Collagen‐targeted BMP3 fusion proteins arrayed on collagen matrices or porous ceramics impregnated with Type I collagen enhance osteogenesis in a rat cranial defect model

Journal of Orthopaedic Research - Tập 20 - Trang 747-755 - 2002
Bo Han1, Natalya Perelman1, Baowei Tang1, Fredrick Hall1, Edwin C Shors1,2, Marcel E Nimni1
1Surgical Research Laboratories, Departments of Surgery, Orthopaedics and Biochemistry, University of Southern California, Keck School of Medicine, HMR-810, Los Angeles, CA 90033, USA
2Interpore-Cross International, Irvine, CA 92718, USA

Tóm tắt

Abstract

Bone morphogenetic protein 3 (BMP3) is a potent osteoinductive growth factor belonging to the TGF‐β superfamily. In this study, we engineered a recombinant BMP3 protein to include an auxiliary collagen‐targeting domain derived from von Willebrand coagulation factor (vWF). The collagen‐targeted BMP3 fusion protein (rhBMP3‐C) was expressed in E. coli, purified from bacterial inclusion bodies, renatured under controlled redox conditions, and assayed for biological activity in vitro and in vivo. The renatured rhBMP3‐C fusion protein bound tightly to collagen matrices and inhibited DNA synthesis in normal rat calvaria cells and in two out of three human osteosarcoma cell lines tested. Alkaline phosphatase activity was increased in rat calvarial cells and was decreased in osteosarcoma cells in vitro in a dose‐dependent manner. Collagen sponges impregnated with rhBMP3‐C and implanted subcutaneously in Fischer‐344 rats induced dose‐dependent dystrophic calcification of the collagen matrix, with no evidence of ectopic bone formation. However, local injection of rhBMP3‐C infused in a collagen suspension induced new bone formation on the periosteal surface of rat calvaria. Finally, in a rat cranial defect model, surgical implantation of rhBMP3‐C arrayed on either collagen sponges or on porous ceramics coated with Type I collagen exhibited marked osteoinductive properties. Taken together, these results demonstrate the feasibility of engineering and manufacturing targeted‐BMPs which exhibit an integral gain‐of‐function that may be exploited to therapeutic advantage in (i) the enhancement of effective local concentrations, (ii) the prevention of systemic biodistribution and side effects, and (iii) the design of improved osteoinductive matrices. © 2002 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.


Tài liệu tham khảo

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Journal of Orthopaedic Research

Tập 20

747-755

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