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

点击翻译按钮获取中文摘要

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