Research and Development of New Bone Substitute Using BMP with Atelocollagen

使用 BMP 和去端肽胶原的新型骨替代品的研究与开发

• 批准号: 63870078
• 负责人: ENOMOTO Shoji
• 金额: $ 4.03万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63870078/
• 关键词: Bone Substitute; BMP; Hydroxyapatite; Atelocollagen; Reconstruction of Mandible; Bone-Induction; BMP; アテロコラーゲン; 複合材

1. Purification of Bone Morphogenetic Protein (BMP): 1) A crude BMP, which was extracted from decalcified bovine bone, was separated with gel filtration, cation exchange chromatography, heparin affinity chromatography, and finally separated with reverse-phase HPLC. SDS-PAGE erectrophoretic examination of the final BMP-fraction showed two smear bands of 16kDa and 30-40kDa under non- reducing conditions, 16kDa and 18kDa tinder reducing conditions. Amino acid sequence analysis revealed the similarity of the protein with a molecular weight of 16kDa to BMP-3 which was reported by J. Wazney et al. in 1988.2) Purification of BMP extracted from human teeth was also performed in the same way as bovine bone. Separation with a series of chromatographies glowed the similarity of elution patterns to that of bovine bone. However, the final purification is under investigation.2. Examination of BMP-composite (AC-BMP, HA-AC-BMP) by implantation into bone- defects: 1)Aterocoliagen-BMP composite (AC-BMP) … More and liydroxyapatite-atelocollagen -BMP composite (HA-AC-BMP) (both were lyophilized and compressed), HA/AC/BMP mixture (gel) were implanted intoskull defects of rats. Both AC-GMP composite and HA-AC-BMP composite consistently induced the most remarkable new-bone formation from early stage. The repair of defects which had been implanted with AC-BMP composite and HA-AC-BMP composite were the most rapid and complete. 2) HA-AC-BMP composite was also implanted into mandible of monkey. Three months after implantation, the defect was completely regenerated by newly formed bone in the implantation of RA-AC-BMP composite, while small amount of new bone was formed only at the rim of the host bed in the implantation of HA-AC composite as control.These results indicate that AC-BMP composite and HA-AC-BMP composite have excellent bone-inducing and bone-regenerating potency. Results from the Primates suggest the possibility of these BMP-composite (AC-BMP, HA-AC-BMP) inclinical use as bone-inductive biomaterials in place of fresh autogenous bone. Less

1.骨形态发生蛋白（BMP）的纯化：1）从脱钙牛骨中提取BMP粗品，经凝胶过滤、阳离子交换层析、肝素亲和层析、反相高效液相色谱分离。最终BMP-级分的SDS-PAGE电泳检测在非还原条件下显示16 kDa和30- 40 kDa的两条涂抹带，在还原条件下显示16 kDa和18 kDa。氨基酸序列分析表明，该蛋白质分子量为16 kDa，与J. Wazney等1988年报道的BMP-3相似。2）从人牙中提取的BMP的纯化方法与牛骨相同。用一系列色谱分离，使洗脱模式与牛骨相似。然而，最终的纯化正在研究中。BMP-复合物（AC-BMP、HA-AC-BMP）植入骨缺损的检查：1）动脉胶原-BMP复合物（AC-BMP） ...更多信息 将HA/AC/BMP混合物（凝胶）和HA/AC/BMP复合物（HA-AC-BMP）植入大鼠颅骨缺损区。AC-GMP复合材料和HA-AC-BMP复合材料在早期诱导新骨形成最为显著。AC-BMP复合材料和HA-AC-BMP复合材料修复骨缺损最快、最完全。2)将HA-AC-BMP复合材料植入猴下颌骨内。3个月后，RA-AC-BMP复合材料可使骨缺损完全再生，而HA-AC复合材料仅在宿主骨床边缘有少量新骨形成，表明AC-BMP复合材料和HA-AC-BMP复合材料具有良好的骨诱导和骨再生能力。实验结果表明，这些BMP-复合材料（AC-BMP、HA-AC-BMP）有可能作为替代新鲜自体骨的骨诱导生物材料。少

项目成果

Masatoshi Watanabe: Handbook of Bioactive Ceramics, vol II, Editor: T.Yamamuro, J.Wilson, L.L.Hench chapter 22: Implantation of Hydroxyapatite Granule with Atelocollagen and Bone Inductive Protein in rat Skull Defect. CRC Press, Inc., U.S.A.,

Masatoshi Watanabe：《生物活性陶瓷手册》，第二卷，编辑：T.Yamamuro、J.Wilson、L.L.Hench 第 22 章：在大鼠颅骨缺损中植入含去端胶原和骨诱导蛋白的羟基磷灰石颗粒。

Masatoshi Watanabe Editor:T.Yamamuro,L.L.Hench,J.Wilson: "Handbook of Bioactive Ceramics,vol II,chapter 22(in press).Implantation of Hydroxyapatite Granule with Atelocollagen and Bone Inductive Pritein in rat Skull Defect." CRC Press,Inc.,U.S.A., (1990)

Masatoshi Watanabe 编辑：T.Yamamuro、L.L.Hench、J.Wilson：“生物活性陶瓷手册，第二卷，第 22 章（待出版）。含去端肽胶原和骨诱导蛋白的羟基磷灰石颗粒在大鼠颅骨缺损中的植入。”

渡辺正敏: "ハイドロキシアパタイト・アテロコラ-ゲン・骨形成因子複合材のラット頭蓋骨欠損部移植による組織学的検討" 日本口腔外科学会雑誌.

Masatoshi Watanabe：“羟基磷灰石、去端肽胶原和骨形态发生因子复合物移植到大鼠颅骨缺损中的组织学研究”日本口腔颌面外科学会杂志。

渡辺正敏: "ハイドロキシアパタイト・アテロコラ-ゲン・骨形成因子複合材のラット頭蓋骨欠損部移植による組織学的検討" 日本口腔外科学会雑誌(投稿中).

Masatoshi Watanabe：“羟基磷灰石、去端肽胶原和骨形态发生因子复合物移植到大鼠颅骨缺损中的组织学研究”日本口腔颌面外科学会杂志（目前正在提交）。

Masatoshi Watanabe Editor:T.Yamamuro L.L.Hench J.Wilson: "Handbook of Bioactive Ceramics,vol.II,chapter 22(in press).Implantation of Hydroxyapatite Granule with Atelocollagen and Bone Inductive Pritein in rat Skull Defect." CRC Press,Inc.,U.S.A., (1990)

Masatoshi Watanabe 编辑：T.Yamamuro L.L.Hench J.Wilson：“生物活性陶瓷手册，第 II 卷，第 22 章（待出版）。含去端肽胶原和骨诱导蛋白的羟基磷灰石颗粒在大鼠颅骨缺损中的植入。”