二氧化钛纳米管材料成骨及抗菌实验研究

发布时间：2018-06-08 17:46

本文选题：钛纳米管 + 骨髓间充质干细胞　；参考：《第二军医大学》2013年博士论文

【摘要】：人工关节置换术后假体周围感染是人工关节置换术后灾难性的并发症，已经感染的人工关节及生物膜没有特殊有效的治疗方法，因此感染的预防非常重要。钛纳米管材料由于其纳米级的细微结构和规则的空间构造，具有优良的生物相容性同时也是天然的载药系统。因而通过钛合金内置物纳米管表面负载抗菌药物，可成为促进成骨长入，预防假体细菌粘附生长的有效方法，并可为预防治疗假体周围松动感染提供新的思路和选择。目的：首先通过电化学阳极氧化法制备出二氧化钛纳米管涂层并进行结构表征和电镜观察，控制不同的制备条件下制备不同纳米管孔径的材料并比较其结构特性。然后在钛纳米管材料表面进行骨髓间充质干细胞的培养，观察其粘附，生长和分化的活性，并在不同的纳米结构表面进行比较，探讨纳米管结构对成骨细胞的作用。而后在已制备的钛纳米管材料表面负载庆大霉素，进行载药和释药的研究，分别在体外和体内进行实验观察载药钛纳米管材料对金葡菌的抑菌作用。方法： 1.在氢氟酸和中性氟化物溶液中分别用电化学阳极氧化法制备钛纳米管材料。应用X射线能谱分析其表面元素，X射线衍射仪分析其晶体结构，电镜下观察和测量其结构特征，测量表面接触角和表面粗糙度。控制不同的电压制备不同纳米孔径的钛纳米管材料。 2.在钛纳米管材料表面培养骨髓间充质干细胞，应用MTT法观察其活性，荧光显微镜及电镜观察其形态，观察其粘附生长和分化情况。并与纯钛表面进行比较。尤其在不同孔径的钛纳米管材料表面进行干细胞的培养，观察其粘附和分化能力，分析其与钛纳米管表面结构的相互关系。 3.应用负压吸附法在钛纳米管材料表面负载庆大霉素，计算其药物负载率。通过高分子聚合物覆盖，观察其对药物释放曲线的影响。在体外实验中，比较载药和未载药的钛纳米管材料表面的金葡菌的粘附生长情况，并同样进行成骨细胞培养观察载药是否会对钛纳米管的促成骨作用产生影响。在体内实验中，通过在小鼠双下肢的对照实验，比较载庆大霉素纳米管材料在体内对金葡菌的抑制作用。结果：1.应用电化学阳极氧化法制备出钛纳米管材料，其在400°C热处理后主要为锐钛矿型，电镜下观察到其完整的纳米管形成的过程。控制不同的恒定电压可制备出不同孔径（随电压递增）的纳米管材料。 2.钛纳米管材料表面较纯钛对骨髓间充质细胞有显著的促进粘附，生长和分化的作用，不同孔径的钛纳米管材料表面对间充质干细胞的作用有所不同，管径小时促进其粘附和生长，管径大时干细胞形态变长，分化成骨能力增强。 3.钛纳米材料表面可有效负载庆大霉素，其释药曲线存在爆发性释放，在其表面覆盖高分子聚合物可有效延长其释放时间。体外实验证实载药钛纳米管材料可有效抑制金葡菌生长，并同时仍有促成骨作用。体内实验中载药钛纳米管可有效预防小鼠肢体感染。结论：1.电化学阳极氧化法可简单有效制备钛纳米管材料，，并可控制其表面结构参数。 2.钛纳米材料表面与成骨细胞系列有优良的生物相容性，促进材料表面成骨加强内植物骨融合稳定性。 3.钛纳米管材料表面可有效吸附抗菌药物，并具有有效的抗菌能力，对预防治疗假体周围感染具有良好的临床应用前景。
[Abstract]:The titanium nanotube material has excellent biocompatibility and is also a natural drug loading system due to the fine structure and the regular space structure of the titanium alloy built - in nanotube surface , thereby being an effective method for promoting the bone ingrowth and preventing the adhesion and growth of the prosthesis bacteria , and can provide a new idea and selection for preventing loosening infection around the prosthesis .

Objective : To prepare the titanium dioxide nanotube coating by electrochemical anodic oxidation and to observe its structural characteristics and to compare its structural properties under different preparation conditions . Then , the effects of nanotube structure on osteoblast were observed under different preparation conditions . Then , gentamicin was loaded on the surface of titanium nanotube material .

Method :

1 . The titanium nanotube material was prepared by electrochemical anodic oxidation method in hydrofluoric acid and neutral fluoride solution . The surface element was analyzed by X - ray energy dispersive X - ray diffraction . The crystal structure was analyzed by X - ray diffraction and its structural features were observed and measured under electron microscope . The surface contact angle and surface roughness were measured .

2 . The bone marrow mesenchymal stem cells were cultured on the surface of titanium nanotube material . The morphology of the bone marrow mesenchymal stem cells was observed by MTT assay . The adherent growth and differentiation were observed .

3 . Using negative pressure adsorption method to load gentamicin on the surface of titanium nanotube material , the drug loading rate was calculated .

Results : 1 . The titanium nanotube material was prepared by electrochemical anodic oxidation method , which was mainly anatase type after heat treatment at 400 掳 C . The process of its complete nanotube formation was observed under electron microscope . Different constant voltage could be controlled to prepare nanotube material with different pore size ( increasing with voltage ) .

2 . The surface of the titanium nanotube material has a significant effect on the adhesion , growth and differentiation of the mesenchymal stem cells , and the surface of the titanium nanotube material with different pore sizes has different effects on the mesenchymal stem cells , and the diameter of the titanium nanotube material can promote the adhesion and growth of the mesenchymal stem cells , and the stem cell morphology becomes longer when the diameter is large , and the differentiation into the bone capacity is enhanced .

3 . The surface of the titanium nano - material can be effectively loaded with gentamicin , and the drug release curve of the titanium nano - material can effectively prolong the release time of the drug - loaded titanium nanotube material .

Conclusion : 1 . Electrochemical anodic oxidation method is simple and effective to prepare titanium nanotube material , and its surface structure parameters can be controlled .

2 . The surface of the titanium nano material has excellent biocompatibility with the osteoblast series , and the surface of the material is promoted to strengthen the fusion stability of the bone in the internal plant .

3 . The surface of the titanium nanotube material can effectively adsorb the antibacterial drugs , and has effective antibacterial ability , and has good clinical application prospect for preventing the periprosthetic infection .
【学位授予单位】：第二军医大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R318.08;TB383.1

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