VEGF功能化镍钛合金对内皮细胞行为的影响
发布时间：2018-09-10 19:40
【摘要】：NiTi合金由于其独特的形状记忆效应、超弹性、高耐腐蚀性和良好的生物相容性等性能，已在生物医学领域得到广泛应用。然而，NiTi合金植入材料表面容易诱导血栓形成，是其临床应用中面临的主要挑战之一。此外，NiTi合金植入后在组织周围释放Ni2+可能会产生过敏反应和毒副作用。因此，通过NiTi合金基材的表面工程促进血管内皮细胞的功能（血管内皮化）同时减少Ni2+释放成为NiTi合金植入材料的研究重点。 研究表明：减少Ni2+释放量的一种有效方法是在NiTi合金表面形成一层保护涂层，，且该涂层可提高NiTi合金植入物的生物相容性，进而调控内皮细胞的生理功能。众所周知，细胞微环境在调节细胞行为方面发挥着重要作用，细胞行为（如黏附、迁移、增殖、分化等）都受到材料表面的化学成分和形貌的影响。在材料表面固定细胞外基质成分或生长因子可以改变材料表面的化学组成，从而促进细胞与生物材料相互作用，增强细胞的功能。内皮细胞生长因子（VEGF）可以促进血管内皮细胞的迁移、增殖并形成管状结构，从而促进血管的新生。 本文首先采用磁控溅射和水热氧化方法在NiTi合金表面构建纳米氧化铝涂层，然后以多巴胺为中间界层在其表面固定VEGF。利用X射线衍射（XRD）、扫描电镜（SEM）、原子力扫描电镜（AFM）、电感耦合等离子体质谱仪（ICP-MS）和X-射线光电子能谱仪（XPS）对相关材料进行了表征。材料表面为无定型的多孔的针状纳米结构氧化铝涂层，其均方根粗糙度为22±4nm，厚度大约为200nm。该涂层显著降低了材料表面的Ni2+释放量。同时，我们对人脐静脉内皮细胞在材料表面的细胞增殖、细胞形貌、细胞铺展以及细胞分泌的前列环素I2（PGI2）和一氧化氮（NO）进行了检测。结果表明：接种在VEGF修饰后的NiTi合金表面的人脐静脉内皮细胞与纳米涂层的NiTi合金以及未修饰的NiTi合金相比，具有更高的活力(p 0.05或p 0.01)，并且内皮细胞在VEGF修饰后NiTi合金表面的数量更多，铺展面积更大。同时，内皮细胞在VEGF修饰后NiTi合金表面PGI2以及NO的分泌量与未修饰的NiTi合金相比明显增加，均具有显著性差异性（p 0.05）。本研究为NiTi合金在生物医学领域的广泛应用积累了有益的实验数据。
[Abstract]:NiTi alloys have been widely used in biomedical fields due to their unique shape memory effect, super elasticity, high corrosion resistance and good biocompatibility. However, the surface of Niti alloy implant is easy to induce thrombosis, which is one of the main challenges in clinical application. In addition, the release of Ni2 around the tissue may result in allergic reactions and toxic side effects after implantation of Niti alloy. Therefore, the surface engineering of NiTi alloy substrate can promote the function of vascular endothelial cells (vascular endothelialization) and reduce the release of Ni2. The results showed that an effective way to reduce the release of Ni2 was to form a protective coating on the surface of NiTi alloy, which could improve the biocompatibility of NiTi alloy implants and regulate the physiological function of endothelial cells. It is well known that cell microenvironment plays an important role in regulating cell behavior. Cell behavior (such as adhesion migration proliferation differentiation and so on) is affected by the chemical composition and morphology of the material surface. Immobilization of extracellular matrix or growth factor on the surface of the material can change the chemical composition of the material surface, thus promote the interaction between cells and biomaterials and enhance the function of cells. Endothelial growth factor (VEGF) can promote vascular endothelial cell migration, proliferation and formation of tubular structure, thereby promoting angiogenesis. In this paper, nanocrystalline Al _ 2O _ 3 coating was prepared on NiTi alloy by magnetron sputtering and hydrothermal oxidation. Then VEGF. was fixed on the surface of NiTi alloy with dopamine as the intermediate layer. The related materials were characterized by X-ray diffraction (XRD), scanning electron microscope (XRD), (SEM), atomic force scanning electron microscope (AFM), inductively coupled plasma mass spectrometer (ICP-MS) and X-ray photoelectron spectroscopy (XPS). The surface of the material is an amorphous porous needle-like nano-structure alumina coating with a root-mean-square roughness of 22 卤4 nm and a thickness of about 200 nm. The Ni2 emission on the surface of the material was significantly reduced by the coating. At the same time, the cell proliferation, cell morphology, cell spreading, prostacyclin I 2 (PGI2) and nitric oxide (NO) (no (NO) of human umbilical vein endothelial cells on the surface of materials were detected. The results showed that the human umbilical vein endothelial cells inoculated on the surface of VEGF modified NiTi alloy were compared with nano-coated NiTi alloy and unmodified NiTi alloy. It has higher activity (p0.05 or p0.01), and the number of endothelial cells on the surface of VEGF modified NiTi alloy is more and the spreading area is larger. At the same time, the secretion of PGI2 and NO on the surface of VEGF modified NiTi alloy was significantly higher than that of unmodified NiTi alloy (p0.05). This study has accumulated useful experimental data for the wide application of NiTi alloy in biomedical field.
