Application of newly-developed gene transfer to regeneration of bone and periodontal tissue

新开发的基因转移在骨和牙周组织再生中的应用

• 批准号: 16390579
• 负责人: KASUGAI Shohei
• 金额: $ 9.09万
• 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 Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390579/
• 关键词: Gene Transfer; Plasmid DNA; BMP; Bone; Periodontal tissue; Tissue Regeneration; Collagen; Ligament; プラスミドDNA; 再生医療

As regenerative therapies for bone and periodontal tissues several methods have been developed and clinically applied ; however, a method, which is simple, safe and economical for public society, is required. Although gene transfer with plasmid DNA is safe and clinically applied, it is a problem that efficiency of this gene transfer is extremely low. In the present study, we obtained the following results.1. When plasmid DNA was combined with small Ca-phosphate particles and transplanted to the tissue defect, the efficiency of gene transfer increased. Small Ca-phosphate particles bound plasmid DNA and kept it locally. Furthermore, the binding to the particles increased the resistance to DNase. These phenomena would contribute to the increase of gene transfer.2. Utilizing this method, we applied plasmid DNA encoding human BMP2 to 5 mm segmental bone defects of rat tibias. Six weeks the defects completely healed and the mechanical strength of the bone was comparable to the normal tibia at 8 weeks. Although knot-like bone appeared at the applied site, it was gradually absorbed and the bone shape became normal.3. Similarly, plasmid DNA encoding human BMP12 was applied to the bone defects of rat femurs. Ligament-like tissue abundant in collagen fibers was formed from the previous defect to the surrounding tissue whereas the defect was filled with bone. This method would be promising for ligament reconstruction.4. Periosteum cells were cultured and these cells were transferred with BMP2 and/or VEGF gene, which were then transplanted to nude mice. When both genes were transferred, the largest amount of bone was formed.

作为骨和牙周组织的再生疗法，已经开发并临床应用了几种方法;然而，需要一种对公众社会来说简单、安全和经济的方法。尽管用质粒DNA进行基因转移是安全的并且临床上可应用，但问题是这种基因转移的效率极低。在本研究中，我们获得了以下结果.当质粒DNA与磷酸钙小颗粒结合并移植到组织缺损处时，基因转移的效率增加。小的磷酸钙颗粒结合质粒DNA并将其保持在局部。此外，与颗粒的结合增加了对DNA酶的抗性。这些现象将有助于基因转移的增加.利用这种方法，我们应用质粒DNA编码的人骨形成蛋白2到5毫米节段性骨缺损的大鼠胫骨。6周时缺损完全愈合，8周时骨的机械强度与正常胫骨相当。虽然在应用部位出现结节状骨，但逐渐被吸收，骨形状变得正常。类似地，将编码人BMP 12的质粒DNA应用于大鼠股骨的骨缺损。胶原纤维丰富的韧带样组织从先前的缺损到周围组织形成，而缺损被骨填充。该方法在韧带重建中有应用前景.培养骨膜细胞，将BMP 2和/或VEGF基因转入这些细胞，然后移植到裸小鼠体内。当两种基因都被转移时，最大量的骨骼形成。

项目成果

Effectiveness of Extracted-teeth as Bone Substitute : Application to Parietal Bone Defects in Rabbits.

拔牙作为骨替代品的有效性：应用于兔顶骨缺损。

• 期刊: Journal of Oral Tissue Engineering (In Press)
• 作者: K.Tint;H.Kondo;S.Kuroda;T.Fujimori;K.Aoki;K.Ohya;S.Kasugai
• 通讯作者: S.Kasugai

骨造成法の現状と近未来.

骨移植方法的现状和近期前景。

• 发表时间: 2005
• 期刊: 日本補綴学会雑誌 49(5)
• 作者: Katakura O;Morimoto N;Iwasaki Y;Akiyoshi K;Kasugai S;春日井昇平
• 通讯作者: 春日井昇平

Bone regeneration by modified gene-activated matrix: Effectiveness in segmental tibial defects in rats

• DOI: 10.1089/ten.2006.12.489
• 发表时间: 2006-03-01
• 期刊: TISSUE ENGINEERING
• 作者: Endo, M;Kuroda, S;Kasugai, S
• 通讯作者: Kasugai, S

現代歯科薬理学

现代牙科药理学

• 发表时间: 2005
• 作者: Nakamura S;Nakamura M;Kobayashi T;Sekijima Y;Kasugai S;Yamashita K;春日井昇平(共著)
• 通讯作者: 春日井昇平(共著)

Bone increase in rat tibiae by local administration of amino-terminally truncated rhFGF-4(73-206).

通过局部施用氨基末端截短的 rhFGF-4(73-206) 增加大鼠胫骨的骨量。

• 发表时间: 2007
• 期刊: Tissue Engineering 13・2
• 作者: Shinji Kuroda;Hisatomo Kondo;Keiichi Ohya;Shohei Kasugai
• 通讯作者: Shohei Kasugai