 10.1016/S0901-5027(05)80621-6

Recombinant human BMP-2, produced in E. coli, refolded and concentrated to a purity of more than 98%, has been demonstrated to be biologically active. In vitro, amounts of 0.4 μg BMP-2 or more induced new cartilage formation in 27 out of 47 samples of a neonatal muscle tissue assay, with chondroneogenesis occurring 14 days after a four-hour contact between BMP-2 and the muscle tissue. In vivo, BMP-2 was implanted in the thigh muscle of ICR mice for a period of three weeks. Amounts of 4 μg BMP-2 and more showed heterotopic bone formation in 15 out of 17 samples. When BMP-2 was combined with a collagen carrier, amounts of 0.4 μg protein or more induced heterotopic bone formation in 30 out of 33 samples four weeks after the implantation in the abdominal wall of Sprague-Dawley rats. The results show that the E. coli-derived BMP-2 was active in different assay systems in concentrations equal to those required with mammalian cell-expressed BMP-2. It could also be demonstrated that a single morphogen (BMP-2) is enough to initiate the differentiation process associated with bone induction. The presented bacterial expression system also offers the opportunity to produce large quantities of recombinant BMP-2 for clinical applications.