Development of Bioactive Pulp Capping Agent

生物活性盖髓剂的研制

• 批准号: 04557082
• 负责人: NAGASAWA Hisashi
• 金额: $ 9.98万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04557082/
• 关键词: BONE Morphogenetic Protein; Transforming growth factor; pulp cells; Odontoblasts; Reparative Dentin; Cell culture; mRNA; Amputation; 歯髄細胞培養; ノーザンブロット; コラーゲンI型; コラーゲンIII型; オステオカルシン; アルカリフォスファターゼ; 象牙芽細胞誘導因子

The expression of developmental stage specific genes during pulp cell differentiation into preodontoblasts was examined in bovine adult pulp cell culture. When proliferation was down-regulated after 14 days, expression of fibronectin, type I and type III collagen mRNA was increased. Expression of alkaline phosphatase was gradually increased, and mRNA for osteocalcin, a marker of preodontoblast, appeared just before onset of mineralization. Expression of TGF-beta1 mRNA disappeared on day 21 when expression of alkaline phosphatase mRNA was increased. BMP-4 mRNA was expressed on day 14 when the expression of the extracellular matrix proteins was increased. BMP-2 mRNA was expressed on day 28 when osteocalcin appeared. Recombinant TGF-beta1 inhibited alkaline phosphatase activity, while BMP-2 and BMP-4 stimulated it. BMP-4 increased expression of alpha1(I) collagen mRNA, and BMP-2 increased osteocalcin synthesis. These results demonstrate the regulatory role of these TGF-beta superfamily members on the gene expression of extracellular matrix proteins and the differentiation of pulp cells into preodontoblasts.BMP-2 and -4 and TGF-beta1 were applied in the cavity on amputated pulp to examine their effects on pulp regeneration and reparative dentin formation. When BMP-2 and -4 were combined with type I collagen as a carrier, osteodentin was formed in the cavity 2 months postoperatively. While TGF-beta1 had no effect. In BMP-2 and -4 with inactivated dentin matrix applied teeth, tubular dentin was formed in the lower part of the cavity, although osteodentin was formed in the upper part, suggesting that some matrix component might be essential for differentiation into odontoblasts in addition to BMPs.We are going to develop an ideal carrier for the use of BMPs as a biologically active pulp capping agent.

在牛成年牙髓细胞培养中，研究了牙髓细胞向前成牙本质细胞分化过程中发育阶段特异性基因的表达。14天后细胞增殖下调，纤维连接蛋白、I型和III型胶原mRNA表达增加。碱性磷酸酶的表达逐渐增加，骨钙素，前成牙本质细胞的标志物，mRNA出现在矿化开始之前。TGF-β 1 mRNA的表达在第21天消失，而碱性磷酸酶mRNA的表达增加。BMP-4 mRNA在第14天表达，此时细胞外基质蛋白的表达增加。BMP-2 mRNA在第28天骨钙素出现时表达。重组TGF-β 1抑制碱性磷酸酶活性，而BMP-2和BMP-4刺激碱性磷酸酶活性，BMP-4增加α 1（I）胶原mRNA的表达，BMP-2增加骨钙素的合成。BMP-2、BMP-4和TGF-β 1对牙髓再生和修复性牙本质形成的影响。当BMP-2和BMP-4与I型胶原复合作为载体时，术后2个月骨牙本质在腔中形成。而TGF-β 1则没有影响。在BMP-2和BMP-4与失活的牙本质基质的作用下，牙洞下部形成管状牙本质，而牙洞上部形成骨牙本质，提示除了BMP外，某些基质成分可能是成牙本质细胞分化所必需的。

项目成果

Nakashima,M.: "Reparative dentin induction on canine amputated pulp by recombinant human bone morphogenetic protein(BMP)-2 and 4." J.Dent Res. (in press).

Nakashima,M.：“通过重组人骨形态发生蛋白 (BMP)-2 和 4 对犬截肢牙髓进行修复性牙本质诱导。”

Nakashima M.,Nagasawa,H.,Yamada Y.,Reddi A.H.: "Regulatory role of Transforminng growth factor-β,Bone morphogenetic protein-2 and protein-4 on gene expression of extracellular matrix proteins and differentiation of dental pulp cells." Developmental Biolog

Nakashima M.、Nagasawa, H.、Yamada Y.、Reddi A.H.：“转化生长因子-β、骨形态发生蛋白-2 和蛋白-4 对细胞外基质蛋白基因表达和牙髓细胞分化的调节作用。”发育生物学

Nakashima M., Nagasawa H., Yamada T., Reddi A.H.: "Regulatory role of Transforming growth factor-beta, Bone morphogenetic protein-2 and protein-4 on gene expression of extracellular matrix proteins and differentiation of dental pulp cells." Dev Biol. 162.

Nakashima M.、Nagasawa H.、Yamada T.、Reddi A.H.：“转化生长因子-β、骨形态发生蛋白-2 和蛋白-4 对细胞外基质蛋白基因表达和牙髓细胞分化的调节作用。”

Nakashima,M.: "Induction of dentine in amputated pulp of the dog by recombinant human bone morphogenetic protein(BMP)-2 and -4 with collagen matrix." Archs oral Biol. (in press).

Nakashima,M.：“通过重组人骨形态发生蛋白 (BMP)-2 和 -4 与胶原基质诱导狗断牙髓中的牙本质。”