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Development of a novel therapeutic agent of bone using synthetic peptide of ameloblastin

使用成釉细胞合成肽开发新型骨治疗剂

基本信息

批准号：
17390486
负责人：
TAKATA Takashi
金额：
$ 10.81万
依托单位：
Hiroshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

项目摘要

To determine whether synthetic peptide of the N-terminal of ameloblastin as a novel regenerative agent for bore defect, we achieved in vitro and in vivo analyses and obtained the following results.1. To determine the ameloblastin fragment with the most active biological effects on osteoblastic cells, we examined activities of alkaline phosphate and calcification of osteoblastic cell line, MC3T3-E1, after treatment of various lengths of N-terminal area of ameloblastin. It was show that the highest inductive effects were seem when the cells treated with 16 amino acids (VPFFPQQSGTPGMASL) at N-terminal of ameloblastin (16N).2. To examine the signaling pathway of 16N in osreoblasts, expression of MAPK in MC3T3-E1 cells after the treatment of 16N. Phosphotylation of p38 was seen in the cells.3. To recognize the receptor of 16N, we transfected cDNA encoding 16N into MC3T3-E1cells. A vector with the sequence of secretion could induce high alkaline phosphatase activity in MC3T3-E1 cells, while a vector without the sequence of secretion didn't induced. It was, therefore speculated that the receptor of 16N may present on the surface of MC3T3-E1 cells.4. Although identification of the membrane located receptor was not accomplished, CD63was detected on the cell membrane of MC3T3-E1 cells and suspected as a potential receptor of 16N.5. In vivo studies with different bone defect models showed regeneration promoting effects on 16N. These findings indicate that 16N can be used for novel agent promoter of bone healing

为确定成釉蛋白N端合成肽是否能作为一种新型的孔洞再生剂，我们进行了体外和体内的分析，并获得了以下结果。为了确定对成骨细胞具有最活跃生物学作用的成釉蛋白片段，我们检测了成骨细胞系MC3T3-E1经不同长度的成釉蛋白N末端区域处理后碱性磷酸酶的活性和钙化情况。结果表明，成釉蛋白(16N)N端的16个氨基酸(VPFFPQQSGTPGMASL)对细胞的诱导作用最强。为探讨16N在成骨细胞中的信号转导途径，观察16N对MC3T3-E1细胞MAPK表达的影响。细胞内可见p38的磷酸化。为了识别16N的受体，我们将编码16N的cDNA导入MC3T3-E1细胞。含有分泌序列的载体可诱导MC3T3-E1细胞产生较高的碱性磷酸酶活力，而不含分泌序列的载体则不能诱导细胞表达碱性磷酸酶。推测16N受体可能存在于MC3T3-E1细胞表面。尽管膜定位受体的鉴定尚未完成，但在MC3T3-E1细胞的细胞膜上检测到CD63，并怀疑其为16N.5的潜在受体。不同骨缺损模型的体内研究表明，16N具有促进再生的作用。这些结果表明，16N可以作为新型的骨愈合促进剂。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Studies on development of new regeneration therapy of bone by synthetic peptide of ameloblastin (in Japanese).

成釉素合成肽开发新型骨再生疗法的研究（日文）。

DOI：
发表时间：
2007
期刊：
影响因子：
0
作者：
Iizuka S;Yoshida M;Kitagawa M;Sakamoto K;Kawazoe Y;Miyauchi M;Takata T
通讯作者：
Takata T

Immortalization and characterization of human pulp cells with odontoblastic differentiation.

具有成牙本质细胞分化作用的人牙髓细胞的永生化和表征。

DOI：
发表时间：
2007
期刊：
Archives of Oral Biology 52
影响因子：
0
作者：
Sato S;Kitagawa M;Sakamoto K;Iizuka S;Kudo Y;Ogawa I;Miyauchi M;Foster BL;Somerman MJ;Takata T.;Masae Kitagawa
通讯作者：
Masae Kitagawa

Regeneration of bone by synthetic peptide of ameloblastin

成釉细胞合成肽促进骨再生