纤维连接蛋白模拟多肽改进碳磷酸钙骨水泥反应性的体外研究

Enhancement of the osteoblast on calcium carbonated phosphate cement surface with fibronectin like peptides-an in vitro study

目的:寻找改进羟基磷灰石类骨修复材料生物活性的方法。方法:用固相法合成了多肽P19和P18,测定了P19与羟基磷灰石的结合特性,测定了P19对于成骨细胞附着的影响以及在P18与P19存在的条件下,成骨细胞的增殖特性,并用扫描电镜观察了附着在碳磷酸钙骨水泥(calciumcarbonatedphosphatecement,CPCC)材料表面的成骨细胞形态,并用DNA测量法测定了细胞在材料表面增殖特性。结果:P19铺板后能增强P18促进细胞增殖的作用。多肽分子P19对于成骨细胞的作用与纤维连接蛋白类似,多肽和CPCC结合后仍然保持其各自的生物活性,吸附在材料表面的P19能够促进成骨细胞在材料表面的附着,经过P19多肽修饰的CPCC表面能够明显增加成骨细胞吸附的数量,并能够促进成骨细胞的增殖。P18和P19之间存在一定的协同作用。结论:合成的P19融合肽可以作为HA类骨损伤修复材料的生物活性添加成分以及种植体涂层的修饰,这一方法和技术在HA骨修复材料的改进以及种植体涂层修饰方面有着潜在的临床应用前景。

Objective:To explore the methods to improve the bioactive reactivity of calcium carbonated phosphate cement(CPCC). Methods:Peptides named P19 and P18 were designed and synthesized by the method of solid phase synthesis. The proliferation of osteoblast under existence of P18 and P19 was investigated. Scan electron microscope and DNA fluo rescent assay was used to observe the shape and amount of osteoblast attached on CPCC disc. The synergistic effect of P19 and p18 was also explored. Results:P19 increased the amount of attached osteoblast on cell culture plate surface and en hance the proliferation of attached cell whether used alone or together with P18. Osteoblast pretreated with P19 can not at tached to the culture plate surface, which was coated with fibronectin. CPCC disc surface modified with P19 not only obvi ously promote d the attachment osteoblast cell, but also enhancd proliferation of osteoblast cell.P18 synergistically affected the proliferation of osteoblast with P19. Conclusion:Synthesized peptides can be used as biological active modifier of hy droxyapatite based materials by in vitro study. Furthermore, the technology of synthesized peptides and used in our study may have wide clinical application prospect in the area of hydroxyapatite based material improvement and modification of dental implant coating layer.