具有抗细菌黏附和杀菌双重功能的(HA/CHI-Van)5涂层的制备

目的：研究开发一种涂覆(HA/CHI-Van)5多层膜结构的骨科内植入物以用于预防骨内植入物感染。方法：将透明质酸（HA）和壳聚糖-万古霉素（CHI-Van）溶液进行层层自组装，制备得到(HA/CHI-Van)5多层膜结构；通过zeta电位检测组装过程，通过活细菌染色实验评价多层膜的抗细菌黏附效果，通过越狱实验和FE-SEM实验检测体外抗菌效果。SD大鼠随机分为空白组、(HA/CHI-Van)5组和假手术组，并以X线、Micro-CT、骨组织切片等方法检测该材料的体内动物实验效果。结果：每次组装不同的单层，zeta电位交替变化，证明多层膜组装成功。活细菌染色实验显示多层膜结构中的CHI具有一定的抗细菌黏附作用。越狱实验和FE-SEM实验证明(HA/CHI-Van)5多层膜结构在体外具有良好的抗菌作用。在体外实验中，(HA/CHI-Van)5组的X线评分为0.7，而空白组的评分为12.3，(HA/CHI-Van)5组在骨密度、骨小梁数量、骨体积分数、骨组织切片方面均优于空白组（均P＜0.05），且非常接近假手术组（均P＞0.05）。结论：通过在骨科内植入物表面涂覆(HA/CHI-Van)5多层膜结构能够获得一定的抗细菌黏附功能和良好的抗菌作用，可以预防骨内植入物感染。

Preparation of anti-infective and bactericidal dual-functional multilayer membrane structure

Objective: To study an implant coated with a (HA/CHI-Van)5 multilayer membrane structure to prevent implant infection. Methods: The HA and CHI-Van solution were assembled to prepare a (HA/CHI-Van)5 multilayer film structure, and the assembly process was detected by zeta potential. The antibacterial adhesion function of the multilayer film was evaluated by a live bacterial staining test. And in vitro antibacterial effect was also tested by jailbreak experiments and FE-SEM experiment. Most importantly, adequate animal experiments were designed besides X-ray, Micro-CT and bone tissue sections were to discuss in vivo experiments. Results: Each time a different monolayer was assembled, the zeta potential alternated, demonstrating the successful assembly of the multilayer film. Live bacteria staining experiments showed that CHI in the multilayer membrane structure had a certain anti-bacterial adhesion effect. The jailbreak experiment and the FE-SEM experiment proved that the (HA/CHI-Van)5 multilayer membrane structure had a good antibacterial effect in vitro. The X-ray score of (HA/CHI-Van)5 group was 0.7, while the score of blank group was 12.3. (HA/CHI-Van)5 group was superior to blank group in bone mineral density, trabecular bone number, relative bone volume, and bone tissue section, and was very close to sham operation group. Conclusion: By applying a (HA/CHI-Van)5 multilayer membrane structure in the implant surface, a certain anti-bacterial adhesion and good antibacterial effect can be obtained, and the implant infection can be prevented.