Immobilized recombinant human bone morphogenetic protein-2 enhances the phosphorylation of receptor-activated Smads.
Bone morphogenetic protein (BMP)-2 plays an important role in bone growth and regeneration; however, BMP-2 is easily lost by diffusion through body fluid and has some inhibitory pathways. To address this problem, we previously immobilized recombinant human BMP-2 (rhBMP-2) on succinylated type I atelocollagen. Here, we examined the effect of immobilized rhBMP-2 in vitro and vivo. In ST2, MC3T3-E1, and C2C12 cells, alkaline phosphatase activity, which is a marker of osteoblast differentiation, was enhanced more by immobilized than nonimmobilized rhBMP-2. In addition, the phosphorylation of receptor-activated Smads, part of the signaling pathway activated by BMP-2, was prolonged by immobilized rhBMP-2 in these cells. Furthermore, implantation of immobilized rhBMP-2 into the backs of rats promoted the formation of mature bone-like structure. These results demonstrate that immobilized rhBMP-2 has higher bioactivity than nonimmobilized rhBMP-2, and, therefore, immobilization of rhBMP-2 can prolong BMP signaling.
Yamachika
Eiki
E
Department of Oral and Maxillofacial Reconstructive Surgery, Graduate school of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikata-Cho, Okayama 700-8525, Japan. eikiyama@md.okayama-u.ac.jp
Tsujigiwa
Hidetsugu
H
Shirasu
Nobuaki
N
Ueno
Takaaki
T
Sakata
Yoshirou
Y
Fukunaga
Joji
J
Mizukawa
Nobuyoshi
N
Yamada
Masao
M
Sugahara
Toshio
T
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D024042:Collagen Type I / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D051379:Mice; D015675:Osteocalcin / Q000235:genetics; D053495:Osteopontin / Q000235:genetics; D010766:Phosphorylation / Q000187:drug effects; D019919:Prosthesis Implantation; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology*; D051785:Smad Proteins / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000737:chemistry / Q000494:pharmacology*
D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D024042:Collagen Type I; D012333:RNA, Messenger; D011994:Recombinant Proteins; D051785:Smad Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; D015675:Osteocalcin; D053495:Osteopontin; D000469:Alkaline Phosphatase