Experimental study of osteoinduction with gene transfer of bone morphogenetic protein using adenovirus vector.

使用腺病毒载体进行骨形态发生蛋白基因转移的骨诱导实验研究。

• 批准号: 15591910
• 负责人: KUSUMOTO Kenji
• 金额: $ 2.24万
• 依托单位: Kansai Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591910/
• 关键词: bone morphogenetic protein; osteoinduction; adenovirus vector; bone formation; bone marrow; gene transfer; tissue transfer; regenerative medicine; アデノウイルスベクター; 骨再建; 再生医学; サイトカイン

Bone morphogenetic protein (BMP) has a unique ability, osteoinduction. BMP induces bone in the soft tissue. It is expected that BMP is applied in bone reconstruction. In this study, BMP is implanted in soft tissue, and adenovirus vector with BMP-2 gene (AVV-rhBMP-2) is transfected in the muscle. The induced bone is investigated.1)rhBMP-2 (2,10,50ug) and atelopeptide tipe I collagen were mixed and freezed-dried (rhBMP-2-CL).2)rhBMP-2 gene was associated into cosmid vector. On the other hand, adenovirus deficit Δ1Δ3 and cosmid vector were co-transfected to HEK293 cells. In the cells, AVV-rhBMP-2 was multiplicated and AVV-rhBMP-2 was purified.3)muscle flaps were operated on the back of rats. There rhBMP-2 and AVV-rhBMP-2 were implanted. Three weeks later the implanted areas soft X-rayed and the implants were excised and examined histologically and biochemically.4)The implanted areas were excised and HE-stained to investigate the tissues histologically.In both, induced bone were investigated. rhBMP-2-CL induced massive bone including chondrocytes in 10days, bony mass with bone marrow in 21 days. AVV-rhBMP-2 induced scattered bony particles. Each was bone mass. Comparing controls, the amount of induced bone was dose-dependent in both groups.

骨形态发生蛋白（BMP）具有独特的骨诱导能力。BMP在软组织中诱导骨。BMP有望应用于骨重建。本研究将BMP-2植入软组织，并将携带BMP-2基因的腺病毒载体（AVV-rhBMP-2）转染肌肉。1）将rhBMP-2（2、10、50 μ g）与端肽Ⅰ型胶原混合冻干（rhBMP-2-CL）; 2）将rhBMP-2基因连接到粘粒载体中。另一方面，将腺病毒缺陷型Δ1Δ3和粘粒载体共转染到HEK 293细胞中。在细胞中扩增AVV-rhBMP-2，并纯化AVV-rhBMP-2。3）在大鼠背部肌瓣移植。植入rhBMP-2和AVV-rhBMP-2。三周后，对植入区域进行软X光检查，并切除植入物并进行组织学和生物化学检查。4）切除植入区域并进行HE染色以进行组织学研究。在两种情况下，均对诱导骨进行了研究。rhBMP-2-CL在10天内诱导成大量骨，包括软骨细胞，在21天内诱导成含有骨髓的骨块。AVV-rhBMP-2诱导骨颗粒分散。每一个都是骨量。与对照组相比，两组中诱导骨的量呈剂量依赖性。

项目成果

Preclinical study of recombinant human bone morphogenetic protein-2 : Application of hyperbaric oxygenation during bone formation under unfavorable condition.

重组人骨形态发生蛋白2的临床前研究：高压氧在不利条件下骨形成过程中的应用。

• 发表时间: 2003
• 期刊: Int J Oral Maxillo fac Surg 32
• 作者: Okubo Y;Bessho K;Fujimura K;Kusumoto K;Suzuki T;Segami N;Ogawa Y;Iizuka T.
• 通讯作者: Iizuka T.

外鼻変形に対する脂肪注入・移植

脂肪注射/移植治疗外鼻畸形

• 发表时间: 2005
• 期刊: 洛和会病院医学雑誌 16
• 作者: 田辺敦子;三宅省吾;中島大毅;楠本健司;小川 豊
• 通讯作者: 小川 豊

Kusumoto K, Bessho K: "Bone morphogenetic protein and vascular endothelial growth factor in bone formation"Research Signpost, Trivandrum. 14 (2003)

Kusumoto K、Bessho K：“骨形态发生蛋白和血管内皮生长因子在骨形成中的作用”研究路标，特里凡得琅。

骨再建材としての体内埋入異物の問題点と展望

体内异物植入作为骨重建材料存在的问题及前景

• 发表时间: 2003
• 期刊: 日本美容外科学会会報 25
• 作者: 楠本健司;小川 豊
• 通讯作者: 小川 豊

ヒト線維芽細胞培養における各種血清の比較およびTGF-β含有量の検討

人成纤维细胞培养物中各种血清比较及TGF-β含量检测

• 发表时间: 2005
• 期刊: 熱傷 31(1)
• 作者: 佐々木富美子;楠本健司;小川 豊
• 通讯作者: 小川 豊