【学位授予单位】：重庆大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.08
本文编号：2235422
[Abstract]:NiTi alloys have been widely used in biomedical fields due to their unique shape memory effect, super elasticity, high corrosion resistance and good biocompatibility. However, the surface of Niti alloy implant is easy to induce thrombosis, which is one of the main challenges in clinical application. In addition, the release of Ni2 around the tissue may result in allergic reactions and toxic side effects after implantation of Niti alloy. Therefore, the surface engineering of NiTi alloy substrate can promote the function of vascular endothelial cells (vascular endothelialization) and reduce the release of Ni2. The results showed that an effective way to reduce the release of Ni2 was to form a protective coating on the surface of NiTi alloy, which could improve the biocompatibility of NiTi alloy implants and regulate the physiological function of endothelial cells. It is well known that cell microenvironment plays an important role in regulating cell behavior. Cell behavior (such as adhesion migration proliferation differentiation and so on) is affected by the chemical composition and morphology of the material surface. Immobilization of extracellular matrix or growth factor on the surface of the material can change the chemical composition of the material surface, thus promote the interaction between cells and biomaterials and enhance the function of cells. Endothelial growth factor (VEGF) can promote vascular endothelial cell migration, proliferation and formation of tubular structure, thereby promoting angiogenesis. In this paper, nanocrystalline Al _ 2O _ 3 coating was prepared on NiTi alloy by magnetron sputtering and hydrothermal oxidation. Then VEGF. was fixed on the surface of NiTi alloy with dopamine as the intermediate layer. The related materials were characterized by X-ray diffraction (XRD), scanning electron microscope (XRD), (SEM), atomic force scanning electron microscope (AFM), inductively coupled plasma mass spectrometer (ICP-MS) and X-ray photoelectron spectroscopy (XPS). The surface of the material is an amorphous porous needle-like nano-structure alumina coating with a root-mean-square roughness of 22 卤4 nm and a thickness of about 200 nm. The Ni2 emission on the surface of the material was significantly reduced by the coating. At the same time, the cell proliferation, cell morphology, cell spreading, prostacyclin I 2 (PGI2) and nitric oxide (NO) (no (NO) of human umbilical vein endothelial cells on the surface of materials were detected. The results showed that the human umbilical vein endothelial cells inoculated on the surface of VEGF modified NiTi alloy were compared with nano-coated NiTi alloy and unmodified NiTi alloy. It has higher activity (p0.05 or p0.01), and the number of endothelial cells on the surface of VEGF modified NiTi alloy is more and the spreading area is larger. At the same time, the secretion of PGI2 and NO on the surface of VEGF modified NiTi alloy was significantly higher than that of unmodified NiTi alloy (p0.05). This study has accumulated useful experimental data for the wide application of NiTi alloy in biomedical field.
【学位授予单位】：重庆大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.08
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