【作者】 刘伟；

【导师】 张先龙；

【作者基本信息】 上海交通大学 ， 外科学（骨外）， 2019， 博士

【摘要】 聚醚醚酮(PEEK)由于其优异的力学性能,被认为是最有前景的骨科内植物之一。但是其生物相容性及骨整合性较差,限制其在临床的广泛使用。虽然前期有研究证明了通过磺化技术可以在PEEK表面制备多孔结构,一定程度提高PEEK的生物相容性,但其成骨活性改善却不明显。因此,本研究拟在表面多孔PEEK上通过磁控溅射技术添加促成骨及免疫调节的锌元素,从而同时从直接成骨及免疫成骨两个方面提高PEEK的成骨活性,并且研究锌元素在免疫成骨方面的效应。我们使用磺化技术联合磁控溅射制备掺锌表面多孔PEEK(Zn-SPEEK)。材料表征检测方面,我们利用扫描电镜及能谱仪检测Zn-SPEEK表面形貌及元素分布情况;原子力学显微镜检测材料表面3D形貌;X射线衍射仪检测锌元素的价态。使用细胞电镜,增殖实验以及流式细胞仪检测材料的生物相容性。体外免疫调节方面检测,我们首先通过RNA-seq技术掌握巨噬细胞在材料表面基因表达情况,之后通过免疫荧光,ELISA及流式细胞技术等分析巨噬细胞亚型。通过碱性磷酸酶及茜素红染色,免疫荧光及RT-PCR技术检测Zn-SPEEK的免疫成骨能力,最后分别通过气囊模型及体内骨缺损模型来验掺锌表面多孔PEEK的体内免疫调节及骨修复能力。结果显示,利用磺化及磁控溅射技术可以制备掺锌表面多孔PEEK,Zn-SPEEK拥有良好的生物相容性,体外及体内实验的结果均提示Zn-SPEEK可以诱导巨噬细胞向M2进行表型转化,进而分泌促成骨的细胞因子,其诱导的免疫微环境有利于骨髓间充质干细胞的成骨分化,从而提高PEEK的骨整合能力。由此,本研究结论为经过磺化及磁控溅射制备的Zn-SPEEK不仅具有良好的生物相容性,而且可以通过调节免疫促进骨修复,是一种有前景的骨科内植物材料。更多还原

【Abstract】 Polytheretherketone(PEEK)is thought to be one of the most promising orthopedic implants due to its excellent mechanical properties.However,poor biocompatibility and osteointegration hinder its widely clinical application.Some studies have reported that a surface porous coating obtained by sulfonation on PEEK can improved the biocompatibility,but the osteointegration of PEEK was not enhanced significantly.Herein,we incorporated zinc,an osteogenic and immunomodulatory element,on the surface of surface porous PEEK through magnetron sputtering to improve the osteointegration of PEEK,and further investigated the immunomodulatory effect of zinc.Zinc-incorporated surface porous PEEK(Zn-SPEEK)was successfully prepared through sulfonation and magnetron sputtering.Scanning electron microscopy(SEM)and energy dispersive spectrometer(EDS)were used to determine the surface morphology and element distribution of Zn-SPEEK.Atom force microscopy and X-ray photoelectron spectroscopy were employed to detect the 3D morphology and element valence of Zn-SPEEK.Then the cell biocompatibility was evaluated by SEM,cell viability test and flow cytometer.Regarding the immunomodulatory effect of Zn-SPEEK,we first examine the gene expression profile of macrophage cultured on samples using RNA-seq.Immunofluorescence,flow cytometer and ELISA were used to determine the subtype of macrophages and its cytokines.To acquire the immunomodulatory osteogenic effect of Zn-SPEEK,we performed alkaline phosphatase(ALP)and alizarin red staining,immunofluorescence and RT-PCR tests.Finally,the in vivo air pouch model and bone defect repair model were built to validate the immunomodulatory and osteogenic activity of Zn-SPEEK.Our results indicated that Zn-SPEEK showed good biocompatibility and can modulate macrophage polarizing to M2 phenotype,secreting osteogenic cytokines to generate a microenvironment favoring osteogenesis both in vitro and vivo.Therefore,we concluded that zinc-incorporated surface porous PEEK(Zn-SPEEK)can be prepared through sulfonation and magnetron sputtering.The favorable biocompatibility and immunomodulatory osteogenic effect of Zn-SPEEK pave the way for its further application as orthopedic implants.更多还原