吉诚,安永琪,王雅楠,蒋青,罗日方,王云兵.超声喷涂技术制备细胞膜涂层工艺初探[J].表面技术,2024,53(23):78-87.
JI Cheng,AN Yongqi,WANG Ya'nan,JIANG Qing,LUO Rifang,WANG Yunbing.Erythrocyte Membrane Coating Technology by Ultrasonic Spraying[J].Surface Technology,2024,53(23):78-87 |
| 超声喷涂技术制备细胞膜涂层工艺初探 |
| Erythrocyte Membrane Coating Technology by Ultrasonic Spraying |
| 投稿时间：2024-09-06 修订日期：2024-11-06 |
| DOI：10.16490/j.cnki.issn.1001-3660.2024.23.006 |
| 中文关键词: 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 |
| 英文关键词:ultrasonic spraying cell membrane coating erythrocyte membrane antifouling interface blood compatibility surface modification |
| 基金项目: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) |
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| Author | Institution |
| JI Cheng | College of Materials Science and Engineering, Chengdu 610065, China |
| AN Yongqi | College of Biomedical Engineering, Sichuan University, Chengdu 610065, China |
| WANG Ya'nan | College of Biomedical Engineering, Sichuan University, Chengdu 610065, China |
| JIANG Qing | College of Biomedical Engineering, Sichuan University, Chengdu 610065, China;National Engineering Research Center for Biomaterials, Chengdu 610065, China |
| LUO Rifang | College of Biomedical Engineering, Sichuan University, Chengdu 610065, China;National Engineering Research Center for Biomaterials, Chengdu 610065, China |
| WANG Yunbing | College of Biomedical Engineering, Sichuan University, Chengdu 610065, China;National Engineering Research Center for Biomaterials, Chengdu 610065, China |
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| 中文摘要: |
| 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30 μL/cm2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1~2 μm。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。 |
| 英文摘要: |
| 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 cm2 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 elevated substrate temperature accelerated the spraying rate and inhibited the inhomogeneous coffee ring structure. Compared to room temperature conditions, heating the substrate to 40 ℃ was beneficial in preventing localized agglomeration of microdroplets and improving coating uniformity during the spraying process. Furthermore, the hydrophilic interface enriched with functional groups could effectively promote the fusion and spreading of microdroplets and increase the stability of the coating. Fluorescence microscopy demonstrated superior stability of EMCs on oxygen plasma-treated and polydopamine-treated substrates in a flow environment in comparison to hydrophobic polydimethylsiloxane (PDMS). After optimization of the coating technology, ultrasonic spraying was utilized to form uniform and stable EMCs on a wide range of macroscopic materials. The prepared EMCs formed a hydrophilic coating with a water contact angle of 34°. Due to the hydrophilic phospholipid bilayer on the EMC surface, which exhibited a stable hydration layer and a natural negative charge, the prepared EMCs showed less protein adsorption and exhibited good antifouling properties compared to the bare stainless steel. In addition, the blood compatibility and ex vivo circulation experiments showed that the erythrocyte membrane coatings were capable of reducing platelet adhesion and activation, as well as thrombus formation. Due to their excellent antifouling and blood compatibility, EMCs have significant potential in the field of surface modification of biochips and blood contact materials. |
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