银离子注入钛的抗菌、成骨作用研究

发布时间：2018-05-20 10:15

本文选题：生物膜 + 内植物感染　；参考：《上海交通大学》2015年博士论文

【摘要】：细菌在内植物材料表面发生粘附、繁殖,形成生物膜,引起内植物感染是临床上导致内植物固定失败的一个主要原因。内植物缺乏骨整合性能是另一个引起失败的原因。而目前常用的内植物材料——钛缺乏抗菌能力和骨整合性能。本研究通过等离子体浸没、银离子注入(silver plasma immersion ion implantation,Ag-PIII)技术对钛进行表面改性,在钛表面植入纳米银;重点探讨改性后的钛的抗生物膜性能及骨整合性能。通过Ag-PIII技术,我们对钛表面进行改性。通过场发射扫描电镜观察钛的表面形态;通过X射线光电子谱检测钛表面的元素价态;通过电感耦合等离子体光学发射光谱法检测改性后钛在电解质液体中银的释放。观察细菌在材料表面的生长状况,通过活菌涂板计数法、扫描电镜、激光共聚焦显微镜等方法,检测改性后钛的抑制生物膜的能力。采用与内植物相关的骨髓炎动物模型,在体内验证材料的抗菌作用。使用人骨髓干细胞,通过体外检测细胞在材料表面的粘附、增殖、形态检测改性后钛的细胞相容性;通过检测碱性磷酸酶(ALP)活力、细胞外基质矿化能力、成骨相关基因表达来验证材料的促成骨能力;并通过检测了细胞的Rho A蛋白的水平来验证可能的细胞通路。采用动物模型体内验证材料的骨整合作用。结果表明,通过Ag-PIII技术,可以在钛表面定植纳米银;定植纳米银的量与Ag-PIII时间成正比;并且在电解质液体中释放极微量的银离子。定植的纳米银抑制了细菌在钛表面形成生物膜,降低了内植物感染的发生率。并且没有明显的细胞毒性作用,提高了骨髓干细胞的早期粘附能力,增加了ALP活力、细胞外基质矿化水平、上调了成骨相关基因的表达水平,提高了GTP-Rho A蛋白的表达;改善了钛的骨整合能力。由此我们得出结论,通过Ag-PIII技术,我们可以在钛表面可控的定植纳米银,赋予钛抗生物膜和骨整合能力。
[Abstract]:Adhesion, reproduction and biofilm formation of bacteria on the surface of plant materials are one of the main reasons for the failure of endophyte fixation. The lack of bone integration in implants is another cause of failure. Titanium, a commonly used implant material, lacks antimicrobial activity and bone integration. In this study, the surface modification of titanium was carried out by plasma immersion and silver ion implantation with silver plasma immersion ion implantationsAg-PIII. The anti-biofilm and bone integration properties of the modified titanium were investigated. The surface of titanium was modified by Ag-PIII technology. The surface morphology of titanium was observed by field emission scanning electron microscope (SEM), the valence state of titanium surface was detected by X-ray photoelectron spectroscopy (XPS), and the silver release in electrolyte liquid was detected by inductively coupled plasma optical emission spectrometry (ICP-OAES). The growth of bacteria on the surface of the material was observed. The ability of the modified titanium to inhibit biofilm was tested by means of counting method of live bacteria coating, scanning electron microscope, laser confocal microscope and so on. An animal model of osteomyelitis related to implants was used to verify the antibacterial effect of the material in vivo. Using human bone marrow stem cells, the cell adhesion, proliferation and morphology of the modified titanium were detected in vitro, and the activity of alkaline phosphatase (ALP) and the mineralizing ability of extracellular matrix were detected. Osteoblast-associated gene expression was used to verify the ability of the material to promote bone, and the level of Rho A protein was detected to verify the possible cellular pathway. Animal models were used to verify the osteointegration of the materials. The results show that nanocrystalline silver can be grown on titanium surface by Ag-PIII technique, the amount of silver can be directly proportional to the time of Ag-PIII, and very small amount of silver ions can be released in electrolyte liquid. The bacterial biofilm formation on titanium surface was inhibited and the incidence of endophytic infection was decreased. Moreover, there was no obvious cytotoxic effect, which increased the ability of bone marrow stem cells to adhere early, increased the activity of ALP, mineralization of extracellular matrix, up-regulated the expression of osteoblast related genes, and increased the expression of GTP-Rho A protein. The bone integration ability of titanium was improved. From this we conclude that through Ag-PIII technology we can implant nano-silver on titanium surface and endow titanium with anti-biofilm and bone integration ability.
【学位授予单位】：上海交通大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R318.08;R687

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