基于聚多巴胺的材料表面修饰对二磷酸腺苷促血小板聚集功能的影响

摘要

目的:采用基于聚多巴胺的表面改性方法,将具有促血小板聚集作用的二磷酸腺苷接枝到材料表面,构建一种具有促进血小板黏附、聚集与激活表面.方法:通过聚多巴胺偶联作用将二磷酸腺苷接枝到细胞爬片表面(G-ADP组),另设两个对照组分别为多巴胺涂层细胞爬片表面(G-DA组)和细胞爬片表面(G组).通过X射线光电子能谱分析材料表面化学元素;在材料表面培养人骨髓间充质干细胞,检测1、3、5天细胞增殖情况;通过场发射扫描电镜、乳酸脱氢酶定量实验和免疫荧光标记实验测定材料表面血小板黏附、聚集与激活情况.结果:G-ADP表面出现P峰,表明二磷酸腺苷接枝到材料表面;三组材料表面培养细胞均呈增殖状态,且三组之间差异无统计学意义P＞0.05.电镜下观察G-ADP表面黏附大量血小板,聚集成簇,伸出伪足并且相互融合,而对照组仅有少量血小板黏附;定量实验显示G-ADP表面血小板黏附量高于其它两组;G-ADP表面CD41-FITC和CD62P-PE两种抗体的荧光强度均显著高于其它两组.结论:二磷酸腺苷可成功接枝到材料表面,功能化修饰表面可促进血小板黏附、聚集与激活,且具有良好的生物相容性.%Objective:To establish a surface that promote platelet adhesion,aggregation and activation by grafting adenosine diphosphate to the material surface and have good biocompatibility.Methods:Adenosine diphosphate was grafted onto the glass surface by coupling effect of polydopamine (G-ADP group).Dopamine coated glass surface (G-DA group) and uncoated glass surface (G group) were two control groups.X-ray photoelectron spectroscopy was performed to analyze surface chemical element.Human Bone Mesenchymal Stem Cells were cultured on surfaces and determined the proliferation of cells on 1,3,5 day.Field Emission Scanning Electron Microscope,lactate dehydrogenase quantitative assay and immunofluorescence marker assay were performed to demonstrate the amount,morphology and activity of the adhered platelets.Results:XPS showed that G-ADP surface appeared P peaks indicating that adenosine diphosphate was grafted to the surface.The number of cells in three groups rose respectively with no significant differences (P＞0.05).Under electron microscope,G-ADP surface was covered by a large number of platelets gathered in clusters,extended pseudopodia and fused mutually,while the control groups weren't.The adhesive amount of platelets on G-ADP surface was significantly higher than the others quantitatively.Fluorescence intensity of CD41-FITC and CD62P-PE in G-ADP group were both significantly higher than the other two groups.Conclusion:Adenosine diphosphate can be successfully grafted onto the surface.Functionalized surface can promote adhesion,aggregation and activation ofplatelets with good biocompatibility.