骨形成肽1和聚多巴胺复合涂层修饰提高聚醚醚酮表面活性

Modification with bone forming peptide 1 and polydopamine coating to improve bioactivity of polyetheretherketone surface

背景:聚醚醚酮具有与人体皮质骨相近的弹性模量及良好的射线透射性、化学稳定性和生物相容性等优点,有望应用于口腔种植领域,然而其具有生物惰性,难以与周围组织形成骨结合,因此如何提高聚醚醚酮的表面活性是目前的主要问题。目的:分析聚醚醚酮表面骨形成肽1和聚多巴胺复合涂层的促成骨和成血管作用。方法:将聚醚醚酮片浸泡于多巴胺溶液中24 h,制备聚醚醚酮-聚多巴胺材料;将聚醚醚酮-聚多巴胺材料浸泡于骨形成肽1溶液中24 h,制备聚醚醚酮-聚多巴胺-骨形成肽1材料,表征材料的微观形貌、亲水性与元素组成。将骨髓间充质干细胞分别接种于聚醚醚酮、聚醚醚酮-聚多巴胺、聚醚醚酮-聚多巴胺-骨形成肽1材料表面,通过活死细胞染色和细胞骨架染色评估细胞活性与黏附状态,茜素红和骨钙素免疫荧光染色检测细胞成骨分化能力。将人脐静脉内皮细胞分别接种于3组材料表面,通过活死细胞染色和细胞骨架/血管内皮生长因子免疫荧光染色评估细胞活性及成血管水平。结果与结论:①扫描电镜下可见聚醚醚酮材料表面光滑,聚醚醚酮-聚多巴胺材料表面出现凹凸不平的沉积物,聚醚醚酮-聚多巴胺-骨形成肽1材料表面有小颗粒突起;接触角测试结果显示,聚醚醚酮-聚多巴胺-骨形成肽1材料的亲水性优于其他两种材料;X射线光电子能谱测试结果显示,骨形成肽1成功修饰于聚醚醚酮材料表面;②活死细胞染色和细胞骨架染色显示,相较于其他两种材料,聚醚醚酮-聚多巴胺-骨形成肽1材料可提高骨髓间充质干细胞的活性与黏附;茜素红和骨钙素免疫荧光染色显示,相较于其他两种材料,聚醚醚酮-聚多巴胺-骨形成肽1材料可促进骨髓间充质干细胞的成骨分化;③活死细胞染色和免疫荧光染色显示,相较于其他两种材料,聚醚醚酮-聚多巴胺-骨形成肽1材料可提高人脐静脉内皮细胞的活性与黏附及血管内皮生长因子蛋白的表达;④结果表明,骨形成肽1和聚多巴胺复合涂层可提高聚醚醚酮表面的促成骨和成血管活性。

BACKGROUND:Polyether ether ketone has the advantages of elastic modulus close to human cortical bone,good ray transmission,chemical stability,and biocompatibility.Polyether ether ketone is expected to be applied in the field of oral implants.However,polyether ether ketone is bioinert and is difficult to integrate with surrounding bone tissue.Therefore,how to improve the surface bioactivity of polyether ether ketone remains a major issue.OBJECTIVE:To analyze the osteogenic and vasogenic effects of polyether ether ketone surface bone forming peptide 1 and polydopamine composite coating.METHODS:Polyether ether ketone titanium sheets were immersed in dopamine solution for 24 hours to prepare polyether ether ketone-polydopamine materials.Polyether ether ketone-polydopamine material was immersed in the bone forming peptide 1 solution for 24 hours to prepare polyether ether ketonepolydopamine-bone forming peptide 1 material.The micromorphology,hydrophilicity,and elemental composition of the material were characterized.Bone marrow mesenchymal stem cells were injected on the surface of polyether ether ketone,polyether ether ketone-polydopamine,and polyether ether ketonepolydopamine-bone forming peptide-1 materials.Cell viability and adhesion status were evaluated by live/dead cell staining and cytoskeleton staining.Cellular osteogenic differentiation ability was detected by alizarin red and osteocalcin immunofluorescence staining.Human umbilical vein endothelial cells were inoculated on the surface of three groups of materials.Cell viability and angiogenesis level were assessed by live/dead cell staining and immunofluorescence staining of cytoskeleton/vascular endothelial growth factor.RESULTS AND CONCLUSION:(1)Under scanning electron microscope,the surface of polyether ether ketone material was smooth;the surface of polyether ether ketone-polydopamine material had uneven deposits,and the surface of polyether ether ketone-polydopamine-bone forming peptide 1 material had small particles protruding.The results of contact angle test showed that the hydrophilicity of polyether ether ketone-polydopamine-bone forming peptide-1 material was better than the other two materials.X-ray photoelectron spectroscopy results showed that bone forming peptide 1 had been successfully modified on the surface of polyether ether ketone material.(2)Live/dead cell staining and cytoskeleton staining exhibited that compared with the other two materials,polyether ether ketone-polydopamine-bone-forming peptide 1 material could improve the viability and adhesion of bone marrow mesenchymal stem cells.Alizarin red and osteocalcin immunofluorescence staining showed that compared with the other two materials,polyether ether ketonepolydopamine-bone forming peptide-1 materials could promote the osteogenic differentiation of bone marrow mesenchymal stem cells.(3)Live/dead cell staining and immunofluorescence staining showed that compared with the other two materials,polyether ether ketone-polydopamine-bone forming peptide 1 material could improve the viability and adhesion of human umbilical vein endothelial cells and the expression of vascular endothelial growth factor protein.(4)The results show that the modification of bone forming peptide 1 and polydopamine composite coating on polyether ether ketone surface can improve the osteogenesis and angiogenesis.