骨内植入物表面酶响应型涂层改性精准防治术后相关感染研究  

Enzyme-responsive coating modification on surface of intraosseous implants for precise prevention and treatment of postoperative related infection

作　　者：余海平[1] 黄家琦 李哲 肖捷[1] 白欣[1] 汪燕萍 廖鑫 YU Haiping;HUANG Jiaqi;LI Zhe;XIAO Jie;BAI Xin;WANG Yanping;LIAO Xin(Department of Orthopedics,Jiande Municipal First People’s Hospital,Jiande,Zhejiang 311600,China)

机构地区：[1]建德市第一人民医院骨科,浙江建德311600

出　　处：《重庆医学》2023年第22期3397-3404,共8页Chongqing medicine

基　　金：浙江省医药卫生科技计划项目(2022RC244);浙江省杭州市医药卫生科技项目(B20220091)。

摘　　要：目的在骨内植入物表面构建10个双层蒙脱土/透明质酸-替考兰宁[(MMT/HA-TEC)10]多层膜抗菌涂层,探索其酶响应性抗菌效果。方法通过原子粒显微镜观察多层膜抗菌涂层表面形貌,光谱椭圆偏振法测试其厚度和全自动接触角测量仪测量其表面接触角。采用抑菌环实验探索抗菌涂层的抗生素在接触酶和细菌的情况下的缓释规律。电导率、细胞外蛋白水平检测和抑菌效果测试,宏观展示抗菌涂层的抗菌效果;采用高压蒸汽灭菌法测试抗菌涂层的抑菌稳定性。采用细菌活死染色实验探索抗菌涂层表面清除死亡细菌的能力。采用CCK-8实验法分析抗菌涂层的生物相容性。动物实验检测抗菌涂层的体内抗菌效果。结果(MMT/HA-TEC)10多层膜抗菌涂层在组装过程中,厚度呈线性增长,从(8.16±1.36)nm增长至(113.28±4.44)nm。接触角也明显变大,从组装前的27.1°变为组装后的65.0°。(MMT/HA-TEC)10多层膜抗菌涂层在与金黄色葡萄球菌和透明质酸酶(HAS)溶液孵育后逐渐降解,并显示出良好的浓度依赖性降解特性。抗菌涂层是通过改变细菌细胞膜的通透性来实现高效的杀菌效果,在经过高压蒸汽灭菌处理后其杀菌率依然>99%,并且在一定程度上可以清除材料表面的死亡细菌。CCK-8实验显示(MMT/HA-TEC)10多层膜抗菌涂层具有良好的生物相容性。动物实验表明(MMT/HA-TEC)10多层膜抗菌涂层在体内依然具有良好的抗菌作用。结论(MMT/HA-TEC)10多层膜抗菌涂层具有高效的酶响应性抗菌功能。Objective To construct the(MMT/HA-TEC)10 multilayer antibacterial coating on the surface of intraosseous implants,and to explore its enzyme-responsive antibacterial effect.Methods The antibacterial coating structure was observed by the atomic particle microscopy,its thickness was measured by the spectral ellipsometry method and the surface contact angle was measured by the fully automatic contact angle measurement instrument.The inhibition loop experiment was used to investigate the slow release rule of antibiotics with antibacterial coating in contact with enzymes and bacteria.The conductivity rate,extracellular protein level detection and bacteriostatic effect test macroscopically exhibited the bacteriostatic effect of the antibacterial coating,and the antibacterial stability of the coating was tested by the high pressure steam sterilization.The bacterial live-dead staining experiments was used to explore the ability of antibacterial coating surface for clearing the dead bacteria.The CCK-8 assay was used to analyze the biocompatibility of antibacterial coating.The animal experiment was used to emphatically explore the in vivo antibacterial effect of the antibacterial coating.Results The(MMT/HA-TEC)10 antibacterial coating thickness showed the linear growth from(8.16±1.36)nm to(113.28±4.44)nm during its assembly process.The contact angle was also significantly enlarged,which was changed from 27.1°before assembly to 65.0°after assembly.The(MMT/HA-TEC)10 antibacterial coating was gradually degraded after the incubation with Staphylococcus aureus and hyaluronidase(HAS)solutions,showing the good concentration dependent degradation properties.The antibacterial coating achieved the efficient bactericidal effect by changing the permeability of bacterial cell membrane.After the high pressure steam sterilization treatment,its bactericidal rate was still greater than 99%,and it could remove the dead bacteria on the material surface to some extent.The CCK-8 test showed that the(MMT/HA-TEC)10 antibacterial coating posse

关 键 词：抗菌 酶响应性 抑菌环 电导率 活死染色 

分 类 号：R318.08[医药卫生—生物医学工程]