牙本质基质蛋白—1仿生多肽设计与评价.docVIP

牙本质基质蛋白—1仿生多肽设计与评价 　　[摘要] 目的 设计一种牙本质基质蛋白-1（DMP-1）的仿生多肽，仿生DMP-1与胶原纤维结合，并启动矿化功能。 　　方法 依据DMP-1与胶原纤维结合的功能序列以及釉原蛋白介导羟磷灰石成核的功能序列，采用固相合成法合成DMP-1仿生多肽，其氨基酸序列为DSESSEEDRTKREEVD。利用免疫荧光法评价该多肽与胶原蛋白的结合能力；将其依次浸入氯化钙溶液和磷酸氢二钠溶液中，采用扫描电子显微镜（SEM）和透射电子显微镜（TEM）以及选区电子衍射环（SAD）分析其诱导羟磷灰石晶体成核生长的矿化能力。结果 免疫荧光结果表明：本研究设计的DMP-1仿生多肽可以与牙本质胶原纤维结合；SEM、TEM观察结果显示：该多肽可以从溶液中摄取钙磷离子，诱导羟磷灰石晶体成核矿化。结论 多肽DSESSEEDRTKREEVD可以模拟DMP-1与胶原蛋白结合及启动矿化的能力，可以作为研究牙本质仿生矿化的一种分子工具。 　　[关键词] 牙本质基质蛋白-1； 多肽； 仿生； 生物矿化 　　[中图分类号] R 739.86 [文献标志码] A [doi] 10.7518/hxkq.2013.04.003 　　Design and evaluation of a kind of biomimetic peptides of dentin matrix protein-1 Liu Wei1， Cao Ying1，2， Shen Jun3， Xu Qiangjian1， Zhu Zhenxiong2， Li Quanli1. （1. Dept. of Prosthodontics， Stomatologic Hospital College of Anhui Medical University， Key Laboratory of Oral Diseases Research of Anhui Province， Hefei 230032， China； 2. Fa-culty of Dentistry， The University of Hong Kong， Hong Kong 810000， China； 3. Dept. of Orthodontics， Stomatologic Hospital College of Anhui Medical University， Key Laboratory of Oral Diseases Research of Anhui Province， He-fei 230032， China） 　　[Abstract] Objective To design a kind of biomimetic polypeptide of dentin matrix protein-1（DMP-1）， which can 　　bind to dentine collagen fibers and initiate mineralization. Methods A novel polypeptide was developed by connecting the collagen binding domain of DMP-1 “DSESSEEDR” to the hydrophilic C-terminal of amelogenin “TKREEVD”. The polypeptide was synthetically prepared by standard solid-phase peptide synthesis. Human dentine slices were completely demineralized by hydrochloric acid to expose the dentine collagen. Fluorescein isothiocyanate coupled polypeptide was applied to the exposed dentine collagen. Fluorescent microscopy was used to examine the polypeptide specially bond to the dentine collagen. Nucleation and growth of hydroxyapatite was initiated by immersing the polypeptide into cal-cium chloride and sodium hypophosphate solutions respectively. Scanning electron microscopy