层层自组装法载药型人工晶状体的制备及其性能研究

发布时间：2018-07-11 13:37

本文选题：静电层层自组装 + 人工晶状体　；参考：《浙江大学》2016年博士论文

【摘要】：目的：利用静电层层自组装法在人工晶状体上修饰指数增长型载药多层膜,并探索改性后的人工晶状体其理化性质、光学及力学性质、体外细胞毒性以及杀菌性能。方法：利用次大气等离子体技术结合静电层层自组装技术,以妥布霉素和阿米卡星作为装载药物,在人工晶状体表面组装聚乙酰亚胺-(透明质酸-聚赖氨酸/妥布霉素或阿米卡星)。-透明质酸多层膜。利用接触角测量检测其亲疏水性能。利用石英晶体微天平(QCM)全程模拟组装过程,检测自组装膜的质量、厚度及粘弹性能随层数增加的变化,并验证指数模式增长。通过场发射扫描电镜和原子力显微镜对改性后的人工晶状体进行表面形貌检测,并计算表面粗糙度。利用X-射线光电子能谱对改性后的人工晶状体进行表面元素构成分析,验证组装过程的成功性。应用SFDA关于IOL质量检测的国家标准(YY-0290)对改性后IOL进行相关测试,包括像质、光焦度、光谱透过率和动态疲劳耐久性等。将改性后人工晶状体浸泡的细胞培养液与人晶状体上皮细胞及人视网膜色素上皮细胞共同培养,并用CCK-8实验检测对这种两种细胞的毒性作用。将改性后的人工晶状体与表皮葡萄球菌共同培养,通过荧光染料试剂盒在荧光显微镜下观察活细菌与死细菌数量,检测IOL的体外杀菌效果。结果：通过接触角测定,改性后的IOL亲水性大幅度提高。通过QCM模拟组装过程,证实了组装的成功性,且通过改进组装体系条件,IOL表面物质的质量与厚度随层数增加呈指数增长模式,大大提高了改性效率和载药量。场发射扫描电镜和原子力显微镜显示改性后IOL表面形貌和粗糙度无明显改变。XPS显示表面元素成分变化是组装成功的表现。对改性后IOL进行光焦度、像质、光谱透过率和动态疲劳耐久性测试后发现,新型IOL各个方面均符合国家标准(YY-0290)。CCK-8实验证明,改性后人工晶状体对人晶状体上皮细胞和人视网膜色素上皮无明显毒性作用。体外杀菌实验证明,载药型IOL可有效杀死表皮葡萄球菌。结论：通过静电层层自组装法成功制备出表面(透明质酸／妥布霉素(或阿米卡星)-聚赖氨酸)。多层膜修饰的新型人工晶状体,该多层膜的质量、厚度可随层数增加实现指数增长模式。所使用的该种改性方法不仅提高了IOL表面亲水性,且不影响表面形貌及粗糙度,改性后IOL光学、力学性能符合国家标准(YY-0290),对于体外人晶状体上皮细胞和人视网膜色素上皮细胞无明显毒性作用,在体外实验中证明具备杀灭表皮葡萄球菌的效果。这种新型载药型人工晶状体有望用于防治白内障术后眼内炎的发生。
[Abstract]:Objective: to study the physical and chemical properties, optical and mechanical properties, cytotoxicity and bactericidal properties of the modified intraocular lens (IOL) by electrostatic layer self-assembly method. Methods: tobramycin and amikacin were used as loading drugs by using subatmospheric plasma technique and electrostatic layer self-assembly technique. The polyimide- (hyaluronic acid-polylysine / tobramycin or amikacin) was assembled on the intraocular lens surface. Its hydrophilicity was measured by contact angle measurement. Quartz crystal microbalance (QCM) was used to simulate the whole assembly process. The mass, thickness and viscoelastic energy of the self-assembled films were measured with the increase of the number of layers, and the exponential mode growth was verified. The surface morphology of the modified intraocular lens was measured by field emission scanning electron microscope (SEM) and atomic force microscope (AFM), and the surface roughness was calculated. The surface element composition of the modified IOL was analyzed by X- ray photoelectron spectroscopy (XPS) to verify the success of the assembly process. The modified IOL was tested by SFDA (YY-0290) national standard for IOL quality, including image quality, light focus, spectral transmittance and dynamic fatigue durability. The culture medium of modified intraocular lens was co-cultured with human lens epithelial cells and human retinal pigment epithelial cells. The toxicity of these two cells was detected by CCK-8 experiment. The modified intraocular lens was co-cultured with Staphylococcus epidermidis. The quantity of live and dead bacteria was observed by fluorescent dye kit and the bactericidal effect of IOL was detected in vitro. Results: the hydrophilicity of modified IOL was greatly improved by measuring contact angle. By simulating the assembly process by QCM, the success of the assembly was proved, and the quality and thickness of the surface matter of IOL increased exponentially with the increase of the number of layers by improving the conditions of the assembly system, which greatly improved the modification efficiency and drug loading. Field emission scanning electron microscopy (SEM) and atomic force microscope (AFM) showed that the surface morphology and roughness of the modified IOL showed no obvious change. XPS showed that the change of surface element composition was the result of successful assembly. It was found that all aspects of the modified IOL were in accordance with the national standard (YY-0290). CCK-8 experiment proved that the light coke, image quality, spectral transmittance and dynamic fatigue durability of the modified IOL were all in line with the national standard (YY-0290). The modified intraocular lens has no toxic effect on human lens epithelial cells and retinal pigment epithelium. In vitro bactericidal test showed that drug-loaded IOL could effectively kill Staphylococcus epidermidis. Conclusion: the surface (hyaluronic acid / tobramycin (or amikacin)-polylysine) was successfully prepared by electrostatic layer self-assembly. The mass and thickness of the multilayer IOL can increase exponentially with the increase of the number of layers. The modified method not only improves the surface hydrophilicity of IOL, but also does not affect the surface morphology and roughness. The mechanical properties were in accordance with the national standard (YY-0290) and had no obvious toxicity to human lens epithelial cells and human retinal pigment epithelial cells in vitro. It was proved in vitro that it had the effect of killing Staphylococcus epidermidis. This new drug-loaded intraocular lens is expected to be used to prevent and treat endophthalmitis after cataract surgery.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R779.6;R318.1

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