超声喷涂技术制备细胞膜涂层工艺初探  

Erythrocyte Membrane Coating Technology by Ultrasonic Spraying

作　　者：吉诚 安永琪 王雅楠[2] 蒋青[2,3] 罗日方 王云兵 JI Cheng;AN Yongqi;WANG Ya'nan;JIANG Qing;LUO Rifang;WANG Yunbing(College of Materials Science and Engineering,Sichuan University,Chengdu 610065,China;College of Biomedical Engineering,Sichuan University,Chengdu 610065,China;National Engineering Research Center for Biomaterials,Chengdu 610065,China)

机构地区：[1]四川大学材料科学与工程学院,成都610065 [2]四川大学生物医学工程学院,成都610065 [3]国家生物医学材料工程技术研究中心,成都610065

出　　处：《表面技术》2024年第23期78-87,共10页Surface Technology

基　　金：国家自然科学基金(32371401);四川大学青年科技学术带头人培育项目(2023-08)。

摘　　要：目的利用超声喷涂技术在宏观界面构建具有抗污、抗凝血功能的细胞膜涂层,以提高涂层制备过程的原料利用率、涂层的均匀性和稳定性。方法使用超声喷涂技术将制备的红细胞膜(EMs)逐层喷涂在基底界面,使用荧光显微镜、扫描电子显微镜、水接触角等验证红细胞膜涂层(EMC)的组装过程、涂层稳定性的影响因素和涂层在多种材料表面的稳定涂覆。并通过蛋白吸附实验、血小板黏附和半体内血液循环实验评价涂层的抗污性能和血液相容性。结果在膜蛋白质量浓度为1 mg/mL的条件下,约30μL/cm^(2)的喷涂量,可获得厚度约500 nm的红细胞膜涂层,并且随着红细胞膜喷涂量的增加,红细胞膜涂层厚度逐渐增加至1~2μm。由于超声喷涂时涉及液滴快速蒸发和膜成分融合的过程,分散介质的盐离子浓度以及基底界面的特性将影响涂层的稳定性。超纯水体系有助于避免盐结晶行为对膜融合的阻碍。加热基底有助于加快喷涂速率,抑制不均匀的咖啡环结构。此外,富含官能团的亲水界面可以有效促进微液滴的融合和铺展,提高涂层的稳定性。优化工艺后,利用超声喷涂技术可在无机、金属和多种聚合物基材表面形成稳定的红细胞膜涂层,在基底表面形成水接触角34°的亲水性涂层。蛋白吸附实验表明,制得的红细胞膜涂层具有良好的抗污性能。血液相容性实验表明,红细胞膜涂层可有效减少血小板的黏附和激活以及材料表面血栓形成。结论超声喷涂技术可在多种材料表面形成可控、均匀和稳定的红细胞膜涂层,赋予材料表面良好的抗污、抗凝血性能。Macroscopic cell membrane coatings can successfully preserve the functional properties of native cell membranes,endowing the materials with good anti-fouling properties,immune evasion,and enhanced biocompatibility.However,current coating technologies,such as drop coating and dip coating,exhibit the shortcoming of high cell membrane·consumption,prolonged preparation time,and poor coating stability.Meanwhile,surface modification methods such as superhydrophilic interfaces,poly(tannic acid)coatings and click-chemical reaction interfaces have been successfully implemented to construct stable macroscopic cell membrane coatings,but the procedure of substrate pretreatment is relatively complicated.Ultrasonic spraying technology can be used to efficiently deposit atomized microdroplets to form a uniform coating.In this study,erythrocyte membranes(EMs)were investigated as a model for the application of ultrasonic spraying technology to construct cell membrane coatings at the macroscopic scale.The ultrasonic spraying technology significantly reduced the consumption of cell membranes,requiring only about 0.03 mg of membrane proteins to completely cover 1 cm^(2) of substrate surface,forming an erythrocyte membrane coating(EMC)of~560 nm thickness.As the spray volume increased,the atomized EMs were deposited on the substrate surface and fused together.Due to the controllable properties of the ultrasonic spraying technology,the deposited weight and coating thickness of EMCs tended to increase linearly.The dispersing medium of the EMs,the temperature and the interfacial properties of the substrate showed a great effect on the assembly of EMs and the stability of the coating.Scanning electron microscopy revealed that the PBS solution and the ultrapure water system resulted in a distinct morphology of EMCs.The ultrapure water system avoided the obstruction of membrane assembly by salt crystallization,facilitating the formation of a continuous coating on the substrate and preventing coating loss after rinsing.In addition,the eleva

关 键 词：超声喷涂 细胞膜涂层 红细胞膜 抗污界面 血液相容性 表面改性 

分 类 号：R318[医药卫生—生物医学工程] TG174.442[医药卫生—基础医学]