两种抗菌性纳米网层钛材料的抗菌活性及生物相容性的初步研究

发布时间：2018-02-12 06:51

本文关键词： 纳米网层钛材料紫外光处理纳米银粒子抗菌活性大鼠骨髓间充质干细胞　出处：《南方医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：研究背景和目的:钛材料由于其能够与人体骨组织产生骨结合,以及具有优良的机械性能和抗腐蚀性能,被广泛应用于口腔的临床治疗。然而,无论是短期的种植体手术术后感染,还是长期的种植体周围炎,均可能导致种植治疗的失败。因此,赋予钛种植体抗菌性是一种预防种植体周感染、提高种植术成功率的方法。高浓度碱处理能够在钛材料表面形成疏松多孔的纳米级网层状结构,被称为纳米网层钛材料。此材料虽然拥有优良的促成骨分化能力,但缺乏对细菌入侵的抵抗能力。UV处理钛材料不仅能够带来更快更完整的骨结合,还能够减少细菌早期附着。另外,以不同方式加载纳米银粒子的钛材料也显示了长效的抗菌性和较好的细胞相容性。本研究用UV处理和加载纳米银粒子两种方式,制备两种具有不同时效性的抗菌性钛材料,并评价其抗菌能力以及对大鼠骨髓间充质干细胞的影响。方法:经过研磨的纯钛片浸泡于10 M NaOH溶液中室温处理24小时后,用高强度短时间的UV照射制备成UV处理纳米网层钛材料,以及硝酸银溶液浸泡UV照射的方式在纳米网层钛材料表面加载纳米银离子。运用扫描电镜(SEM)、X射线光电子能谱分析(XPS)、接触角测量等方法,分析两种纳米网层钛材料的表面特征。以龈下菌斑早期定植细菌Actinomycesoris MG-1作为菌种评价两种纳米网层钛材料的抗菌能力。另外,在两种抗菌性纳米网层钛材料表面培养大鼠骨髓间充质干细胞以分析其成骨分化活性。研究结果:1、高强度短时间UV照射使纳米网层钛材料获得超亲水性,并有效去除表面碳污染;以硝酸银溶液浸泡UV照射还原的方式,成功制备加载纳米银离子纳米网层钛材料。2、UV处理纳米网层钛材料有效地抵抗Actinomycesoris MG-1的早期附着,并且抑制其生物膜形成达6小时。3、载银纳米网层钛材料不仅能够杀灭已附着细菌,抑制生物膜形成,并且显示出长效抗菌性达28天。4、UV处理纳米网层钛材料促进大鼠骨髓间充质干细胞的早期附着与增殖、ALP活性、骨钙素分泌以及细胞外基质矿化,成骨相关基因(ALP、Runx2、BMP、OPN)表达水平均高于纳米网层钛材料。5、载银纳米网层钛材料与纳米网层钛材料相比,抑制大鼠骨髓间充质干细胞的早期附着与增殖以及ALP活性。结论:UV处理纳米网层钛材料能够抗菌达6小时以上,并且能够促进大鼠骨髓间充质干细胞的成骨分化,载银纳米网层钛材料保持抗菌性达28天,但有一定程度的细胞毒性。本研究将进一步深入探讨增强载银纳米网层钛材料的生物相容性。
[Abstract]:Background and objective: titanium is widely used in clinical treatment of oral cavity due to its ability to bind bone to human bone tissue, as well as its excellent mechanical properties and corrosion resistance. Both short-term post-implant infections and long-term peri-implant infections can lead to failure in implant therapy. Therefore, giving titanium implants antimicrobial properties is a way to prevent peri-implant infection. The method of improving the success rate of implantation. High concentration alkali treatment can form porous and porous nano-mesh layered structure on the surface of titanium, which is called nano-mesh layer titanium material, although this material has excellent ability to promote bone differentiation. But the lack of resistance to bacterial invasion. UV treatment of titanium not only leads to faster and more complete bone bonding, but also reduces the early attachment of bacteria. Titanium materials loaded with silver nanoparticles in different ways also showed long-term antibacterial properties and good cytocompatibility. In this study, two kinds of antimicrobial titanium materials with different time-efficiency were prepared by UV treatment and loading of nano-silver particles. To evaluate its antibacterial ability and its effect on rat bone marrow mesenchymal stem cells. Methods: after being soaked in 10 M NaOH solution at room temperature for 24 hours after grinding pure titanium slices. Titanium nanowires were prepared by UV irradiation with high intensity and short time. The silver nanoparticles were loaded on the surface of titanium nanowire by soaking in silver nitrate solution. The X-ray photoelectron spectroscopy (XPS) analysis and contact angle measurement were used to analyze the silver ions on the surface of titanium nanowet layer by means of scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and contact angle measurement, etc. The surface characteristics of two kinds of nanometer-netted titanium materials were analyzed. The antibacterial ability of the two nano-netted titanium materials was evaluated by early colonization bacteria (Actinomycesoris MG-1) in subgingival plaque. Rat bone marrow mesenchymal stem cells (BMSCs) were cultured on the surface of two kinds of antibacterial nano-netting titanium materials to analyze their osteogenic differentiation activity. The surface carbon pollution was removed effectively, and the loaded nano-silver ion nanowire layer titanium material. 2 ~ (2 +) UV was successfully prepared by immersing in silver nitrate solution to UV irradiation and reducing, and effectively resisting the early attachment of Actinomycesoris MG-1 to the nano-netted titanium material treated by UV. Moreover, the formation of biofilm was inhibited for 6 hours. The silver loaded nanofilm layer titanium not only killed the attached bacteria, but also inhibited the formation of biofilm. The results showed that the long-term antibacterial activity was 28 days. 4 days after treatment with nanowet layer titanium, the early attachment and proliferation of rat bone marrow mesenchymal stem cells (BMSCs) and ALP activity, osteocalcin secretion and extracellular matrix mineralization were promoted. The expression level of osteoblast-associated gene (ALP) Runx 2 (BMPN) was significantly higher than that of nano-mesh layer titanium (0.05), and that of silver-loaded nano-mesh layer titanium was higher than that of nano-netted titanium material. The inhibition of bone marrow mesenchymal stem cells (BMSCs) in the early stage of attachment and proliferation and the activity of ALP. Conclusion the treatment of nanometer-net layer titanium can inhibit the activity of bone marrow mesenchymal stem cells for more than 6 hours and promote the differentiation of bone marrow mesenchymal stem cells. In this study, the biocompatibility of silver-loaded nanometer-net layer titanium material was further studied, which maintained antibacterial activity for 28 days, but had a certain degree of cytotoxicity.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R783.1

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