Recombinant human osteogenic protein-1 (hOP-1) induces new bone formation in vivo with a specific activity comparable with natural bovine osteogenic protein and stimulates osteoblast proliferation and differentiation in vitro.
Recombinant human osteogenic protein-1 (hOP-1) induces new bone formation in vivo with a specific activity comparable with natural bovine osteogenic protein and stimulates osteoblast proliferation and differentiation in vitro.
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发表时间：
1992-10
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通讯作者：
T. K. SampathS;James C. Maliakal;Peter;Hauschkatn;W. Jones;Halina Sasak;Ronald;Tucker;Kerry H. White;J. Coughlin;M. Tucker;H. Roy;Pang;Clare L. C. Corbett;E. Ozkaynak;H. Oppermann;D. Rueger
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作者：
T. K. SampathS;James C. Maliakal;Peter;Hauschkatn;W. Jones;Halina Sasak;Ronald;Tucker;Kerry H. White;J. Coughlin;M. Tucker;H. Roy;Pang;Clare L. C. Corbett;E. Ozkaynak;H. Oppermann;D. Rueger
我们以前报道过，从牛骨基质中纯化的32-36-kDa成骨蛋白由转化生长因子（TGF）-β超家族的两个成员的二聚体组成：人成骨蛋白-1（OP-1）和骨形态发生蛋白-2a（BMP-2a）的牛等价物（BMP-2）。在本研究中，我们在哺乳动物细胞中产生了重组人OP-1（hOP-1），作为一种经加工的成熟二硫键连接的同源二聚体，表观分子量为36，000。在大鼠皮下骨诱导模型中对hOP-1的检查表明，hOP-1能够诱导新骨形成，其比活性与高度纯化的牛成骨蛋白制剂所表现出的比活性相当。hOP-1与大鼠胶原基质制剂组合的半最大骨诱导活性为50-100 ng/25 mg基质，如通过第12天植入物的钙含量测定的。在大鼠成骨细胞富集的骨细胞培养物中，hOP-1对细胞生长和胶原蛋白合成的影响的评价表明，细胞增殖和胶原蛋白合成均以剂量依赖性方式刺激，并且响应于40 ng hOP-1/ml而增加3倍。对成骨细胞表型特征性标志物表达的检测表明，hOP-1特异性刺激碱性磷酸酶的诱导（40 ng hOP-1/ml时增加4倍）、甲状旁腺素介导的细胞内cAMP产生（40 ng hOP-1/ml时增加4倍）和骨钙素合成（25 ng hOP-1/ml时增加5倍）。在β-甘油磷酸盐和L（+）-抗坏血酸盐存在下的成骨细胞的长期（11-17天）培养中，通过每孔的矿物质结节数测量，hOP-1显著增加矿化速率（在20 ng hOP-1/ml时增加20倍）。这些骨细胞培养物中TGF-β 1和hOP-1的直接比较表明，尽管hOP-1和TGF-β 1都促进细胞增殖和胶原合成，但只有hOP-1在特异性刺激成骨细胞表型的标志物方面有效。
We reported previously that a 32-36-kDa osteogenic protein purified from bovine bone matrix is composed of dimers of two members of the transforming growth factor (TGF)-beta superfamily: the bovine equivalent of human osteogenic protein-1 (OP-1) and bone morphogenetic protein-2a, BMP-2a (BMP-2). In the present study, we produced the recombinant human OP-1 (hOP-1) in mammalian cells as a processed mature disulfide-linked homodimer with an apparent molecular weight of 36,000. Examination of hOP-1 in the rat subcutaneous bone induction model demonstrated that hOP-1 was capable of inducing new bone formation with a specific activity comparable with that exhibited by highly purified bovine osteogenic protein preparations. The half-maximal bone-inducing activity of hOP-1 in combination with a rat collagen matrix preparation was 50-100 ng/25 mg of matrix as determined by the calcium content of day 12 implants. Evaluation of hOP-1 effects on cell growth and collagen synthesis in rat osteoblast-enriched bone cell cultures showed that both cell proliferation and collagen synthesis were stimulated in a dose-dependent manner and increased 3-fold in response to 40 ng of hOP-1/ml. Examination of the expression of markers characteristic of the osteoblast phenotype showed that hOP-1 specifically stimulated the induction of alkaline phosphatase (4-fold increase at 40 ng of hOP-1/ml), parathyroid hormone-mediated intracellular cAMP production (4-fold increase at 40 ng of hOP-1/ml), and osteocalcin synthesis (5-fold increase at 25 ng of hOP-1/ml). In long-term (11-17 day) cultures of osteoblasts in the presence of beta-glycerophosphate and L(+)-ascorbate, hOP-1 markedly increased the rate of mineralization as measured by the number of mineral nodules per well (20-fold increase at 20 ng of hOP-1/ml). Direct comparison of TGF-beta 1 and hOP-1 in these bone cell cultures indicated that, although both hOP-1 and TGF-beta 1 promoted cell proliferation and collagen synthesis, only hOP-1 was effective in specifically stimulating markers of the osteoblast phenotype.