基于生物材料表界面调控的凝血-免疫级联过程研究

        骨缺损修复是一个复杂的、多细胞参与的级联过程，凝血与炎症作为植入物表面最初触发的生物反应，通过调控植入物的表面特性，可以实现植入体促修复功能的显著提升。然而，材料表/界面对早期血肿和炎症级联过程的影响及其调控机理仍有待深入探索。 
        因此，本论文聚焦于材料表面亲水性和粗糙度两大关键因素，探究材料表面特性对早期凝血与炎症反应的影响。本论文结合微球自组装技术和等离子体刻蚀技术，提高了聚醚醚酮表面亲水性，并通过构建表面锥刺状微纳结构实现了对表面粗糙度的调节。进一步构建体外血肿模型，通过扫描电子显微镜证明通过调控亲水性和粗糙度可以实现对血肿三维结构和组成的调节；结合聚合酶链式反应和酶联免疫吸附试验揭示了提高亲水性可以减弱血液中蛋白和相关细胞的粘附，进而血肿块中纤维蛋白直径降低、免疫细胞活性增强、炎症反应增强，而粗糙度增加可减弱以上影响。同时以巨噬细胞为炎症细胞代表，通过材料直接调控和血肿浸提液条件培养，结合聚合酶链式反应、酶联免疫吸附试验和免疫荧光染色等方法，证明了提高亲水性有利于巨噬细胞活化，而粗糙度的增强可减弱其炎症反应。综上所述，本论文揭示了材料表面性质在调控凝血与炎症反应中的重要作用，有望为医用植入材料的设计优化提供一定的理论支撑。

Bone defect repair is a complex, multicellular cascade process, and coagulation and inflammation, as biological responses initially triggered on the surface of the implant, can achieve a significant enhancement of the pro-repair function of the implant by modulating the surface properties of the implant. However, the influence of the material surface/interface on the early hematoma and inflammation cascade process and its regulatory mechanism still need to be deeply explored. 

Therefore, this dissertation focuses on two key factors, hydrophilicity and roughness of the material surface, to investigate the influence of material surface properties on early coagulation and inflammatory responses. In this thesis, the hydrophilicity of poly(ether ether ketone) surface was improved by combining microsphere self-assembly technology and plasma etching technology, and the modulation of surface roughness was realized by constructing surface cone-spine-like micro-nanostructures. Further, an in vitro hematoma model was constructed, and it was demonstrated by scanning electron microscopy that the regulation of hydrophilicity and roughness could realize the regulation of the three-dimensional structure and composition of the hematoma; the combination of polymerase chain reaction and enzyme-linked immunosorbent assay revealed that the increase of hydrophilicity could attenuate the adhesion of the blood 
proteins and the related cells, and consequently, the diameter of the fibrin in the  hematoma mass was reduced, the immune cells' activity was enhanced, and the  inflammatory response was strengthened, and the increase of roughness could attenuate the above mentioned effects. Increased roughness attenuates the above effects. Meanwhile, using macrophages as the representative of inflammatory cells, it was demonstrated that increased hydrophilicity favored macrophage activation, while enhanced roughness attenuated its inflammatory response through direct material modulation and hematoma extract conditioned culture, combined with polymerase chain reaction, enzyme-linked immunosorbent assay and immunofluorescence staining. In summary, this thesis reveals the important role of material surface properties in regulating coagulation and inflammatory responses, which is expected to provide certain theoretical support for the design optimization of medical implant materials. 

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